|Year : 2022 | Volume
| Issue : 1 | Page : 92-99
Pharmacokinetic comparison of four major bio-active components of naoxintong capsule in normal and acute blood stasis rats using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry
Wei-Xia Li1, Shu-Qi Zhang2, Man-Man Li2, Hui Zhang2, Xiao-Yan Wang3, Lu Niu4, Jin-Fa Tang1, Xue-Lin Li1
1 Department of Pharmacy, Henan Province Engineering Laboratory for Clinical Evaluation Technology of Chinese Medicine, The First Affiliated Hospital of Henan University of Chinese Medicine; School of Pharmacy, Henan University of Chinese Medicine; Provincial and Ministerial co Construction Collaborative Innovation Center for Prevention and Treatment of Respiratory Diseases with Traditional Chinese Medicine of Henan University of Chinese Medicine, Zhengzhou, China
2 Department of Pharmacy, Henan Province Engineering Laboratory for Clinical Evaluation Technology of Chinese Medicine, The First Affiliated Hospital of Henan University of Chinese Medicine, bSchool of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, China
3 Department of Pharmacy, Henan Province Engineering Laboratory for Clinical Evaluation Technology of Chinese Medicine, The First Affiliated Hospital of Henan University of Chinese Medicine; Provincial and Ministerial co Construction Collaborative Innovation Center for Prevention and Treatment of Respiratory Diseases with Traditional Chinese Medicine of Henan University of Chinese Medicine, Zhengzhou, China
4 School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, China
|Date of Submission||02-Jul-2020|
|Date of Acceptance||18-Feb-2021|
|Date of Web Publication||29-Jan-2022|
Prof. Jin-Fa Tang
The First Affiliated Hospital of Henan University of Chinese Medicine, No. 19 Renmin Road, Jinshui District, 450008 Zhengzhou
Prof. Xue-Lin Li
The First Affiliated Hospital of Henan University of Chinese Medicine, No. 19 Renmin Road, Jinshui District, 450008 Zhengzhou
Source of Support: None, Conflict of Interest: None
Objective: To compare the pharmacokinetic differences of the main components of Naoxintong capsule (NXTC) in normal and acute blood stasis rats. Materials and Methods: Rats were subcutaneously injected with adrenaline hydrochloride twice; during the two subcutaneous injections, the rats were placed in ice water for 4 min to reproduce the model rat of acute blood stasis. The normal and acute blood stasis rats were administrated a 5.04 g/kg dose of NXTC suspension. Then, blood samples were collected from the posterior retinal venous plexus at different time points. Plasma concentrations of four major bio-active components including caffeic acid, ferulic acid, formononetin, and tanshinone IIA in NXTC were measured using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. Phoenix WinNonlin v6.2 software was used to calculate the pharmacokinetic parameters. Results: Compared with the normal rats, the acute blood stasis rats showed a significant decrease in Cmax of ferulic acid and formononetin, AUCall of caffeic acid and ferulic acid, and AUCINF_obs of ferulic acid. Conversely, an increase in the Vz_F_obs and MRTlast of ferulic acid and caffeic acid was observed. These findings demonstrate that the absorption of the four NXTC components was weakened in the acute blood stasis rats and that the elimination time was prolonged. Conclusions: The significant difference in some parameters of the four NXTC components between the normal and acute blood stasis rats might be caused by an increase in blood viscosity and the subsequent slowing down of blood flow in the acute blood stasis rats. The pharmacokinetic study conducted in pathological state can provide important information and scientific basis for further rational clinical application of NXTC.
Keywords: Acute blood stasis rat model; Naoxintong capsules; pharmacokinetics; ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry
|How to cite this article:|
Li WX, Zhang SQ, Li MM, Zhang H, Wang XY, Niu L, Tang JF, Li XL. Pharmacokinetic comparison of four major bio-active components of naoxintong capsule in normal and acute blood stasis rats using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. World J Tradit Chin Med 2022;8:92-9
|How to cite this URL:|
Li WX, Zhang SQ, Li MM, Zhang H, Wang XY, Niu L, Tang JF, Li XL. Pharmacokinetic comparison of four major bio-active components of naoxintong capsule in normal and acute blood stasis rats using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. World J Tradit Chin Med [serial online] 2022 [cited 2022 May 18];8:92-9. Available from: https://www.wjtcm.net/text.asp?2022/8/1/92/336835
| Introduction|| |
Blood stasis syndrome is one of the most common syndrome types in traditional Chinese medicine (TCM) syndrome differentiation, which can be involved in almost all clinical diseases, especially widely exists in cardio- and cerebrovascular diseases. Activating blood circulation and removing blood stasis (RBS) is the main treatment principle of blood stasis syndrome. Pharmacological studies have confirmed that the TCM principle of promoting blood circulation and RBS has imparted certain effects, including improving hemorheology, platelet function, and hemodynamics; protecting vascular endothelial cells; and anticoagulation.
