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| THE 5TH ANNUAL MEETING OF GP-TCM RESEARCH ASSOCIATION-CUM-SUMMIT ON COMPENDIUM OF MATERIA MEDICA AND INNOVATIVE DRUG DISCOVERY IN CHINESE MEDICINE |
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| Year : 2016 | Volume
: 2
| Issue : 2 | Page : 72-94 |
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The 5th Annual Meeting of GP-TCM Research Association-cum-Summit on Compendium of Materia Medica and innovative drug discovery in Chinese medicine: 9 – 10 August 2016 Hong Kong Baptist University (Shaw Campus), Hong Kong
| Date of Web Publication | 8-Sep-2020 |
Correspondence Address:
 Source of Support: None, Conflict of Interest: None
How to cite this article:
. The 5th Annual Meeting of GP-TCM Research Association-cum-Summit on Compendium of Materia Medica and innovative drug discovery in Chinese medicine: 9 – 10 August 2016 Hong Kong Baptist University (Shaw Campus), Hong Kong. World J Tradit Chin Med 2016;2:72-94 |
How to cite this URL:
. The 5th Annual Meeting of GP-TCM Research Association-cum-Summit on Compendium of Materia Medica and innovative drug discovery in Chinese medicine: 9 – 10 August 2016 Hong Kong Baptist University (Shaw Campus), Hong Kong. World J Tradit Chin Med [serial online] 2016 [cited 2023 Dec 1];2:72-94. Available from: https://www.wjtcm.net/text.asp?2016/2/2/72/294528 |
Chinese medicine is one of the treasures of Chinese culture. In recent decades, its use has spread far and wide and there is increasing recognition of its value worldwide. The Meeting-cum-Summit provides an interactive platform for the exchange of the latest research findings of meeting participants to foster academic excellence in Chinese medicine.
Objectives
- Promote discussion and implementation of good practice in TCM research and development, high-quality evidence-based research on TCM as well as the integration of Chinese medicine and conventional medicine;
- Revisit the significant contributions made by Li Shizhen, the author of the Compendium of Materia Medica, to the development of Chinese medicine and discuss new insights into his work;
- Explore new strategies for and approaches to new drug discovery and the standardisation of Chinese materia medica with a view to contributing to the advancement, modernisation and internationalisation of Chinese medicine as well as health and well-being.
Themes
- Quality control
- Pharmacology and toxicology
- Regulatory matters and standardisation
- Clinical studies
- Acupuncture
- Compendium of Materia Medica
- New drug discovery strategies and approaches
The Meeting-cum-Summit is jointly organised by the School of Chinese Medicine of Hong Kong Baptist University, GP-TCM Research Association and TCM Pharmaceutical Analysis Specialty Committee of World Federation of Chinese Medicine Societies.a
| Organising Committee | |  |
Prof. De-An Guo (Chairman)
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, China
Prof. Rudolf Bauer
Institute of Pharmaceutical Sciences, Karl-Franzens-University Graz, Austria
Prof. Zhao-Xiang Bian
School of Chinese Medicine, Hong Kong Baptist University, Hong Kong
Prof. Kelvin Chan
Western Sydney University & University of Technology Sydney, Australia; Liverpool John Moores University, UK
Prof. Hen-Hong Chang
School of Chinese Medicine, China Medical University, Taiwan
Prof. Yuan-Shiun Chang
College of Pharmacy, China Medical University, Taiwan
Prof. Yung-Hsien Chang
China Medical University and Hospital, Taiwan
Prof. Shi-Lin Chen
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, China
Prof. Wan-Sheng Chen
Second Military Medical University, China
Prof. Pierre Duez
Department of Therapeutical Chemistry and Pharmacognosy, University of Mons, Belgium
Prof. Thomas Efferth
Department of Pharmaceutical Biology, Johannes Gutenberg University, Germany
Dr. Tai-Ping Fan
Faculty of Biology, University of Cambridge, UK
Prof. Jing-Yan Han
Health Science Centre, Peking University, China
Prof. Jin-Da Hao
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, China
Prof. Peter Hylands
Institute of Pharmaceutical Science, King’s College London, UK
Prof. Ling-Yi Kong
China Pharmaceutical University, China
Prof. Hai-Xue Kuang
Heilongjiang University of Chinese Medicine, China
Prof. Min Li
South China Normal University, China
Prof. Jian-Ping Liu
School of Basic Medical Sciences, Beijing University of Chinese Medicine, China
Prof. Ai-Ping Lu
School of Chinese Medicine, Hong Kong Baptist University, Hong Kong
Dr. Fan Qu
School of Medicine, Zhejiang University, China
Prof. Nicola Robinson
School of Health and Social Care, London South Bank University, UK
Prof. Monique Simmonds
The Royal Botanic Gardens, Kew, UK
Prof. Robert Verpoorte
Natural Products Laboratory, Leiden University, The Netherlands
Prof. Jia-Kui Wang
Chengdu University of Traditional Chinese Medicine, China
Prof. Xi-Jun Wang
Heilongjiang University of Chinese Medicine, China
Prof. Yi-Tao Wang
Institute of Chinese Medical Sciences, University of Macau, Macau
Prof. Vivian Wong
School of Chinese Medicine, The University of Hong Kong, Hong Kong
Prof. Hong-Xi Xu
School of Pharmacy, Shanghai University of Traditional Chinese Medicine, China
Dr. Qi-He Xu
Department of Renal Medicine, King’s College London, UK
Prof. Wei-Dong Zhang
Second Military Medical University, China
Prof. Zhi-Bin Zhang
Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, China
Dr. Zhi-Jie Zhang
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, China
Prof. Zhong-Zhen Zhao
School of Chinese Medicine, Hong Kong Baptist University, Hong Kong
Prof. Jin-Sheng Zheng
Institute for History of Chinese Medicine and Medical Literature, China Academy of Chinese Medical Sciences, China
| Oral Presentations | | |
| 0-01: A New Method For Quantitative the Glycoprotein in Biological Samples | | |
Yang Gao, Hongyue Li, Duoduo Xu, Mingxing Wang, Qipin Gao
Changchun University of Chinese Medicine, Key Laboratory of effective components of TCM, Ministry of Education, China. Changchun, 130117 China
Jilin Key Laboratory of Macromolecules in Chinese Drug. Changchun, 130117 China
Correspondence: Qipin Gao, e-mail: [email protected]
Objective: To set up a new method to detect the sugar complex in biological samples. Methods: The glycoprotein purified from the mycelium extract of Tremella fuciformis was marked with iodine through the iodine substitution reaction and the content of iodine, which is representative the amount of the marked trmella glycoprotein (ITG), was detected with ICP-MS. The methodology was performed and found the method is stable, sensitive, and accurate at the detecting the content of iodine substituted glycoprotein by ICP-MS. The method was used in the quantitative analysis of biological samples about their content of ITG, including blood and organs. Different biological samples were collected from rats after administration ITG orally and were tested their iodine contents by ICP-MS to calculating the amount of ITG in the samples. Result: The results suggested that the iodine glycoprotein in blood or organs is able to be detected by ICP-MS with sensitive, stable and accurate characters. Conclusion: Glycoprotein is easy to be labeled with iodine to form stable derivative in animal body and the iodine derivative of glycoprotein can be detected with ICP-M. The methodology studies reveal the method is sensitive, stable and easy to perform, which could be used wildly in the study on glycoprotein, speciously in its pharmacokinetic.
| O-02: Traditional Medicine, Natural Products, MMPS and the Molecular Basis of Therapeutics | | |
Bipin G. Nair
Amrita School of Biotechnology, Amrita Vishwa Vidyapeetham, Amrita University, Kollam, Kerala 690525, India
Correspondence: Bipin Nair, e-mail: [email protected]
Objective: To decipher the molecular mechanisms involved in the regulation of MMP-2 and MMP-9 by anacardic acid, isolated from cashew nut shell liquid and a natural product-like flavonoid analog, (I-3, II-3)-biacacetin and implications for cancer therapy. Methods: Anacardic acid from cashew shells and (I-3, II-3)-biacacetin synthesized through oxidative dimerization of poly-substituted diaryl-1,3-diketones with cerium ammonium nitrate followed by double cyclodehydration were characterized using IR, UV, HPLC, Mass Spectrometry and NMR. Human fibrosarcoma cells were treated with the compounds and the effect on gelatinase activity was studied using gelatin zymography, gelatin degradation and a fluorescence-based assay. qPCR analysis and Western blotting was used for expression studies. Results: Anacardic acid and (I-3, II-3)-biacacetin inhibits MMP-2 and MMP-9 gelatinolytic activity (1, 2). Furthermore, fluorescence-based studies with catalytic domain revealed that anacardic acid inhibits MMP-2 catalytic activity in a dose-dependent manner. The carboxylate group, hydroxyl group binds and the lipophilic C15 of anacardic acid were found to be essential for the inhibitory activity. Unlike the zinc dependent inhibition of gelatinases by anacardic acid, the inhibition by biacacetin is independent of zinc. Further, a combination of anacardic acid with biacacetin at concentrations that are 10 fold lower than that of either compound alone, resulted in significant enhancement of inhibitory activity. Conclusions: Anacardic acid and (I-3, II-3)-biacacetin act as potent inhibitors of gelatinase activity which in turn gives a strong impetus for the natural products drug discovery paradigm by providing a new template for MMP-2/MMP-9 drug discovery.
| O-03: Development of a Novel Phytopharmaceutical Medicine Derived from Traditional Chinese Medicine for the Treatment of Parkinson’S Disease | | |
Friedemann T a, Li M b, Fei Jc, Ying Y c, Wang WG c, Song J b, Kramer ER d, Schumacher U e, Leung AKM b, Schroeder S a
a HanseMerkur Center for Traditional Chinese Medicine at the University Medical Center Eppendorf, Hamburg
b Hong Kong Baptist-University, Hong Kong
cTongji University, Shanghai
d Centre for Molecular Neurobiology Hamburg (ZMNH), Hamburg
e University Medical Center Hamburg-Eppendorf, Department of Anatomy 2, Hamburg
Correspondence: Sven Schroeder, e-mail: [email protected]
Introduction: Alzheimer's disease (AD) and Parkinson's disease (PD) are the third most common cause of death in industrialized countries. Given the lack of neuroprotective conventional medicines, investigation of Chinese Medicine herbs (CMH) holds a high potential for the development of novel therapeutic concepts. Materials and Methods: We established a hypothesis driven screening approach to identify potentially neuroprotective CMH [1]. The initial screening of watery extracts for their neuroprotective effect against oxidative stress induced cellular damages revealed that Coptis chinensis (CC) was the most promising herb. Results: Further analysis showed that pre-treatment of neuroblastoma cells with CC or Coptisine has three effects: attenuation of oxidative stress-induced decrease in cell viability, protection of the mitochondrial membrane potential, and reduction of apoptosis. PD related investigations revealed that CC, Coptisine and Berberine protect neuroblastoma cells and mice from MPP+/MPTP induced toxicity, respectively, in a dose dependent manner. These findings suggest that neuroprotective effects of CC might be caused by an improvement of mitochondrial function, stabilization of intracellular ATP concentration, attenuation of mitochondrial permeability transition, reduction of cytochrome C release and prevention of apoptosis. Conclusions: These results can be seen as a promising starting point for the development of new treatment concepts for PD and other neurodegenerative diseases. However, multi-factorial diseases like PD require a multi-target approach for affecting different dysregulated pathways. Hence, we aim to combine several herbs (or compounds) which act via different pathways to achieve additive or synergistic effects for an increasement of the therapeutic efficacy.
| 0-04: Indirect Genotoxicity of Herbal Medicines | | |
Amandine Nachtergael, Mélanie Poivre, Pierre Duez
Unit of Therapeutic Chemistry and Pharmacognosy, Faculty of Medicine and Pharmacy, Research Institute for Health Sciences and Technology, University of Mons (UMONS), 20 Place du Parc, 7000 Mons (Belgium)
Correspondence: Pierre Duez, e-mail: [email protected]
Herbal medicines are widely used worldwide, either as primary health care or complementary medicines. The toxicology of most herbs has however not been thoroughly investigated and their safety is generally assumed from long-standing use. For mid- to long-term toxicities, such as genotoxicity, that are generally not detected through traditional use, the situation is more complex and requires more scientific evidence for their putative harmlessness. To this end, a plethora of methods [1] exist, more or less satisfying; structural alerts, in silico and classical in vitro and in vivo predictive methods are often used on a herb-by-herb basis. These tools are still a cornerstone for toxicological evaluation but appear ill-suited to study multicomponent and variable herbal mixtures [2]. The European Committee on Herbal Medicinal Products (HMPC) has proposed a genotoxicity testing guideline based on the Ames test; this strategy is however heavily debated as some ubiquitous compounds, Ames-positive but not genotoxic to eukaryotes, could mask hazardous genotoxins.
Also many factors can compromise the safety of herbal treatments, ranging from intrinsic factors of quality to external contaminations or various pharmacological/pharmacokinetic interactions. Of note, we recently provided evidence for indirect genotoxicity, i.e. induction or potentiation of a herb genotoxicity by a co-administered herb (Aristolochia-Magnolia) [3]. Given the various mechanisms for genome maintenance, the possibilities of such interactions are not negligible and should be considered in developing further guidelines to test the actual preparations/mixtures delivered to patients.
| O-05: On a Nomenclature Appropriate to Orientation for Chinese Medicines | | |
Bo-Ying Ma, Alicia Grant
Xinglin Postgraduate College of TCM
Correspondence: Bo-Ying Ma, e-mail: [email protected]
Chinese medicine is a great treasure house, which safeguarded and has continually been safeguarding the health and well-being, prevention and treatment of diseases of people in China and worldwide. When western medicine transferred into China ca. 200 years ago, there were not many arguments on the orientation of Chinese medicine but during the recent 20 years this question has arisen and it is creating many obscurities and puzzles for western people since Chinese medicine became popular and widely used. What is an appropriate nomenclature to the orientation of Chinese medicines? In the west, for the taking or oral ingestion there are only two categories: FOOD or MEDICINES in which the orientation of Chinese medicines was omitted. In the tradition of Chinese culture medicines and food come from the same source and they could be used for both food and medicine according to their purpose and who gave them. However, western administrative management does not agree to mix up sale and use in the market. There are two different systems of medicine in the world: western medicine and Chinese medicine that, in this case, cannot get along with each other. Chinese medicines cannot accept the standard of western medicine because they do not belong to the western scope of medicine. The orientation of Chinese medicines should be placed between food and medicines in the western categories. We believe that this objective fact should now be acknowledged and respected. This paper would give reasonable evidence from history, traditional theory and clinical practice of Chinese medicine for the suggestion.
| O-07: Comparison of Enrich-Blood Bioactivity Among Various Species and Grade of Danggui Herbs | | |
Bowen Yang a, Xi Li b, Xiaoxiao Wang c, Qinwen Yang a, Zuyuan Rong b, Lihong Zhang b, Shuang He b, Hongping Wei b, Guanghua Lu a
a School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
b Sichuan Institute for Food and Drug Control, Chengdu 611730, China
c Deyang Institute for Food and Drug Control, Deyang 618000, China;
Correspondence: Guanghua Lu, e-mail: [email protected]; [email protected]
Objective: To compare the enrich-blood bioactivity (EBB) among different grades of Chinese Danggui (CDG) and related herbs for quality assessment. Methods: 7 CDG samples were collected from various cultivating areas in China and divided into five grades by the weight of single root, i.e. first (70-50 g), second (40-25 g), third (20-10 g), fourth (10-5 g) and fifth-grade (5-1 g). Meanwhile, 47 herbal samples including Japanese Danggui (JDG), Korean Danggui (KDG), Lovage root (LR) and Angelica archangelica root (AAR) were collected from different countries. These samples were extracted in ethanol and water, respectively, and then quantified the EBBs by animal experiment. Red blood cell was chosen as maker. Cyclosporine A was reference. The variance analysis and correlation analysis were further applied to assess these samples. Results: EBB in CDG samples were 33.789 U (n = 2, first-grade), 25.976 U (n = 6, second-grade), 23.900 U (n = 5, third-grade), 21.595 U (n = 5, fourth-grade) and 15.381 U (n = 4, fifth-grade). Meanwhile, The EBB in CDG, KDG, LR and JDG were estimated to be 27.976 U (n = 9), 15.334 U (n = 4), 14.897 U (n = 4) and 7.664 U (n = 5), respectively; whilst three AAR samples were not found this bioactivity. The differences were significant among grades of CDG samples and species samples. Conclusion: The larger CDG is better, which is in consistence with the results of experience identification. The efficacy of various Danggui species are different and cannot be mixed and substituted in clinical uses.
Acknowledgements: National Basic Science Personnel Training Fund (No. J1310034-02).
| Posters Presentations | | |
| P-01: Standardization on Clinical Pharmacy Terminology of Chinese Medicine for Development of Electronic Health Record (EHR) Sharing System in Hong Kong | | |
Dawn Au, Vincent Kan, Winnie Szeto, Timothy Yung
Chief Pharmacist’s Office, Hong Kong Hospital Authority, 10/F, Block A, Multicentre, PYNEH, Chai Wan, Hong Kong
Correspondence: Dawn Au, e-mail: [email protected]
Introduction: An electronic Health Record (eHR) Sharing System is developed to serve as a platform for health record sharing between public and private healthcare organizations with patients’ consent. Standardization of terminology in the health record is the pre-requisite of communication among different users in the eHR to ensure clinical efficacy and safety. As for Chinese medicine (CM), standardization on clinical pharmacy terminology is crucial since the use of a synonym of herbs could result in potential life-threatening events. Therefore, it is essential to develop a standard CM terminology database to ensure accurate information with the same understanding which is delivered and interpreted by all users, including clinical professions, health service organizations and Government Departments, in order to avoid miscommunication, misinterpretation and prevent medication errors. Methods: “Terminology-principles-and-methods” was established to produce CM pharmacy standards through literature review, herbal sample collection for special cases and discussion among various experts in the specific eHR Information Standards Domain Group on Chinese Medicine Pharmacy Terminology. Results: The initial design and architecture of the Chinese Herbal Medicine Vocabulary Table (CHMVT) has been established. Names (Chinese and Latin names) and sources of origin of raw herbs, decoction pieces (Yin Pian), as well as prescribing and dispensing terminology, including special decoction method, processing method and units, were standardized and incorporated into the CHMVT. In addition, sustainable principles agreed by the domain group were compiled for terminology management. Conclusion: Standardization method of CM pharmacy terminology and its corresponding editorial principle have been developed for the sake of safety, reliability, efficiency, interchangeability, and to facilitate communication with electronic CM data in the future. It is a Hong Kong based terminology database which can be applied to international model for standardization on Chinese medicine. The data management, storage and exchanges, as well as the relationship of “concepts” development and creation, will be needed to enhance data accuracy and quality for health information exchange in the future. Therefore, the professional work on standardizing the CM terminology is a long lasting mission and requires the concerted effort of multi-disciplines and stakeholders.
| P-02: Protective Effect of Sanggua Drink on Pancreatic Islet in type 2 Diabetes Mellitus Rats Induced by Streptozotocin | | |
Yu Cai a,b, Ying Wang a,b, Fei Zhi a,b, Qi-Chang Xing a,b and Yun-Zhong Chen a,b
a College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, PRC
b Institute of Engineering Technology of Chinese Traditional Medicine and Health Food of Hubei Province, Wuhan 430065, PRC
Correspondence: Yun-Zhong Chen, email: [email protected]
Introduction: Sanggua Drink (SGD) consists of Momordica charantia, Mulberry leaves, Pueraria lobata and Dioscorea opposite which have been extensively prescribed due to notable medicinal benefits. This article aims to investigate the protective effects of SGD on pancreatic islet in type 2 diabetic mellitus rats. Materials and Methods: Streptozotocin-induced diabetic model of rats were divided into four groups: normal control group, model group, positive contorl group and SGD water extraction group. The rats were oral administered once a day, continuously for 42 days. The levels of blood glucose, SOD, MDA, CAT and GSH-PX were determined. The technology of paraffin wax slice was used to make the pathological section of pancreas and the pathological change of pancreas was evaluated by hematoxylin/eosin staining of insulin. Results: Compared with normal control group, the level of MDA was increased significantly in model group, whereas the contents of SOD, CAT and GSH-PX were decreased significantly (P<0.01 or P<0.05). Compared with model group, the level of MDA was significantly decreased in positive group and treated group, and the contents of SOD, CAT and GSH-PX were significantly increased (P<0.01 or P<0.05). At the same time , SGD treated T2DM showed significantly ameliorated pathological changes and infIammation reduction in pancreas observed under light microscope. Conclusions: It is suggested that SGD could have the protective effect on the STZ-induced injury of pancreatic islet in T2DM rats through increasing antioxidant enzyme activity and reducing oxidative stress. The study provides the experimental basis for clinical application.
| P-03: Effects of Dioscorea Opposita Thunb. Polysaccharide on the Glycolipid Metabolism and Antioxidation in Type 2 Diabetic Rats | | |
Fei Zhi, Ying Wang, Qi-chang Xing, Yu Cai, Chun-chao Yan, Yun-zhong Chen
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China
Correspondence: Yun-zhong Chen , e-mail [email protected]
Objective: To investigate effects of the Dioscorea opposita Thunb. polysaccharide on the glycolipid metabolism and antioxidation in type 2 diabetic rats. Methods: SD rats were constructed by high-fat-diet and combined with intraperitoneal injection of streptozocin 40 mg• kg-1, whose blood glucose greater than 11.1mmol/L is successful diabetic mold. The model rats were randomly divided into diabetic model group, positive control group (metformin, 150 mg•kg-1), Dioscorea opposita Thunb. polysaccharide groups(DTP)(400 mg• kg -1) and the normal control group was also designed. After a 6-week treatment, the general condition of rats was observed. Body weight, as well as the blood glucose, the amount of hepatic glycogen, TG, TC, LDL-c, HDL-c, MDA, NO and the activtiy of T-SOD, CAT, GSH-PX were tested respectively. Results: Compared with the model group, DTP effetively relieved symptoms of type 2 dibetic rats, decreased the levels of the blood glucose, TC, and LDL-c , increased the amount of hepatic glycogen, enhanced the activtiy of CAT and GSH-PX. Conclusions: DTP shows a significantly protective effects on type 2 diabetic rats. The mechanism may be related to the reduction of hepatic glycogen decomposition or an increase synthesis of hepatic glycogen and the improvement of antioxidation.
| P-04: Studies on the Inhibition Effects to Bacterial and Bacterial-Biofilm of Ginsenosides | | |
Duoduo Xu a,b Qipin Gao a,b
a R & D Center, Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China
b Key Laboratory of Macromolecules from Traditional Chinese Medicine, Changchun, Jilin, 130117, China
Correspondence: Qipin Gao, e-mail: [email protected]
Objective: To establish a model of bacteria and bacteria-biofilm (BBF) of E. coli and S. aureus in vitro and to investigate the effect of ginsenosides on the models. Methods: BBF was cultured and formed in 96-well plates and then the internal morphology and structure of BBF was observed by SEM and LSCM at 6, 12, 18, 24 and 48 hours, respectively. The MTT method was used to investigate the inhibitory effect to bacterial and their BBF. Results: With the increase of culture time, the number of bacteria increased gradually. When they were cultured to 24 hours, BBF of both E. coli and S. aureus reached maturity. When the concentration was 1.0 mg/ml, The inhibition rates on planktonic bacteria of E.coli of ginsenosides Rb1, Rb2, Rb3, Rc, Rd, Re was 81.81% 81.28%, 85.08%, 84.67%, 85.89%, 25.12%, while the inhibition on E. coli BBF was 48.51%, 55.57%, 44.53%, 52.80% , 70.12%, 12.15%, respectively. The inhibition rate of the bacteria and BBF of S. aureus was 39.44%, 30.20%, 39.75%, 46.36%, 26.69%, and 48.22%, 31.19%, 50.72%, 61.83%, 11.22%, respectively. The inhibition rate and concentration showed a good quantity-effect relation. Conclusion: Ginsenoside Rb1, Rb2, Rb3, Rc, Rd have a certain inhibitory effect on bacteria and BBF of E. coli and S. aureus. Among them, Rd has the strongest effect, while Re has almost no anti-bacterial activity. Overall, the effect of Ginsenoside on E. coli is stronger than those of S. aureus.