Naoxintong capsule (NXTC) is a classic prescription for “treating both brain and heart” theory. It is prepared using Buyang Huanwu Decoction, a traditional Chinese prescription recorded in the ancient medical book Yi-Lin-Gai-Cuo written by Qingren Wang of Qing Dynasty. NXTC is composed of 16 traditional Chinese herbs (TCHs), including 11 plant herbs (Huangqi, Chishao, Danshen, Danggui wei, Chuanxiong, Taoren, Honghua, Jixueteng, Niuxi, Guizhi, and Sangzhi), 2 resin herbs (Ruxiang and Moyao), and 3 animal herbs (Dilong, Quanxie, and Shuizhi). NXTC is a one of the commercial medicinal products approved by the China Food and Drug Administration and listed in the Chinese Pharmacopoeia 2015. NXTC possess the effect of invigorating qi and improving blood circulation, RBS, and dredging collaterals. Modern research has shown that the pharmacological effects of NXTC mainly focus on anti-myocardial ischemia and ischemia/reperfusion (I/R) injury, anti-atherosclerosis, anti-myocardial fibrosis, anti-cerebral I/R injury, learning improvement, memory functions, etc. Hence, NXTC is commonly used for the treatment of coronary heart disease, stroke, angina pectoris, cerebral infarction, secondary prevention of myocardial ischemia, transient ischemic attack, carotid atherosclerosis, vertebro-basilar insufficiency, and other cardio- and cerebrovascular diseases.
An increasing number of studies show that the pharmacokinetic parameters of TCH components are altered in morbidity body and significantly differ from those in normal states., Therefore, the alterations in pharmacokinetic parameters under pathological conditions should be studied, the data of which could be more beneficial for clinical applications than those obtained in normal conditions. Several studies have detailed the pharmacokinetics and metabolites of the main components in NXTC. The studies demonstrated that there were 36 prototype compounds and 52 metabolites of NXTC identified or tentatively characterized in Beagle dog urine and feces; the pharmacokinetic profiles of caffeic acid, ferulic acid, formononetin, cryptotanshinone, and tanshinone IIA in healthy rats have already been reported.
Based on this, we employed the pharmacokinetic comparison of the main active components of NXTC between normal and blood stasis model rats. The study will provide valuable data that will highlight the differences in the mechanism of NXTC for invigorating qi, promoting blood circulation, RBS, and dredging collaterals after oral administration. The study results will also improve the clinical applications of NXTC.
| Materials and Methods|| |
Reagents and materials
NXTCs (Shanxi Buchang Pharmaceutical Co., Ltd., batch number: 190193) and adrenaline hydrochloride injection (Suicheng Pharmaceutical Co., Ltd., specifications: 1 mL: 1 mg, batch number: 61903011) were purchased from pharmacy of the First Affiliated Hospital of Henan University of Chinese Medicine (Zhengzhou, China). Reference standards of caffeic acid, ferulic acid, formononetin, and tanshinone II A were purchased from Nanjing Liangwei Biological Technology Co., Ltd. with purity above 98% (Nanjing, China). Internal standards (IS) including clarithromycin and probenecid were purchased from the National Institutes for Food and Drug Control with purity above 98% (Beijing, China). Molecular water purification system was used for ultra-pure water purification. Acetonitrile and methanol (HPLC grade) were supplied by Merck KGaA (Darmstadt, Germany). HPLC-grade formic acid was purchased from Dikma Technology Co., Ltd (Tianjin, China).
Naoxintong capsule solution preparation
The required content of NXTC was dispersed in 0.5% CMC-Na aqueous solution to prepare a NXTC suspension with a concentration of 0.33 g/mL.
Preparation of standard stock solutions and quality control samples
Chemical structures of the four components in NXTC are shown in [Figure 1]. The mixture of standard stock solution was prepared by dissolving an accurately weighed quantity of caffeic acid, ferulic acid, formononetin, and tanshinone II A in methanol and had a concentration of 30.5, 30, 38, and 21.5 μg/mL, respectively. Standard working solutions were provided by the stock solution serially diluted with methanol. The solutions prepared had a concentration of 0.5, 1, 5, 10, 50, and 100 ng/mL for caffeic acid; 1, 5, 10, 50, 100, and 500 ng/mL for ferulic acid; 0.05, 0.1, 0.5, 1, 5, and 10 ng/mL for formononetin; and 0.5, 1, 5, 10, 50, and 100 ng/mL for tanshinone II A.