| P-05: Preparation, Characterization and Cytokine-Stimulating Effect of the Oligosaccharides From Tremella Fuciformis Berk | | |
Yang Gao a,b, Duoduo Xu a,b, Mingxing Wang a, Yanqiu Jia a, Qipin Gao a,b
a R & D Center , Changchun University of Chinese medicine, Changchun, Jilin, 130117, China
b Key Laboratory of Macromolecules from Traditional Chinese Medicine, Changchun, Jilin, 130117, China
Correspondence: Qipin Gao, e-mail: [email protected]
Objective: Study on the cytokine-stimulating effect of the oligosaccharides from Tremella fuciformis Berk. Methods: 1. Hydrolysis of TP. 2.The Preparation of oligosaccharide broken from the main chain of TP. 3.Methylation Analysis. 4.Electro spraying ionization-mass spectrometry (ESI-MS). 5.Cytokine stimulating activity. Results: The tremella polysaccharide (TP) has significant cytokine-stimulating activity and was very big molecules. For studying its smallest active structure which shall be profitable in further utilization and research, TP was hydrolyzed and fractionated to obtain the fragments which was broken down from its (1→3)-mannan backbone. The fragments were isolated with gel-filtration and HPLC chromatography, three oligosaccharide fractions (L-4-1, 2, 3), which composed with di-, tri-saccharides; tetra-, penta-saccharides; and hex-, hepta-., octa-saccharide, respectively, were obtained. Methylation was performed to evaluate the glycosidic linkage in the oligo-mannan, and the result showed that the L-4-2 and 3 were linked mainly by a 1-3 glycosidic band, and approximately 3% or | 6% branching occurred in the 6 position of L-4-2 and L-4-3, respectively. The cytokine-stimulating experiment showed that the L-4-1 had no cytokine stimulating activity, L-4-2 had weak activity, and L-4-3 showed strong activity, suggesting that to retain the activity. According to these results, although some branching was present in the oligo-mannan, but the linear 1-3 linked glycosidic linkage was predominate in the active oligo-mannan. Conclusions: To retain the activity, the DP of the oligo-mannan should be greater than 6.
| P-06: Cloning and Functional Analysis of Key Genes TSA Involved in Indole Alkaloid Biosynthesis From Baphicacanthus Cusia | | |
Zhi-Ying Guo a,b, He-Xin Tan b, Xun-Xun Wu a,b, Yu-Xiang Huanga,b, Qing-Lei Zhang a,b, Yong Diao a and Lei Zhang b
a Institute of Molecular Medicine, Huaqiao University, Chenghuabei Road 269, Fengze District Quanzhou 362000, China
b Department of Medicinal Botany, School of Pharmacy, the Second Military Medical University, GuoHe Road 325, Shanghai 200433, P. R. China
Correspondence: Yong Diao, e-mail: [email protected]; Lei Zhang, e-mail: [email protected]
Introduction: Function analysis of the key gene TSA involved in indole alkaloid biosynthesis from Baphicacanthus cusia. Methods: The key gene involved in indole alkaloid pathway was systematically isolated based on the transcriptome sequencing data of Baphicacanthus cusia. The open reading frame of BcTSA gene was obtained by RT-PCR; through bioinformatics methods, the secondary structure and three-dimensional structure of these proteins were predicted and analyzed. The expression profile of the BcTSA gene in specific tissues and under induction with MeJA, SA and ABA were also observed by real-time quantitative PCR. Complementation of E.coli trpA mutant with the BcTSA cDNA to further study the biological function of TSA. Results: With RT-PCR, we cloned a novel gene BcTSA contains 945 bp open reading frames encoding a protein of 315 amino acids which belongs to the Trp_syntA superfamily. Sequence homology analysis of amino acid deduced from the cDNA showed that the conserved domains of BcTSA shared high homology with sixteen homologous genes. The spatial analysis results by quantitative RT-PCR showed that BcTSA was mainly expressed in stem and the chemical inducers of SA, MeJA and ABA can up-regulate the expression level of BcTSA. E.coli trpA mutant transformed with the BcTSA cDNA had ability to grow without tryptophan on minimal medium containing 0.02% arabinose supports the conclusion that this protein can catalyzes the formation of indole in Baphicacanthus cusia. Conclusions: The results demonstrated that BcTSA protein can catalyzes the formation of indole in Baphicacanthus cusia and might play an important role in indole alkaloid biosynthesis. | Figure 1: Alpha-tryptophan synthase (TSA) complemented the tryptophan synthase α-deficient
(9129, E.coli Δ trpA mutant ;TOPO, transformant containing the pBAD-TOPO vector; TSA, transformant containing the TSA open reading frame (ORF) in the pBAD-TOPO vector.)
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| P-07: Acupuncture and Related Therapies Versus Prokinetics for Functional Dyspepsia: Overview of Systematic Reviews and Network Meta-Analysis | | |
Robin ST Ho a, Xinyin Wu ab, Charlene HL Wong a, Justin CY Wu b, Vincent CH Chung ab
a Jockey Club School of Public Health and Primary Care, The Chinese University of Hong Kong, Hong Kong
b Hong Kong Institute of Integrative Medicine, The Chinese University of Hong Kong, Hong Kong
Correspondence: Robin ST Ho, e-mail: [email protected]
Introduction: Pharmacological interventions for functional dyspepsia (FD) have limited effectiveness. Acupuncture and related therapies could be used as an alternative. An overview of systematic reviews (SRs) and network meta-analyses (NMA) were performed to evaluate the comparative effectiveness of different acupuncture protocols, as compared to the prokinetics of domperidone, itopride, mosapride, and clebopride. Methods: We conducted a comprehensive literature search on SRs of randomized controlled trials (RCTs) in eight international and Chinese databases from their inception till November 2015. Methodological quality of included SRs was assessed. Data from RCTs were extracted for random effect pairwise meta-analyses. NMA was used to explore the most effective form of acupuncture when compared to domperidone, itopride, mosapride, and clebopride. Results: From five SRs, 22 RCTs (n=1727) assessing various different acupuncture protocol were included. Methodological quality of included SRs was mediocre. No serious adverse events were reported in all included studies. Five clinically homogeneous pairwise meta-analyses showed that manual acupuncture have stronger effect in alleviating global FD symptoms compared to domperidone (6 RCTs) and itopride (3 RCTs). There are no significant differences between moxibustion and domperidone (3 RCTs), and between manual acupuncture plus moxibustion and domperidone, as well as electroacupuncture and itopride (2 RCTs). Results from NMA showed that manual acupuncture plus moxibustion added on clebopride has the highest probability (95.0%) of being the best option for alleviating patient reported global FD symptom. This is followed by manual acupuncture plus moxibustion added on domperidone (76.1%), clebopride (74.5%), manual acupuncture (62.6%), moxibustion (62.3%), manual acupuncture plus moxibustion added on mosapride (61.9%), electroacupuncture (48.6%), manual acupuncture plus moxibustion (35.9%), domperidone (18.4%), mosapride (7.6%), and itopride (7.1 %). Conclusions: Manual acupuncture and moxibustion added on clebopride might be the most effective protocol for managing FD symptoms. FD patients should be avoid taking prokinetics, who are contraindicated with their potential side effects. Manual acupuncture could be an alternative to prokinetics for managing FD. Future confirmatory comparative effectiveness trials should use appropriate sham control and validated outcome measures beyond patient reported effectiveness.
| P-08: Antioxidant and Immune Capacity of Polysaccharide SMWP-U&E Isolated From Salvia Miltiorrhiza Bunge Residue In Vitro and In Vivo | | |
Yuan-Yuan Jiang a, Chao Mou b, Li Zhang a, Ying-Lun Li b, Yun-Song Zhang a
a College of Science, Sichuan Agricultural University, Yaan 625014, China
b College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang 611130, China
Correspondence: Li Zhang, e-mail: [email protected]
Objective: To investigate the antioxidant and immune capacity of SMWP-U&E, a kind of polysaccharides isolated from Salvia miltiorrhiza Bunge residue. Methods: The antioxidant ability of SMWP-U&E was evaluated in vitro by reducing power and scavenging free radical activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, superoxide anion radical and hydroxyl radical assays. Splenocyte proliferation was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to investigate the immune capacity of SMWP-U&E in vitro. To confirm the immune ability in vivo, carbon clearance assay and hemolytic assay were performed in mice. Further, the contents of cytokines including interleukin (IL)-2, IL-12, interferon (IFN)-γ and tumor necrosis factor (TNF)-α, and T lymphocyte subsets in serum of immunodepressive mice induced by cyclophosphamide were detected. Results: The results has shown that SMWP-U&E exerted strong reducing power and scavenging free radical activities in vitro on DPPH free radical (IC50=0.04 mg/mL), superoxide anion radical (IC50=0.24 mg/mL) and hydroxyl radical (IC50=0.53 mg/mL). It improved the proliferation of B-lymphocytes and T-lymphocytes cooperated with lipopolysaccharide (LSP) and concanavalin (Con A), respectively. Increase in phagocytic activity and half hemolytic value (HC50) was also observed in the present study. Meanwhile, the treatment with SMWP-U&E increased the contents of IL-2, IL-12, IFN-γ and TNF-α, as well as the CD4+ cell count and CD4+/CD8+ ratio in serum of immunodepressive mice induced by cyclophosphamide. Conclusions: SMWP-U&E has shown significant antioxidant capacity and also can improve the immunity.
| P-09: Salvia Miltiorrhiza BUNGE – 1H-NMR Metabolomics Approach in TCM Quality Control | | |
Ka Yui Kum a, Anthony Booker a, Christine Leon b, José Prieto García a, Michael Heinrich a
a Biodiversity and Medicines Research Cluster, UCL School of Pharmacy, London WC1N 1AX, U.K.
b Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, U.K.
Correspondence: Ka Yui Kum, email: [email protected]
Introduction: Many Danshen (DS - Radix Salviae miltiorrhizae - SM) products are found on the global market albeit without suitable quality assurance. Some substitutes including Salvia przewalskii Maxim, S. bowleyana Dunn. and Salvia sinica Migo. have similar chemical composition to SM. [1] This project aims at understanding the chemical variability among DS products associated with different value chains sold on different TCM markets. Methods: Ninety-seven DS products were sampled from different types of retailers and analysed by 1H-NMR spectrometry (followed by PCA analysis) in combination with HPTLC. Results: Out of twenty-eight DS final products, seven most likely-adulterated samples were found and of six these samples were collected from online stores. Another six samples of final products did not contain either salvianolic acid B or tanshinone IIA which are the DS chemical standards in most pharmacopeias. 1H-NMR results showed the chemical composition of Salvia bowleyana authenticated sample was similar to SM. Salvia przewalskii did not contain traceable amount of salvianolic acid B. Conclusions: The inconsistency of DS’s quality should be seen as an alarming signal relevant for TCM quality control and specifically how these products are processed, collected and enter the value chains.
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| P-10: Anti-Angiogenic and Immunomodulatory Activities of a Labdane Diterpene from Leonurus Heterophyllus | | |
Grace Gar-Lee Yue a,b, Julia Kin-Ming Lee a,b, Hin-Fai Kwok a,b, Da-Rong Wu c, Shao-Hua Liu c, Wei-Lie Xiao d, Clara Bik-San Lau a,b
a Institute of Chinese Medicine
b State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
c Chengdu No. 1 Pharmaceutical Group Co., Ltd., Chengdu 610031, China
d State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China
Correspondence: Clara Bik-San Lau, e-mail: [email protected]; Wei-Lie Xiao, e-mail: [email protected]
Introduction: Heteronone B (HB) is a labdane diterpene isolated from Leonurus heterophyllus Sweet which is known as Yimucao in Chinese medicines and is traditionally used for the treatment of gynaecological problems. HB was firstly isolated a decade ago but its biological activity has never been reported. Hence, the objectives of this study were to investigate the effects of HB on angiogenesis and immunomodulation using zebrafish embryos and human peripheral blood mononuclear cells (PBMCs), respectively. Methods: Angiogenesis in transgenic zebrafish Tg(fli1:EGFP)y1 was assessed by measuring the length of sub-intestinal vessels of embryos treated with HB. The mRNA expressions of angiogenic receptors in HB-treated zebrafish embryos were also examined. Besides, the proliferation and cytokines productions of PBMCs treated with HB were determined by thymidine incorporation assay and ELISA, respectively. Results: HB (5-10 μ M) was shown to significantly inhibit the growth of sub-intestinal vessels of zebrafish embryos without observed toxicity. In addition, it could down-regulate the mRNA expressions of angiogenesis-related genes in zebrafish embryos. Results on human PBMCs showed that HB (6.25-50 μ M) suppressed the proliferation of phytohaemagglutinin (PHA)-activated PBMCs. The levels of IL-10, IL-12, TNF-α and IFN-γ were modulated in PHA-activated PBMCs after HB treatment for 24 hours. Conclusions: This is the first report of the biological activities of HB showing the anti-angiogenic activities in zebrafish embryos as well as immunomodulatory activities in PBMCs. Our findings indicate that heteronone B may also act as an active component in this herb.