The IS stock solutions of clarithromycin and probenecid that were prepared had a concentration of 532 μg/mL and 536 μg/mL, respectively. Further, the mixture of IS working solution was diluted with methanol, and the final concentrations for clarithromycin and probenecid were 106.4 ng/mL and 107.2 ng/mL, respectively.
QC samples were prepared at concentrations of 30, 50, and 500 ng/mL for caffeic acid; 10, 20 and 200 ng/mL for ferulic acid; 0.5, 1, and 10 ng/mL for formononetin; and 1, 5, and 50 ng/mL for tanshinone II A. All solutions were stored at 4°C until the time for analysis.
Conditions of chromatography and mass spectrometry
Thermo Scientific™ TSQ Altis™ was used for liquid chromatographic analysis and mass spectrometry detection. Acquity ultra-performance liquid chromatography coupled BEH C18 column (2.1 mm × 100 mm, 1.7 μm) was carried out for chromatographic separation maintained at 30°C. The gradient conditions of mobile phase were 0.1% aqueous formic acid (A) and methanol (B): at 0–2 min, 40%–100% B; at 2–5 min, 100% B; at 5–5.1 min, 100%–40% B; and at 5.1–8 min, 40% B. The flow rate was set at 0.2 mL/min. The temperature of autosampler was conditioned at 10°C and the injection volume was 2 μL.
The positive and negative ESI ionization modes were used for data acquisition. The parameters in the ESI source were set as follows: positive ion scanning voltage, 3500 V; negative ion scanning voltage, 2500 V; sheath gas velocity, 25 arb; auxiliary air velocity, 7 arb; collision gas pressure, 1.5 mTorr; ion transmission tube temperature, 325°C; and ion source temperature, 400°C. The multiple reaction monitoring (MRM) mode was used for scanning mode. The selected monitor ions were m/z 178.96–135.04 for caffeic acid, m/z 192.95–134.00 for ferulic acid, m/z 267.00–251.97 for formononetin, m/z 295.05–277.20 for tanshinone IIA, m/z 748.35–590.29 for clarithromycin (IS1), and m/z 284.00–240.05 for probenecid (IS2). The optimized collision energy for precursor/product ion pairs of the 6 analytes was set as 15.88, 14.7, 20.27, 19.78, 17.7, and 14.97 eV, respectively, and the RF-lens were set as 49, 44, 68, 67, 74, and 57 V, respectively.
Female Sprague–Dawley (SD) rats (6–8 weeks old, weighing 220 ± 20 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd (Beijing, China). Prior to the experiment, the rats were fed adaptively for 1 week. The circadian rhythm was maintained for 12 h, and the rats were allowed to drink and feed freely. The temperature and humidity met the requirements of animal feeding. On the basis of “Detailed Rules and Regulations for Administration and Implementation of Biomedical Animal Experiments” (No. 1998-55, Ministry of Public Health, China), animal care and experimental protocols were performed. The study protocol was approved by the Ethical Committee of the First Affiliated Hospital of Henan University of Chinese Medicine.
The model rats of acute blood stasis were established by ice-bath swimming and adrenaline hydrochloride injection. Adrenaline hydrochloride injection at a dose of 0.8 mg/kg was used for subcutaneous injection.,, Two hours after the first subcutaneous injection, the rats were placed in 0°C–2°C ice-cold water for 4 min, and then 2 h later, a second subcutaneous injection with 0.8 mg/kg adrenaline hydrochloride injection was given.
There were ten SD rats in each of the normal group and the model group. After 7 days of acclimatization, all rats were fasted and kept in water for 12 h prior to the experiment. Then, rats of normal and model groups were orally administrated with 5.04 g/kg of the NXTC suspension. At different time points (0, 0.083, 0.167, 0.25, 0.5, 0.75, 1, 1.5, 2, 4, 6, 8, 10, and 12 h), blood samples were collected from the eye orbital venous plexus of the rats after administration into the heparinized centrifuge tube.