| P-11: An In Vitro Platform for Screening Chinese Herbal Medicines that Inhibit Cancer Stem-Like Cells in Breast Cancer | | |
Hoi-Wing Leung a,b, Chun-Hay Ko a,b, Grace Gar-Lee Yue a,b, Clara Bik-San Lau a,b
a Institute of Chinese Medicine
b State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
Correspondence: Clara Bik-San Lau, e-mail: [email protected]
Introduction: With the high recurrence rate of breast cancer, cancer stem-like cells (CSCs) are believed to be one of the possible cause of cancer recurrence. CSCs are subpopulation among cancer cells that possess stem cell characteristics: self-renewal and differentiation, and responsible for tumour’s cellular heterogeneity. Conventional therapies usually target on growing cancer cells rather than CSCs, so cancer recurrence may occur. The present study aimed to establish an in vitro platform for screening Chinese herbal medicines or natural products that inhibit the activities of CSCs. Methods: Breast CSCs-enriched cancer cells were developed from parental human breast cancer cell line MDA-MB-231 using two cell culture conditions: (1) culture medium supplemented with various growth factors particularly for CSCs (specific stem cell medium); (2) culture medium and low dose of doxorubicin. Results: The use of specific stem cell medium is preferred than that of culture medium and low dose of doxorubicin, since CSCs-enriched MDA-MB-231 cells exhibited higher self-renewal abilities and tumorigenicity, as compared with parental MDA-MB-231 cells. Our results also demonstrated that Chinese herb Hedyotis diffusa and its isolated compound inhibited spheroid formation of CSCs-enriched MDA-MB-231 cells. Conclusions: This screening platform can be applied to identify Chinese herbal medicines as potential anti-cancer agents that inhibit the activities of CSCs.
| P-12: Hypopigmentation and Healthy Skin Promoting Effect of a Modified Qi Bai San | | |
Elaine Wat a, Yau Tsz Chan a, Tiffany Hoi Ka Tso a, Chun Fai Ng a, Ching Po Lau a, Wing Sum Siu a, Chun Hay Ko a, Ka Keung Lau b, Raymond Hu b, Clara Bik-San Lau a, Ping Chung Leung a
a Institute of Chinese Medicine and State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
b 5100 Cosmetic Company Ltd., 130A Kwan Tei North Village, Fanling, New Territories, Hong Kong
Correspondence: Clara Bik-San Lau, e-mail: [email protected]
Introduction: Qi Bai San (QBS, 七白散) is a traditional Chinese herbal formula used by ancient ladies for healthy skin and whitening. Although it is a popular product in the market, it contains animal drug and toxic herbs which are undesirable. There is also lack of scientific evidence towards its efficacy. Hence, the aim of this study was to investigate whether a modified form of such classical herbal formula (containing dried rhizome of Atractylodes macrocephala 白术 dried sclerotium of Poria cocos 白茯苓, etc.) would exert beneficial effects on skin condition. Methods: MTT assay was performed to determine the non-cytotoxic concentration of this modified QBS water extract on murine B16 melanoma cells. Cellular melanogenesis and tyrosinase activity assays were used to assess the melanin content and tyrosinase activity, respectively, on a-melanocyte-stimulating hormone (MSH)-induced B16 cells. Collagenase inhibition assay was used to test if the extract could exert collagenase inhibitory activity. Results: The modified QBS extract (0.25–1 mg/ml) could significantly reduce α-MSH-induced increase in melanin content. This extract (0.5–1 mg/ml) could also dose-dependently reduce a-MSH-induced increase on tyrosinase activity, though not statistically significant. Besides, 1 mg/ml QBS extract significantly inhibited collagenase activity. Conclusion: We showed for the first time that a modified QBS formula could exert dose-dependent hypopigmentation and collagenase inhibitory effects. These in vitro data provided scientific evidence supporting the efficacy of this formula on hypopigmentation and healthy skin promotion.
Acknowledgement: This work was financially supported by Innovation and Technology Commission HKSAR, Innovation and Technology Support Programme (Collaborative project) Ref. no. ITS/164/14FX.
| P-13: Computational Identification and Systematic Analysis of the Dirigent Multigene Family in Salviae Miltiorrhizae | | |
Qing Li a, Jin-Yang Ma b, Jin-Dong Yang b, Lei Zhang b
a Department of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, No. 415, Fengyan Road, Huangpu District, Shanghai, 200003, China
b Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, No. 325, Guohe Road, Yangpu District, Shanghai, 200433, China
Correspondence: Qing Li, e-mail: [email protected]
Objective: To discovery and analysis the dirigent (DIR) and DIR-like protein gene family in Salvia miltiorrhiza (SmDIR). Methods: 33 DIR-like genes were found from the transcription profiling database rebuilt with 9 released transcription profiling of S. miltiorrhiza in GenBank. Among the 33 DIR-like genes, five are full-length sequences while the others are partial. The composition of nucleotide sequences and amino acid sequences, leader peptide, signal peptide, transmembrane topological structure, hydrohobicity or hydrophilicity, secondary structure, tertiary structure [Figure 1] and functional domain of the five full-length proteins were analyzed by bioinformatics methods. Homology analysis and phylogenetic trees of SmDIRs were also carried out. Results: The results showed that the open reading frames of the five SmDIRs were about 594 bp and the molecular weight were approximately 21.47 kDa. Except SmDIR5, the isoelectric point of the rest SmDIRs was lower than 7, between 5.60 and 6.50, which illuminated that most SmDIRs were acidic. Thr, Val, Gly, Leu and Ser were the major amino acids in all these proteins. The results of the sequence structure prediction show that SmDIRs have apparent hydrophobic region and hydrophilicity region, signal peptide and transmembrane topological structure. The main secondary structures of the proteins were random coil. Homology analysis reveals that the homology of SmDIR4 was high with Erythranthe guttata, while SmDIR1, SmDIR2, SmDIR3 and SmDIR5 were high with Sesamum indicum. Compared with Arabidopsis, SmDIRs were classified into 2 groups in the phylogenetic tree, DIR-b and DIR-d. Conclusions: Dirigent proteins are members of a large family of proteins which take important parts in the biosynthesis of lignin. However, the SmDIR protein genes have not been analyzed in detail yet. This work provides a systemic sequence analysis of SmDIRs. It will provide useful information for SmDIR studies in the future.
| P-14: Analysis on Inherent Relationships Between Herbs in Prescriptions of Prevention and Cure for Flu by Using TCM Inheritance System | | |
Zhi-Geng Li a, Yan Feng b, Shi-Huan Tang c
a Institute of Basic Chinese Medicine, China Academy of Chinese Medical sciences, Beijing 100700, China
b Beijing Electric Power Hospital, Beijing 100073, China
c Institute of Chinese Materia Medica, Academy of Chinese Medical Sciences, Beijing 100700, China
Correspondence: Zhi-Geng Li, e-mail: [email protected]
Objective: Using TCM inheritance system to analyze the inherent relationships between herbs in prescriptions of prevention and cure for flu. Method: Collect the prescriptions used for prevention and cure for flu in CNKI, then by using revised mutual information and complex system entropy cluster in the TCM inheritance system to analyze inherent relationships between herbs in prescriptions. Results: Based on the analysis of 132 cases of prescriptions in database, the frequency of each herb included in the database is computed, core combinations of herbs and relationship framework between herbs are mined from the database. Conclusions: TCM inheritance system can be used to analyze the prescriptions database and transfer complicated inherent relationships between herbs in prescriptions orderly. So it can provide powerful reference to researcher for decision. It has good applied value for the research of traditional Chinese medicine.
| P-15: The AP2/ERF Transcription Factors Regulate Active Ingredient Biosynthesis in Isatis Indigotica Fort. | | |
Rui-Fang Ma a,b,c, Ying Xiao a, Lei Zhang b, Yun Wang a, Jun Yinc and Wan-Sheng Chen a
a Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
b Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
c Development and Utilization Key Laboratory of Northeast Plant Materials, School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China
Correspondence: Jun Yin, e-mail: yinjun200[email protected]; Wan-Sheng Chen, e-mail: [email protected]
Objective: To investigate the characterizations of AP2/ERF transcription factors Ii49 and Ii99 and whether they could regulate the accumulation of lignans in I. Indigotica. Methods: Promoters of Ii49 and Ii99 was obtained by Genome walking. Expression levels of Ii049 and Ii099 in various tissues of I. indigotica with stress treatment were examined by quantitative real-time PCR (qRT-PCR). Protoplast transient transformation was used to observe the location of Ii049 and Ii099. The binding capability between Ii099 and GCC-box were assessed by electrophoretic mobility shift assay (EMSA). Ii049 and Ii099 silenced transgenic hairy roots were used to explore whether the biosynthesis of lignans and salicylic acid were regulated by Ii049 and Ii099. Overexpressing either transcription factor in order to obtain high quality herb with abundant lignans production. Relative expression of lignans biosynthetic genes in transgenic lines were detected by qRT-PCR. HPLC/MS/MS was used to determinate the accumulation of lignans after induced by exogenous salicylic acid. Results: Both promoters of Ii049 and Ii099 have JA and SA responsive elements. Ii049 and Ii099 were expressed in roots, stems and leaves with the highest expression in roots. Following stress treatments, Ii049 and Ii099 were significantly induced by Methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA). They were located in the nucleus when transiently expressed in rice protoplasts. Ii099 could binding to GCC-box specificly. Ii049 silenced transgenic hairy significantly inhibited lignans and SA accumulation while Ii099 silenced transgenic hairy enhanced lignans and SA contents significantly when compared with the wild type. By contrast, the contents of lignans and SA were increased in Ii049 overexpression lines while reduced in Ii099 overexpression lines. The expression of key enzyme also dramatically change in transgenic lines. SA could induced the accumulation of lariciresinol, but the effect wasn’t lasting. Conclusions: These results indicate that both Ii049 and Ii099 are MeJA and SA mediated key component of the molecular network regulating the accumulation of bioactive components in I. Indigotica, and Ii049 is one positive regulators while Ii099 is one negative regulators of lignans biosynthesis. | Figure 1: Lignans contents in transgenic hairy roots (*P < 0.05 vs. WT, **P < 0.01 vs. WT). A: Ii049 RNAi transgenic hairy roots, B: Ii099 RNAi transgenic hairy roots, C: Ii049 overexpress transgenic lines, D: Ii099 overexpress transgenic lines
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| P-16: Harmful Plant Combination: Implication of the Traditional Chinese Herb Hou Po (Magnolia Officinalis) in the Cytotoxicity and Genotoxicity of Aristolochia Sp. | | |
Melanie Poivre a, Amandine Nachtergael a, Marie-Hélène Antoine b, Joëlle Nortier b Anastasia Atsapkina c, Kirill Kryshenc, Alexander Shikov c, Pierre Duez a
a Unit of Therapeutic Chemistry and Pharmacognosy, Faculty of Medicine and Pharmacy, Research Institute for Health Sciences and Technology, University of Mons (UMONS), 20 Place du Parc, 7000 Mons (Belgium)
b Laboratory of Experimental Nephrology, Faculty of Medicine, Université Libre de Bruxelles, Brussels, Belgium
c Saint-Petersburg Institute of Pharmacy, Leningrad region, Vsevolozhsky district, 188663, Kuzmolovo P 245, Russia
Correspondence: Melanie Poivre, e-mail: Mé[email protected]
Introduction: Traditional Chinese herbs, known for thousands of years, are often considered by the general public as “safe and reliable”. However, herbs can sometimes cause serious health troubles. In some cases, plants combinations can also be very harmful. For example, the “Chinese herbs nephropathy”, a progressive renal interstitial fibrosis, was reported in a Belgian cohort after the intake of slimming pills containing the bark of Magnolia and also inadvertently the roots of an Aristolochia species. Material and methods: This work evaluated in vitro (intestinal and renal human cells and bacterial models) the cytotoxicity and genotoxicity of the two plants Aristolochia baetica and Magnolia officinalis and their association. Cytotoxicity was studied by MTT and resazurine cell viability assays and the genotoxicity by the detection and quantification of the phosphorylated histone γ-H2AX (whole cell ELISA and Immunocytochemistry) and by an Ames test on four different Salmonella typhimurium strains (TA98, TA100, TA1535 and TA1537). Using analytical methods (HPTLC and MS) for plants characterization, the present work also tends to determine which Magnolia compounds are responsible for the high potentiation of AAs genotoxicity and cytotoxicity. Results: The genotoxicity of the association Aristolochia baetica - Magnolia officinalis is significantly higher (at least 3 biological replicates, p<0.001), compared to the individual plant extracts in both γ-H2AX detection and Ames test. Discussion: Although a very interesting example of indirect genotoxicity, this observation warrants further studies to establish the precise role of the Magnolia officinalis extracts and to identify compounds and mechanisms responsible for the observed co-genotoxicity.
| P-17: Chemical Constituents from Hericium Erinaceus | | |
Xiao-Juan Zhang a, Yang Chen a, Wei Wang a and Gulnar Sabir b
a TCM and Ethnomedicine Innovation & Development Laboratory, School of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan,410208, China
b Xinjiang Institute of Chinese Material Medica and Ethica, Urumqi Xinjiang ,830002, China
Correspondence: Gulnar Sabir , e-mail: [email protected]
Since a cultivation technique applied on the fungus bed, using bags and bottles, fungus ranked among the cultivating mushroom available on the market. Hercium erinaceum is one of the edible mushrooms widely distributed in China and Japan [1]. This Basidiomycetesis is found growing on rotting hardwood trees such as Pterocarpus indicus, Cyclobalanopsis spp. and Fagus crenata. When air-dried, it is used as an herbal medicine called the “Houtou”, extracted with hot water or brewed in hot water, which considered as a health drink. Hercium erinaceum exerted anti-inflammatory, anti-skin aging, anti-cancer, anti-diabetic, anti-hypertensive, anti-microbial, cardio-protective, hepato-protective, immunomodulatory, and neuro-protective effects. It have been reported to produce several classes of bioactive molecules like hericenones and erinacines. The purpose of our project is to isolate the low molecular constituents towards to the anti-tumor and immunomodulating activities from the ethyl acetate extract of Hercium erinaceum.
Acknowledgements: Support for this research from the Graduate student innovation project in Hunan province (CX2016B358) is gratefully acknowledged.
| P-18: A New HPLC Method for the Quantitative Analysis of Harpagide and Harpagoside in Scrophulariae Radix: Monograph Proposal for the German Pharmacopoeia | | |
Katharina Schiller a, Jorg Heilmann a, Detlef Manns b, Gerhard Franz a
a Pharmceutical Biology Department, University of Regensburg, Universitätsstraße 31, 93053 Regensburg, Germany
b Federal Institute for Drugs and Medical Devices (BfArM), Kurt-Georg-Kiesinger-Allee 3, 53175 Bonn, Germany
Correspondence: Katharina Schiller, e-mail: [email protected]
Introduction: The increasing interest in Traditional Chinese Medicine and the lack of quality monographs in the German Pharmacopoeia is the basis for research about the characterization of the most important herbal drugs, which are commonly used in TCM, to guarantee safety and quality. Scrophulariae radix (Xuanshen) is the dried root of Scrophularia ningpoenis HEMSL., with iridoids to be known as the major chemical constituents [1]. Hence, harpagide and harpagoside were selected to be quantified. Material and Methods: Quantification was performed by HPLC on a RP-18e column and detected by DAD (210 nm). The mobile phase was composed of aqueous phosphoric acid (0.03%, V/V) and acetonitrile using a gradient elution (flow rate 1 mL/min). Results: Accuracy (recovery), precision (RSD ≤ 5%), specifity (resolution ≥ 1) and linearity were proven. Furthermore, robustness of the method was investigated. Conclusion: A new HPLC method based on the Pharmacopoeia of the People’s Republic of China was developed and validated in order to quantifiy the two iridoids according to the ICH-Guidelines [2].
Acknowledgements: The BfArM is gratefully acknowledged for financial support.
| P-19: Combined Contaminant and Multi-Residue Testing by LC/MS/MS for the Quality Control of Herbal Drugs Used in Traditional Chinese Medicine (TCM) | | |
Thomas Glauner a, Morgane Even a, Tao Bo c, Andreas Tei a, Lester Taylor b
a Agilent Technologies R&D and Marketing GmbH, Hewlett-Packard Str. 8, 76337 Waldbronn, Germany
b Agilent Technologies, Inc.,5301 Stevens Creek Blvd, Santa Clara, CA 95051, USA
c Agilent Technologies, No.3, Wang Jing Bei Lu, Chao Yang District, Beijing 100102, P. R. China
Correspondence: Andreas TEI, e-mail: [email protected]
Introduction: Routine screening and quantitation for pesticides is important to increase the safety of herbal drugs used in TCM. Medicinal plants are also affected by fungal infections and the presence of weed plants raising the importance of testing for natural toxins. In this presentation we show a new rapid routine approach for combined screening and quantification of pesticides, mycotoxins and pyrrolizidine alkaloids in herbal drugs. Materials and Methods: A total of 10 different dried herbal drugs were homogenized and extracted using QuEChERS without clean-up. The extract was chromatographically separated on an Agilent 1290 Infinity UHPLC system combined with an iFunnel triple quadrupole mass spectrometer operated with dynamic MRM and fast polarity switching. The final method included 267 pesticides, 11 mycotoxins and 28 pyrrolizidine alkaloids. For method performance characterization samples were spiked with all target compounds at relevant concentrations. Results: Low levels of detection have been achieved by the optimized UHPLC method in combination with the highly sensitive triple quadrupole LC/MS system. Limits of quantitation for most pesticides were below 10% of their maximum residue limit even with higher dilutions. With a 10 to 20-fold sample dilution, matrix effects were minimized and accuracies for most of the targeted compounds were within the desired range of 70 to 120%. Several pesticides and pyrrolizidine alkaloids were identified in the herbal drugs in concentrations of 10 to 100 μg/kg. All results will be presented. Conclusions: Fast routine screening using state of the art LC/MS/MS instrumentation is an efficient approach to increase the safety of herbal drug material used in TCM and to meet regulatory safety standards.
| P-20: Peculiarity of use of Species of Local Flora Finno-Ugric Ethnic Group as Medicinal Plants | | |
Lebedeva T.P. a, Tkachenko K.G. b
a Herbarium of the Komarov Botanical Institute of RAS, St. Petersburg 197376, Russia
b Peter the Great Botanical Garden of the Komarov Botanical Institute of RAS, St. Petersburg 197376, Russia
Correspondence: Lebedeva Tatjana, e-mail: [email protected]; Tkachenko Kirill, e-mail: [email protected]
Objective: The aim of this work was to collect and study materials ethnobotanical number of Baltic and Finnish ethnic peoples of the North European part of Russia (Veps, Karelians, Izhora, Vodes, Estonian-Seto), Komi-Zyrian, Saami and Russian, with which almost all of these people have long live in more or less close geographical and cultural contact. Methods: There was a list of 160 questions on the use of local flora plants as useful. Results: It was found that the most common herbs like almost all of the small peoples of the North used birch – Betula sp. (juice, buds, leaves) cloudberries – Rubus chamaemorus L. (sepals), cranberries – Vaccinium vitis-idaea L. (leaves), cranberry – Vaccinium oxycoccos L. is a synonym of Oxycoccus palustris Pers. (fruit). Very rarely, some population groups use plantain – Plantago major L. (leaves), yarrow – Achillea millefolium L. (herb) and nettle – Urtica dioica L. (leaf). The fruit of dog-rose (or canker-rose) – Rosa sp. using very rare only for herbal-tea, not as a medicinal raw material. No one uses as a medicinal raw rowan – Sorbus aucuparia L. (fruit), may-lily – Convallaria majalis L. (leaves) although this type of raw material included in the Pharmacopoeia and uses in official medicine. Origanum vulgare L., Hypericum perforatum L., Mentha longifolia (L.) L., and/or M. arvensis L. – using very rare, and only as one of component of herbal tea. The given data clearly show that different people, even close to living one kind of plants (and parts thereof) are not always the same. Even one of the people, but living in a certain geographic location, so do not always use the same one vegetable raw material. Russian penetration into the habitat of indigenous peoples does not always lead to the fact that they are all used for the same purposes the same plant material. Conclusions: The survey informants, representatives of various small peoples of the North has revealed that they are not using many species of local flora as medicinal plants, although some of the bottom on the list of permitted officially for use in medicine.
| P-21: Screening the Effective Parts of Sanggua Decoction Hypoglycemic Effects in Type 2 Diabetic Rats | | |
Qi-Chang Xing, Ying Wang, Fei Zhi, Yu Cai, Yun-Zhong Chen
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China
Correspondence: Yun-Zhong Chen, e-mail: [email protected]
Objective: To select the effective parts of hypoglycemic effects in type 2 diabetic rats on Chinese herbal compound Sanggua Decoction (SGD). Method: 1.Male SD rats were constructed by high-fat-diet and combined with intraperitoneal injection of streptozocin 40 mg kg-1,whose fasting blood glucose (FBG) greater than 11.1 mmol/L is successful DM mold and the model rats were randomly divided into 9 groups; 2.Use the ethyl acetate and n-butanol to extract SGD concentrated solution in order, then get the ethyl acetate fraction (E), n-butanol fraction (N) and the aqueous parts of the group (A), the writer set up the E+N,E+A, N+A,E+N+A groups (raw herb 8.3 g ·kg-1) in addition, positive control group (PC) (metformin 0.15 ·kg-1), blank group(B) (normal saline) was also designed; 3.The changes of the FBG was observed in time, after 6-week treatment, take the eyeball for blood and separate the serum, measure the glycosylated hemoglobin (HbA1c) then kill the rats and dissect, cut a part of the liver and weigh it, test the amount of hepatic glycol, observe the histological sections of pancreas. Result: Compared with group B, groupN+A could decrease FBG by 66.9%, decrease HbA1c by 13.6%, increase the amount of hepatic glycol by 22.5%, the histological sections of pancreas is the best. Conclusion: The mixture of n-butanol fraction and aqueous parts of the group is the best effective part, coordinatingly synergic effect during their hypoglycemic need to be further studied.
| P-22: LC/MS/MS Analysis of the Metabolites Of Lathyrane Diterpenoids in CACO-2 Cells | | |
Si-li Tang a, Min-ting Lin a, Ling-ling Zhang a, Wen-jing Huang a, Yun Liu a, Zhong-zhen Zhao b, Hu-biao Chen b, Jian-ye Zhanga
a Guangzhou Medical University, Guangzhou 511436, China
b School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China
Correspondence: Jian-Ye Zhang, e-mail: [email protected]
Objective: To identify the metabolites of Euphoria Factor L1, L2, L8 and 6(17),12(E)-lathyrol-5,15-diacetate-3-phenylacetate in Caco-2 cells by LC/MS/MS. Methods: Caco-2 cells were cultured with 100 μM lathyrane diterpenoid for 3, 6, 12h, respectively. Then the samples were collected, purified and identified by LC/MS/MS. Results: The major metabolites of Euphoria Factor L1 were two methylated products which were obtained after hydrolysis of the ester. The major metabolites of Euphoria Factor L2, L8 and 6(17),12(E)-lathyrol-5,15-diacetate-3-phenylacetate were hydrolysis products of the ester. Conclusions: The main metabolic pathway of Euphoria Factor L1 was methylation and hydrolysis of the ester. The main metabolic pathway of Euphoria Factor L2, L8 and 6(17),12(E)-lathyrol-5,15-diacetate-3-phenylacetate was hydrolysis of the ester. LC/MS/MS can identify the metabolites of Euphoria Factor L1, L2, L8 and 6(17),12(E)-lathyrol-5,15-diacetate-3-phenylacetate in Caco-2 cells quickly and sensitively.
| P-23: The Protective Effect of Rhizoma Dioscoreae Extract Against Alveolar Bone Loss in Ovariectomized Rats Via Regulating IL6/STAT3 Signaling | | |
Zhiguo Zhang a, Zhigeng Li a, Changheng Song a, Fangzhen Zhang a, Lihua Xiang a, Yanjing Chen a, Gary Guishan Xiao b and Dahong Ju a
a Institute of Basic Theory, China Academy of Chinese Medical Sciences, Beijing 100700, China
b School of Pharmaceutical Science, Dalian University of Technology, Dalian 116024, China
Correspondence: Dahong Ju, e-mail: [email protected]
Introduction: The aim of this study was to evaluate the osteoprotective effect of aqueous Rhizoma Dioscoreae extract (RDE) on the alveolar bone of rats with ovariectomy-induced bone loss. Materials and Methods: Female Wistar rats were subjected to either ovariectomy or a sham operation (SHAM). The ovariectomized (OVX) rats were treated with vehicle (OVX) or RDE by oral gavage or with 17β-estradiol (E2) subcutaneously. After treatments, the bone mineral density (BMD), the three-dimensional bone architecture of the alveolar bone and the plasma biomarkers of bone turnover were analyzed to assess bone metabolism, and the histomorphometry of the alveolar bone was observed. Rat IL6/STAT3 signaling pathway PCR array were used to evaluate differentiation of gene expression in alveolar bone from RDE-treated, OVX rats and SHAM rats. The differential expression of genes was verified using real-time quantitative RT-PCR (qRT-PCR). Results: Our results showed that RDE inhibited alveolar bone loss in OVX rats. Compared to the OVX rats, the RDE-treated rats showed downregulated expression levels of 20 genes and upgulated expression levels of 1 gene in the alveolar bone. Conclusions: These experiments revealed that RDE could inhibit ovariectomy-induced alveolar bone loss in rats. The mechanism of this anti-osteopenic effect in alveolar bone may be involved in the reduced abnormal bone remodeling, which is associated with the modulation of the IL6/STAT3 signaling pathway via gene regulation.
Acknowledgements: the National Natural Science Foundation of China (81473450) | Figure 1: Schematic diagram illustrating the IL6/STAT3 signaling in an anti-osteopenic effect of RDE. The downregulated genes appear in green. The white indicates the genes that are not specified, but that are incorporated into the network through relationships.
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| P-24: Profiling and Identification of Metabolites of Isorhynchophyllin in Rats By Ultra High Performance Liquid Chromatography And Linear Ion Trap Orbitrap Mass Spectrometry | | |
Jianwei Wang a,b,c, Peng Qi a, Jinjun Hou a, Yao Shen a, Bingpeng Yan a,b, Qirui Bi a,b, Ruihong Feng a, Xiaojian Shi a,b, Min Yang a, Wanying Wu a and De-an Guo a
a Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai 201203, China
b College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing 210009, China
c School of Chemical and Biological Engineering, Nantong Vocational University, Nantong, 226007, China
Correspondence: Wanying Wu, e-mail: [email protected]; De-an Guo e-mail: [email protected]
Introduction: The searching of potentially bioactive metabolites in the biological body is an interesting and meaningful work for drug study. However, the structural clarification of possible metabolites is one of the most challenging tasks in drug metabolism studies because of the variety of metabolic reactions and complexity of metabolites in vivo. To investigate profiling and identification of metabolites of isorhynchophyllin (IR) in vivo and vitro. Methods: An ultra high performance liquid chromatography/ linear ion trap-Orbitrap mass spectrometry (U-HPLC/LTQ-Orbitrap-MS) with combination of data post-processing techniques, including extracted ion chromatogram (EIC) and multiple mass defect filters (MMDF) was established, and the possible metabolic pathways were subsequently proposed after the oral dose of 20 mg/kg to male Wistar rats. The relative percentage of each metabolite and main metabolite types were also determined to better understand the metabolic behavior of IR in rats. Results: A total of 47 metabolites of IR were tentatively identified, including 47, 21, 18, and 25 metabolites in rat urine, plasma, liver and rat liver microsomes (RLM) samples, respectively. Conclusions: To our knowledge, most of them were reported for the first time. Seven metabolic pathways, including dehydrogenation, oxidation, hydrolysis, reduction, demethylation, hydroxylation and glucuronide conjugation were involved in the metabolism. Among them, dehydrogenation, hydrolysis, hydroxylation and oxidation were considered as the main metabolic pathway of metabolism in vivo and in vitro. The newly discovered IR metabolites significantly expanded our understanding and were going to be greatly helpful for the further pharmacokinetic study of IR.
| Chemical Characterization and Discrimination of Fruit, Stem and Leaf of Tribulus Terrestris L. by UPLC-QTOF-MS Based Profiling and Chemometrics Analysis | | |
Peng Qi a,1, Yuanyuan Zheng a,1, JinjunHou a, YaoShen a, Jianwei Wang a,b, Qirui Bia,b, RuiHong Feng a, Min Yang a, Wanying Wu a,*and De-an Guoa,b,*
a.Shanghai Research Center for Modernization of Traditional ChineseMedicine, National Engineering Laboratory for TCMStandardization Technology, Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Haike Road 501, Shanghai 201203,China b College of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing 210009, China
Correspondence: Wanying Wu, e-mail: [email protected]; Dean Guo, e-mail: [email protected]
1 These authors contributed equally to this work.
Objective: This study was carried out to overall profiling predominant constituents, steroidal saponins in fruits, stems, leaves of Tribulus terrestris L. (TT), and investigate the characteristic chemical markers for differentiating between them. Methods: Twenty batches of TT were collected from designated area and extracted properly. An integrated platform based on component profiling and chemometrics analysis was adopted. Firstly, the predominant constituents of TT, steroidal saponins in fruits, stems and leaves were rapid characterized by UPLC-QTOF/MS in negative ion modes. And the possible structures of new saponins were first deduced out by accurate masses and the characteristic fragmentation ions. After that, by using pattern recognition analysis, saponins identified above was extracted and aligned as dataset, and a clear separation of the three parts of TT was achieved as well as typical saponins were screened as the valuable variables for discrimination. Results: A total of 91 steroid saponins were characterized or tentatively identified distributed over eight steroidal aglycone skeletons, including 37 compounds reported for the first time from TT. With PCA and PLS-DA analysis, the three parts of TT were separated clearly and corresponding marker compounds were also identified. Additionally, the two saponin types, furostanolic and spirostanolic nucleus in TT were found distributed differently in three parts. That furostanolic nucleus are mainly in fruits and spirostanolic nucleus present in relatively high concentrations in stems and leaves. Conclusions: our results visualized the difference between fruits, stems and leaves of TT by effective components profiles and chemometrics, and this approach would be helpful for quality control of Chinese medicine, especially those with different medical parts.
| P-26: A Strategy Combining Higher Energy C-trap Dissociation With Neutral Loss- and Product Ion- Based MSN Acquisition for Global Profile and Structure Annotation Of Conjugated Bufadienolides in Venenum Bufonis | | |
Qi-rui Bi a,b,1, Jin-jun Hou a,1, Min Yang a, Yao Shen a, Peng Qi a, Rui-hong Feng a, Zhuo Dai a, Bing-peng Yan a,b, Jian-wei Wang a,b, Xiao-jian Shi a,b, Wan-ying Wu a and De-an Guo a,b
a Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai 201203, China
b The college of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing 210009, China
Correspondence: Wan-ying Wu, e-mail: [email protected] and De-an Guo, e-mail: [email protected]
1 These authors contributed equally to this work.
Objective: To globally profile and targeted structurally annotate of new minor toxic conjugated constituents, dicarboxylic acid conjugated bufadienolides (DACB), in Venenum Bufonis (VB). Methods: An integrated strategy (HPNA), combining higher-energy C-trap dissociation (HCD) and Msn (n≥3) acquisition based on product ion (PI) and neutral loss (NL) by LTQ-Orbitrap, was developed to study DACBs in extract of VB enriched by chromatographic method. The validation of the HPNA strategy were based on three new compounds which were purified by the LC/MS guided separation. Result: In all, 78 DACBs have been discovered from VB by the HNPA strategy, of which 68 were new compounds including 3 new isolated compounds, 52 tentatively identified compounds and 13 unknown compounds. 25 kinds of sub-structure formulas and seven kinds of dicarboxylic acid side chains were founded, especially for those five new side chains (two saturated dicarboxylic acids (azelaic acid (C9) and sebacic acid (C10)) and three un-saturated dicarboxylic acids (u-C8, u-C9, and u-C1o)), which have greatly enriched the structures of DACBs. Furthermore, the three new compounds were purified and identified by NMR. Conclusion: The result demonstrated that the HPNA strategy is a powerful tool for the global profile and structure annotation of known and unknown DACBs from VB.
| P-27: Simultaneous Quantification of Nine Alkaloids in Uncariae cum uncis Ramulus Using a Single Reference Standard by HPLC-UV Method | | |
Ruihong Feng, Jinjun Hou, Yibei Zhang, Huiqin Pan, Wanying Wu and De-an Guo
Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai 201203, China
Correspondence: Wan-Ying Wu, e-mail: [email protected]; De-an Guo, e-mail: [email protected]
Objective: To control the quality of Uncariae cum uncis ramulus., study the variations in the total content of alkaloids in different species and reveal the influence caused by rates and appearance of hooks. Methods: A HPLC-UV method was established for the quantitative analysis of nine indole alkaloids using a single reference standard to determine multi-components (SSDMC). Isorhynchophylline was selected as the reference compound for calculating the relative response factors and Uncaria rhynchophylla (Miq.) Miq. ex Havil was used to the establishment of methodology. The new assay method was fully validated in respect to precision, accuracy and repeatability. The proposed method was successfully applied to quantify the nine indole alkaloids in the other four recorded species mentioned in Chinese Pharmacopeia 2015. The same batch of crude drug was classified to four grades: pure stem, thin hooks, medium hooks and thick hooks. The caparison of them was conducted by the same method. Results: The results demonstrated significant variations in the total content and quantity of each of the nine indole alkaloids in five different species, indicating that reasonability of mixing up in clinical applications. The rates and appearance of the hooks had no influence on the total content of nine indole alkaloids. Conclusions: This simple, rapid, low-cost and reliable HPLC–UV method using SSDMC is suitable for routine quantitative analysis and quality control of Uncariae cum uncis ramulus.
| P-28: Secondary Metabolite Profiling-Based Multivariate Data Mining Approaches on the Discovery of Chemical Combination for the Characterization of Close-Related Herbal Medicines | | |
Wen Gao, Ke Liu, Hua Yang, Ping Li
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China
Correspondence: Ping Li, e-mail: [email protected]
Herbal medicines (HMs) are attracting intensive attention due to their long historical clinical use and reliable therapeutic efficacy at present. The authentication is the key for their safety and efficacy. The flowers of Lonicera japonica Thunb (“Jinyinhua” in Chinese, JYH) were extensively used in Eastern Asia, and some related species, such as L. macranthoides, L. fulvotomentosa, L. confusa, and L. hypoglauca (“Shanyinhua” in Chinese, SYH) have been confused or misused. In this work, Lonicera species samples were used as a typical example, contributes to the development of a novel strategy to discover the markers which appropriate for quality control. A matrix solid phase dispersion (MSPD) method was designed for the extraction of saponins, the major chemical difference between JYH and SYH. An ultra-high performance liquid chromatography (UHPLC)–tandem quadrupole time-of-flight mass spectrometry (QTOF MS) was applied for qualitative analysis and triple quadrupole mass spectrometry (QQQ-MS) method was developed for detection of 12 components. Then multivariate statistical analysis, such as principal componentanalysis (PCA), back propagation-artificial neural network (BP-ANN) model, and partial least square discriminant analysis (PLS-DA) model, were used for classification and prediction, and the contribution of each compounds to species classification was calculated. Using this strategy, three compounds are in combination contributed to species characterization of varied Lonicera samples were found and applied to their specific tests. This strategy provides an applicable approach to selection of chemical markers for quality control of closely herbal medicines.
| P-29: Effects of Abbiflorin, Paeoniflorin on the Behavior and Hypothalamic-Pitutary-Axis in Chronic Restraint Stress Depressed Rats | | |
Cheng-Long Wang a, Lin-Yuan Wang b, Jing-Xia Wang a, Ying-Li Zhu a, Dan-Ping Zhao b, Sha Wang a, Li Wu a, Jian-Jun Zhang a
a School of Basic Medical Science of Beijing University of Chinese Medicine, Beijing 100029, China
b School of Chinese Materia Medica of Beijing University of Chinese Medicine, Beijing 100102, China
Correspondence: Jian-Jun Zhang, e-mail: [email protected]
Objective: To observe the effects of Albiflorin (AF) and Paeoniflorin (PF) on behaviors and hypothalamic-pituitary-adrenal axis (HPAA) in chronic restraint stress (CRS) depressed rats and to explore the underlying mechanism. Methods: Male SD rats were randomly divided into 8 groups (n=12 in each). Except the normal control, the other groups were treated with the CRS stimulation combined with lonely. The behavior changes of rats were detected through open-field test, elevated plus-maze test and sucrose preference test. The levels of corticotrophin releasing hormone (CRH) in hypothalamus, adrenocorticotropic hormone (ACTH) and cortisol (CORT) in plasma were measured by radioimmunoassay. Results: On the 14th day, the weight in the 30 mg•kg-1 AF and 30 mg•kg-1 PF increased; on the 21st day, the sugar consumption, weight, cross count in the 30 mg•kg-1 AF and 30 mg•kg-1 PF increased, whereas stay period decreased; the time and stay period in the central zone, times and stay period of entering open arm increased, but the stay period in close arm decreased; the content of CRH, ACTH and CORT in the 30 mg•kg-1 AF and 30 mg•kg-1 PF were decreased. Conclusions: AF and PF are two main components of Paeoniae Radix Alba. They can improve, to some extent, the behaviors of CRS depressed rats. The mechanism might associate with to changing the stress hormone contents and inhibiting effects of HPAA activation. AF and PF are both the main effective components of Paeoniae Radix Alba.
Acknowledgements: The authors are grateful for financial support from National Nature Science Foundation of China (No.81473370) and Patent (No. ZL 201110184287.4, China).
| P-30: Neuroprotective Effects of Albiflorin Against Corticosterone-Induced Neurotoxicity in PC12 Cells Via No/Ros Signal Pathway | | |
Ying-li Zhu a, Lin-yuan Wang a, Jing-xia Wang a, Zhi-hui Yang b, Cheng-long Wang a, Zi-chen Wang a, Li Wu a, Jian-jun Zhanga
a Beijing University of Chinese Medicine, Beijing, 100029, China
b Department of Psychiatry & Neuroscience, University of Florida, Gainesville, FL 32611, USA
Correspondence: Jian-Jun Zhang, e-mail: [email protected]
Introduction: Albiflorin (AF), a monoterpene glycoside isolated from P. lactiflora, possess a variety of pharmacological activities, especially on psychiatric disorders. This study investigated whether AF(1mM, 0.5 mM, 0.25 mM) against corticosterone (CORT, 250 u M)-induced injury in PC12 cells and examined the potential mechanisms underlying. Methods: Cell viability and cytotoxicity were detected using a CCK8 assay. PC12 cell apoptosis was measured using Annexin V/PI and a DNA fragmentation assay kit, intracellular Ca2+ concentrations were assessed by fluorescent labelling, the intracellular reactive oxygen species (ROS) levels and nitric oxide (NO) were measured using 2’,7’-dichlorfluorescein-diacetate (DCFH-DA) staining and 3-amino,4-aminomethyl-2’,7’-difluorescein diacetate (DAF-FMDA). Next, the mitochondrial permeability transition pores (mPTPs) and mitochondrial membrane potentials (ΔΨm) were detected using a colorimetric mPTP detection kit and a 5,5’,6,6’-tetrachloro-1,1’,3,3’- tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) kit, respectively. Finally, neuronal nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), cytochrome c, caspase-3 expression levels were monitored by western blot analysis. Results: Treatment with CORT induced cytotoxicity in PC12 cells (P<0.001). AF dose-dependently increased cell viability (P<0.05, P<0.01). Compared with the CORT-treated group, AF slowed the accumulation of ROS (P<0.01) and NO (P<0.05, P<0.01), decreased the level of intracellular free Ca2+ (P<0.01), including mPTPs and ΔΨm. Furthermore, the down-regulation of cytochrome c (P<0.001, P<0.01) and the blockage of caspase-3(P<0.001, P<0.01) activity and inhibiting the overexpression of nNOS (P<0.01) and iNOS (P<0.01) were observed upon AF treatment. Conclusions: Our data show that the neuroprotective effect of AF may be attributed to the ROS/NO pathway.
Acknowledgements: The authors are grateful for financial support from National Nature Science Foundation of China (No.81473370) and Patent (No. ZL 201110184287.4, China).
| P-31: Nephrotoxic Botanicals: Phenotypic Framework and Mechanisms of Action | | |
Francesca Holden a and Qihe Xu b
a Medical student at King’s College London, Guy’s Campus, London SE1 1UL, United Kingdom
b Centre for Integrative Chinese Medicine and Department of Renal Sciences, King’s College London, The Rayne Institute, 123 Coldharbour Lane, London SE5 9NU, United Kingdom
Correspondence: Qihe Xu, e-mail: [email protected]
Introduction: Consuming aristolochic acid (AA)-containing plants is well established to cause renal damage and urothelial carcinoma. Unfortunately, these plants are not the only botanical nephrotoxicants. In this study, we aim to collate checklists and related evidence of plant materials linked to renal and urothelial injury. Methods: Through critical review of the literature, we propose a phenotypic framework for botanical-associated nephrotoxicity, illustrate mechanisms, highlight problems in the field and discuss future directions. Results: Botanical-associated nephrotoxicity can manifest as acute kidney injury, chronic kidney disease, glomerular injury, tubular dysfunction, nephrolithiasis and neoplasia. Though causal effects are well established for some, with offending compounds identified and supported by good clinical, in vitro and in vivo evidence, many others are associations that will need further investigation. Mechanisms of injury are obscure for many cases, but known mechanisms include direct cytotoxicity, fibrogenesis, carcinogenesis, blockading renal transport, inducing hyperoxaluria, causing hypertension, etc. Some botanical species cause nephrotoxicity without established safe dosage, thus should be banned or only use when benefits outweigh toxicity. Nephrotoxicity of many botanicals are due to over-dose, prolonged use, herb-herb or herb-drug interaction or wrong indications, therefore could be prevented by proper training and education of healthcare professionals and the public. Some other toxicities, however, are due to compromised quality of botanical products caused by contamination, adultery, etc, and could be prevented by more stringent quality control of botanical products. Conclusions: Botanicals are associated with a wide spectrum of nephrotoxicity through different mechanisms. Developing an ever updating, open-access online resource to catalogue the known phenotypes, mechanisms and the level of confidence of concerned associations will be of important clinical, scientific and regulatory values, and for the safety of the public. We hope that this work contributes to this ultimate goal.
Acknowledgements: This work is the outcome of a Year-3 MBBS student-selected component project supervised by Dr Qihe Xu at King’s College London. We thank Dr Jeffrey Kopp at NIDDK, NIH, USA, for his invaluable comments and thank Jack Lilly D’cruz, Niran Yoganayagam, Thurkka Raveendranathan and Vanisha Amin, as well as examiners of this library project for partially inspiring our work.
| P-32: Identification of Notoginsenosides of Fufang Xueshuangtong Capsule in Beagle Dogs by HPLC-LTQ-orbitrap-MSN Based on An Integrate Screening Approach | | |
Mei-fang Jiang, Huan-huan Pang, Qin-hui Wang, Jian-mei Huang
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100102, China.
Correspondence: Jian-mei Huang, e-mail: [email protected]
Objective: Fufang Xueshuangtong capsule (FXT) has been regarded as a preferred Chinese herbal formula for the treatment of Diabetic retinopathy (DR) and similar diseases. Notoginsenosides are the main active ingredients in the FXT. In this study, to remove the interferences of the diverse chemical composition of TCM and the complex matrix of biological samples, a novel and efficient strategy was developed and used for the screening and identification of notoginsenosides in vivo and in vitro using high performance liquid chromatography combined with LTQ-Orbitrap mass spectrometry (HPLC-LTQ-Orbitrap-MSn). Methods: The data mining techniques, the mass defect filtering (MDF) and the characteristics ion filter (CIF), were adopted for the rapid screening of notoginsenosides from complicated peaks, with the high resolution mass spectrometry. Furthermore, the screened notoginsenosides were identified by the accurate mass, MS/MS, and retention time (tR), obtained by the full scan and parent ion list (PIL) scan on HPLC-LTQ-Orbitrap, then compared with retrieving literatures and references. Results: By using this strategy, 46 notoginsenosides were totally screened and identified in vivo, and 18, 48, 15 notoginsenosides were respectively screened and identified in plasma, feces, urine, after oral administration of FXT in beagle dogs. Conclusions: This study laid the foundation for the material basis research of FXT and provided a new strategy for rapid identification of notoginsenosides.
| P-33: Comparative Study on Therapeutic Effect of Sophora Flavescens Ait, Ginger, Cornus Officinalis Compound On KK-Ay Diabetes Mellitus Mice | | |
Xiao-Fang Chen, Wei Wang, Chan Jia
Institute of Medicinal Plants Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
Correspondence: Xian-En Li e-mail: [email protected]
Objective: Diabetic nephropathy (DN) is one of the major micro-vascular complications of diabetes mellitus and is the most common cause of end-stage renal diseases. Chinese herbal compound has potential to decrease blood glucose and protect renal. To observe the effects of Chinese herbal compound (Sophora flavescens Ait :ginger : Cornus officinalis = 3:4:4, SGC for short) and their concentrations on KK-Ay diabetes mellitus mice so as to explore the therapeutic effect on diabetic nephropathy and their dose-dependent manners. Methods: Male KK-Ay mice randomly divided into diabetic model group, metformin group, low-, moderate- and high-dose groups treated by SGC, C57BL/6J mice as control group. Body weight, fasting blood glucose oral glucose tolerance test were detected; Serum TG, HDL-c, CRE, BUN, Urine ALB, CRE were detected by an automatic biochemical analyzer. Results: After treated SGC, the glucose tolerance ability were significantly enhanced, and the levels of fasting blood glucose were decreased significantly. The level of TG, U-ALB/U-Cr, serum CRE, BUN was reduced Compared with diabetic model group. The high-dose SGC has best treatment effect on reducing level of blood glucose, TG, and U-ALB/U-Cr , while enhancing level of HDL-c. Conclusion: Chinese herbal compound SGC have a significantly therapeutic effect on type 2 diabetes mellitus drugs and an added protective effect beyond their influence on diabetic nephropathy. | Figure 1: HDL-c, U-ALB/U-Cr, serum CRE and BUN level at the end of experiment (P<0.05)
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Keywords: Sophora flavescens Ait; ginger; Cornus officinalis; dose-dependent manners; diabetic nephropathy; therapeutic effect
Acknowledgements: Institute of Medicinal Plants Development, Chinese Academy of Medical Sciences and Peking Union Medical college, Xian-En Li (National Nature Fund 81274014).
| P-34: Authenticity Identification of Gastrodia Tuber (Tianma) by HPLC Fingerprint | | |
Jiajia Xiao a, Youcheng Lei a, b, Tingwen Lin a, Yue Ma a, Guanghua Lu a
a School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China
b Haisco Pharmaceutical Group Co., LTD., Chengdu 611130, China
Correspondence: Guanghua Lu, e-mail: [email protected]; [email protected]
Objective: To develop HPLC fingerprint of Gastrodia tuber (Tianma, the tuber of Gastrodia elata Bl.) for genuine identification. Methods: 38 samples Tianma samples including tuber, slice, powder and 9 species of counterfeits samples were collected from different cultivation areas and herbal markets in China. An HPLC condition was optimized to separate 11 main peaks on the chromatograms of Tianma. The HPLC chromatograms of all herbal samples were collected and compared the similarities. The common peaks were further identified on the HPLC chromatograms of Tianma samples by compared to reference substance, retention time and UV spectra. Results: The chromatograms of Tianma samples were similar with correlation coefficient more than 0.780 (n = 38). Their mean chromatogram was calculated and used as Tianma HPLC fingerprint, on which 7 peaks were chosen as characteristic peaks and assigned as gastrodin, p-hydroxybenzyl alcohol, parishin A, B, C, E and adenosine, respectively. However, the chromatograms of 9 species of counterfeits were significantly different from Tianma fingerprint. The correlation coefficient between Tianma HPLC fingerprint with counterfeits chromatograms was less than 0.042 (n = 9). The characteristic peaks of Tianma were not observed on the chromatograms of these counterfeits. Conclusions: The developed HPLC fingerprint of Tianma was characteristic and can be used to identify the true or false Tianma samples.
Acknowledgements: The Key Programme of Science and Technology of Sichuan Province, China (No. 2011NZ0058).
| P-35: Rapid Quantification of Paeoniflorin and Albiflorin in Red Peony Root Using Near-Infrared Spectroscopy | | |
Yue Ma a, Yongqiang Liu a, Qing Li a, Jiajia Xiao a, Fei Long a, Zhigang Xu b, Guanghua Lu a
a School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China
b Wansheng Agriculture Science and Technology Co. Ltd., Zhongjiang 618100, China
Correspondence: Guanghua Lu, [email protected]; [email protected]
Objective: To rapidly quantify the amount of bioactive compounds in Red Peony Root (the roots of Paeonia lactiflora Pall., RPR) by near-infrared spectroscopy (NIR) for quality assessment. Methods: A total of 65 RPR samples were collected from Sichuan, Zhejiang and Anhui of China, and then quantified the amounts of paeoniflorin and albiflorin by NIR and ultra high performance liquid chromatography (UHPLC). 53 RPR samples were chosen as the calibration set, while the remaining 12 RPR samples were used as external validation set. The calibration models of paeoniflorin and albiflorin were further developed by modified partial least squares (MPLS) using UHPLC analytical data as reference. The contents of paeoniflorin and albiflorin were predicted in unknown samples by calibration models. Results: The square errors of prediction (SEP) and R2 of calibration set calculated by MPLS models for paeoniflorin were 1.4372 and 0.9805, respectively. The data for albiflorin were 0.7843 and 0.9886, respectively. Moreover, the SEP and R2 of external validation set for paeoniflorin were 0.7145 and 0.9741, respectively; and for albiflorin were 0.6324, 0.9890, respectively. The amounts of the two bioactive compounds quantified by NIR and UHPLC were consistent. Conclusions: NIR can be accurately quantified the amounts of chemical marker in RPR and rapidly assess its quality.
Acknowledgements: Scientific Research Fund of Sichuan Provincial Education Department, China (No. 16ZA0113).
| P-36: Bioassay of Antiplatelet Aggregation Bioactivity in Chuanxiong Rhizome | | |
Yixin Yao, Fang Hua, Wan Li, Haofeng Jiang, Shanshan Gao, Guanghua Lu
School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China
Correspondence: Guanghua Lu, [email protected]; [email protected]
Objective: Chuanxiong (Szechwan Lovage Rhizome) is a widely used Chinese herb to activate blood and dissolve stasis. But its quality cannot be effectively assessed by the known chemical marker. So, the study is to develop a bioassay method to quantify the antiplatelet aggregation bioactivity (AAB) for quality assessment. Methods: Chuanxiong sample was extracted in water. The extract was quantified AAB by experiment in vitro. The platelet aggregating in platelet-rich plasma was induced by ADP. The ratio of platelet inhibition was chosen as AAB marker. Sodium ferulate was a reference. AAB in analytes was quantified by the amount reaction of parallel line analysis (2-2′) method. Results: The reliability test for quantifying AAB was passed through. The measured value was valid for both solidum ferulate and Chuanxiong extract. The extracts showed the significant AAB (P < 0.01). The correlation coefficient was 0.9934 (n = 4) between the amount of AAB for solidum ferulate (0.5 ~ 2 mg/mL) and their ratios of platelet inhibition. The RSD on the amount of AAB was 3.43% (n = 6) by six replicated tests with the confidence limit rate 23.90% (n = 6). AAB were estimated to be 1.515 ~ 3.183 U/g in five Chuanxiong samples collected from various cultivation areas. Conclusions: The developed method can accurately quantify AAB in Chuanxiong rhizome and assess its quality.
Acknowledgement: Science and Technology Programme in Administration of Traditional Chinese Medicine of Sichuan, China (No. 2010-08).
| P-37: Antifibrotic Activity of PHY906, Scutellariae Radix and Baicalein | | |
Shujun Zhou a, Jun Yuan b, Zhitao Liang b, Naima Saeed a,c, Yunxia Li d, Cheng Peng d, Jingzheng Song e, Peter J Hylands a, Yung-Chi Chengf, Zhongzhen Zhao b, Qihe Xua
a Centre for Integrative Chinese Medicine & Department of Renal Sciences, King’s College London, London, UK
b School of Chinese Medicine, Hong Kong Baptist University (HKBU), Hong Kong, China
c Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan
d Pharmacy College, Chengdu University of TCM (CDUTCM), Chengdu, Sichuan, China
e PuraPharm International (H.K.) Limited, Nanning, Guangxi, China
f Department of Pharmacology, Yale University School of Medicine, New Haven, CT, USA
Correspondence: Shujun ZHOU e-mail: [email protected]; Qihe XU, e-mail: [email protected]
Introduction: PHY906 (KD018) is a botanical medicine containing four herbal constituents for treating liver carcinoma, a common malignancy in patients with liver cirrhosis. As it contains a known antifibrotic herb, Scutellariae Radix (SR), we set out to compare the antifibrotic effect of PHY906, its 4 herbal constituents, different SR extracts and SR-derived flavonoids in an in-vitro model of TGF-β1-induced fibrosis. Materials and Methods: PHY906 and its four herbal constituents were characterised at Yale; SR was authenticated at PuraPharm as dried roots of Scutellaria baicalensis Georgi. The five major SR flavonoids (F5, i.e., baicalin, baicalein, wogonoside, wogonin and oroxylin A) and the NRK-49F fibroblast cell line were authenticated and quality-assured at King’s. SR aqueous and methanolic extracts were prepared and characterised at HKBU. The in-vitro antifibrotic activity of all herbal materials was examined at King’s by picro-Sirius red staining and spectrophotometric analysis, as well as hydroxyproline assay and immunoassays of fibrogenic markers. Results: PHY906 showed a mild but insignificant antifibrotic trend. Its SR aqueous extract component, but not the three other herbal components showed a mild but significant antifibrotic activity. SR methanolic extract was 8-fold more enriched in F5 (22.7% vs 2.8%) and had a 4-fold stronger antifibrotic activity (IC50 39.6±4.1 vs 155.6±10.3 μg/ml) than SR aqueous extract. The IC50 of F5 ranged from 38.2±4.7 μM (baicalein) to 190.0±11.2 μM (oroxylin A), P<0.05. Baicalein was also the least cytotoxic (P<0.05) among the F5. Conclusions: Despite the mild antifibrotic activity of its SR aqueous extract component, PHY906 is a poor antifibrotic. The SR methanolic extract containing more flavonoids shows more potent antifibrotic activity than the SR aqueous extract. Among the F5, baicalein shows the most potent antifibrotic activity and the least cytotoxicity. Thus, the antifibrotic activities of the SR methanolic extract and baicalein deserve to be further investigated.
Acknowledgements: This work was supported by a PuraPharm Scholarship award (QX, PJH), a HKBU Faculty Research Grant (ZZ, QX), a CDUTCM open fund (QX, PJH, CP) and a Sino-British Fellowship Trust Award (SZ).
| P-38: A New Method On Investigate the Chemical Constituents Which Have an Anti-Thrombin Effect By HPLC | | |
Jia-Yu Wang, Wen-Zhen Hou, Ya-Jie Wei, Shan-Shan Gao, Guang-Can Bai, An-Dong Wang, Chang-Hua Ma*
Analysis of TCM, Beijing University of Chinese Medicine, Beijing 100102, China
Correspondence: Chang-Hua Ma, e-mail: [email protected]
Objective: An in vitro anti-thrombin bioassay was developed to investigate the chemical constituents which have an anti-thrombin effect from the water soluble components of Salvia miltiorrhiza. Methods: Use Chromozym TH as a probe combined with ethyl acetate - Semi-micro extraction to measure p-Nitroaniline by HPLC. Results: The inactivation rate of thrombin by Sodium Danshensu, Salvianolic acid A and Salvianolic Acid B under a given set of conditions were 3.06%, 77.77% and 2.35%. In the Salvia water-soluble components, Salvianolic acid A has a direct inhibition of thrombin, while Sodium Danshensu and Salvianolic Acid B have no significant effect on thrombin. Conclusions: The method is sensitive and low consumption. It can eliminate the interference absorbed for the sample itself which can be used for screening single or multiple direct antithrombin active ingredient of herbal extract.
| P-39: A Study on the Effects of Myricetin on Endothelial Injury in a Cellular Model of Hyperhomocysteinaemia | | |
Antony Kam a,b, Valentina Razmovski-Naumovski a,c,d, Kong M. Li e, Srinivas Nammi c,f, Kelvin Chan a,c,g , George Q. Li a
a Faculty of Pharmacy, University of Sydney, NSW 2006, Australia
b School of Biological Sciences, Nanyang Technological University, 637551, Singapore
c National Institute of Complementary Medicine, Western Sydney University, NSW 2560, Australia
d South Western Sydney Clinical School, School of Medicine, University of New South Wales, NSW, 2052, Australia
e Discipline of Pharmacology, Sydney Medical School, University of Sydney, NSW 2006, Australia
f School of Science and Health, Western Sydney University, NSW 2560, Australia
g School of Pharmacy & Biomolecular Sciences, Liverpool John Moores Univerversity, L3 3AF, United Kingdom
Correspondence: George Q. Li. e-mail: [email protected]; K CHAN, e-mail: [email protected]
Objectives: An increase in homocysteine and adenosine levels results in S-adenosylhomocysteine accumulation, and sensitise TNF-induced cytotoxicity [1]. In the search of natural products with beneficial effects against vascular complications associated with hyperhomocysteinaemia [2],[3], our laboratory has identified a common flavonoid in herbal medicine, myricetin (3,5,7,3′,4′,5′-hexahydroxyflavone), with potential benefits against hyperhomocysteinaemia. The aim of this study was to investigate the effects of myricetin on S-adenosylhomocysteine accumulation, and its protective effects against endothelial injury induced by the combination of homocysteine, adenosine and TNF in human endothelial cells (EAhy926). Methods: Intracellular S-adenosylhomocysteine level was determined by UPLC. Cytotoxicity and apoptosis were assessed by MTT dye reduction assay, annexin V/PI assay, JC-1 assay and western blot. Results: UPLC analysis showed that myricetin attenuated intracellular S-adenosylhomocysteine accumulation induced by homocysteine and adenosine. Myricetin also exerted protective effects against TNF-induced endothelial cytotoxicity and apoptosis sensitised by the combination of homocysteine and adenosine. In addition, it restored the depleted protein expression of DNA methyltransferase 1 (DNMT1) caused by the combination of homocysteine, adenosine and TNF. Conclusions: Myricetin elicits protective effects against hyperhomocysteinaemia-associated endothelial injury, providing new insights into its potential effects in maintaining cellular methylation homeostasis.
| P-40: Protective Effects of Qidengmingmu Herbal Formulation in in vitro and in vivo Models of Vascular Disruption | | |
Yining Neo a, Fuwen Zhang b, Liwen Han c, Kechun Liu c, Jun-guo Duan b, Tai-Ping Fan a
a Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1PD, UK
b Key Laboratory for Visual Function and Ophthalmopathy, Chengdu University of Traditional Chinese Medicine, Chengdu 610032, Sichuan Province, China
c Biology Institute of Shandong Academy of Sciences, Jinan 250014, China
Correspondence: Tai-Ping Fan, e-mail: [email protected]
Objective: Current treatments of diabetic retinopathy (DR) are expensive, only help to slow down its progress and often accompanied with adverse side effects. Here, we examined the cellular mechanisms of the QiDengMingMu capsule, an oral herbal formulation undergoing extensive clinical studies in China as a potentially safe and affordable alternative. Methods: Human retinal microvascular endothelial cells (HRMECs) and human pericytes from placenta (hPC-PLs) were challenged with glucose, cobalt(II) chloride (CoCl2, a hypoxia-inducing agent), taxol (a tubulin targeting agent) and VEGF. Using these models of vascular disruption, QiDengMingMu formulation (QF), its three herbal crude extracts (Puerariae Lobatae Radix; Radix Astragali; Erigeron Breviscapus) as well as its three main active compounds were individually tested for their ability to (i) modulate viabilities of HRMECs and hPC-PLs under normal conditions or in presence of VEGF, (ii) protect these cells against damage induced by CoCl2 or taxol. The in vivo effects of QF and its components on vascular development were studied in a transgenic zebrafish embryo model. PTK787/ZK222584, a potent inhibitor of VEGF receptor tyrosine kinases, was used to inhibit the growth of intersegmental vessels (ISVs) of zebrafish embryos. Results: QF did not cause any cytotocity effect on HMRECs and hPC-PLs. High concentrations of QF were able to reverse inhibitory effects of CoCl2 in HMRECs. QF and its three individual herbal extracts were able to partially restore the growth of ISVs in a dose-dependent manner. Conclusions: QiDengMingMu capsule has exhibited its potential in angiogenesis modulation, hence possibly treating DR and other ocular disorders.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
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