The blood samples were centrifuged at 3000 rpm for 10 min at 4°C to obtain the plasma samples, which were stored at − 80°C until determination. Briefly, 20 μL 10% formic acid water, 10 μL IS solution, and 300 μL methanol were added to 100-μL plasma samples. The mixture was vortexed for 3 min and centrifugated at 14,000 rpm for 10 min at 4°C. The supernatant was transferred into a new centrifuge tube and concentrated until dry under vacuum condition. The residue was re-dissolved in 100 μL methanol, vortexed for 3 min, and centrifugated at 14,000 rpm for 10 min. The supernatant was then obtained for analysis.
Reference standard and IS solution both were added into blank plasma samples from normal rats, and plasma samples after administration of NXTC were used for the specificity analysis. By comparing blank plasma samples from the normal rats and blood stasis rats, the specificity could be calculated.
Linearity, limit of detection, and lower limit of quantification
By plotting the peak area ratio (y) of the analytes to the corresponding IS versus the nominal concentration (x) of analytes with weighted (1/x2) least square linear regression, the linearity of each calibration curve was determined. The signal-to-noise ratio (S/N) of 3:1 was determined as limit of detection (LOD), and the lower limit of quantification (LLOQ) with S/N ratio of 10:1 was determined as the lowest concentration.
Accuracy and precision
Three concentration levels of QC samples (n = 6) were used on the same day and three consecutive validation days to evaluate the inter- and intra-day accuracy and precision of the method, respectively. Relative percentage error and relative standard deviation (RSD%) were used for the expression of accuracy and precision, respectively.
Extraction recovery and matrix effect
The ratio of the mean concentration between the final QC samples against those blank plasma samples initially added with analytes was used to calculate the extraction recovery.
By comparing the peak areas of postextraction plasma samples spiked with standard solutions to those obtained from the neat standard solutions at equivalent concentrations, the matrix effect was calculated.
Six sets of low, middle, and high concentrations of QC samples under different temperature conditions were used to investigate the stability. QC samples were placed in refrigerated conditions (4°C) for 24 h to evaluate the short-term stability. The freeze-thaw cycle stability was assessed by freezing the samples at −80°C and thawing them at room temperature for three cycles with six duplicates.
Data processing and statistics
The data were collected and analyzed by Tracefinder 4.1 software (Thermo Fisher Scientific, Waltham, USA). The major pharmacokinetic parameters were processed and calculated by Phoenix Winnonlin 6.2 software (Pharsight Corporation, USA) using noncompartment model. SPSS 20.0 was used for statistical analysis and an independent-sample t-test was used for comparison. The experiment data were presented as mean ± SD. P <0.05 indicated that there was a statistically significant difference.
| Results|| |
Specificity and linearity
No significant interference was found among endogenous components, four analytes, and ISs at the retention times. The representative MRM chromatograms are shown in [Figure 2]. The calibration curve, correlation coefficient, LOD, and LLOQ for caffeic acid, ferulic acid, formononetin, and tanshinone IIA in NXTC are shown in [Table 1].
|Figure 2: Typical multiple reaction monitoring chromatograms of Naoxintong capsule components in rat plasma|
Click here to view
|Table 1: Calibration curve, limit of detection, and lower limit of quantification for four components of Naoxintong capsule in rat plasma samples|
Click here to view
Precision and accuracy
The inter-/intra-day precision and accuracy of QC samples are presented in [Table 2]. The method showed good precision and accuracy, and all of the results were within the accepted variable limits.
|Table 2: Precision and accuracy of Naoxintong capsule constituents in rat plasma samples|
Click here to view
Recovery and matrix effect
Extraction recovery and matrix effect of the four components in NXTC are shown in [Table 3]. The extraction recovery of the four components was observed to be between 69.09% and 97.20% (RSD ≤15.94%) at three concentrations of analytes. The matrix effect ranged from 71.82% to 119.00% (RSD <15%).
|Table 3: Recovery and matrix effect of Naoxintong capsule constituents in rat plasma samples|
Click here to view
The results indicated that the four analytes of NXTC in rat plasma were stable under different conditions. The results are shown in [Table 4].
|Table 4: The stability of four Naoxintong capsule constituents in rat plasma samples|
Click here to view
Contents of four components in Naoxintong capsule
The contents of caffeic acid, ferulic acid, formononetin, and tanshinone IIA were determined to be 0.011, 0.099, 0.060, and 0.148 mg/g, respectively.
Pharmacokinetic comparison under normal and blood stasis conditions
The mean plasma concentration-time profiles of caffeic acid, ferulic acid, formonetin, and tanshinone IIA in NXTC in the normal and acute blood stasis model rats are shown in [Figure 3]. The partial pharmacokinetic parameters of these four analytes are presented in [Table 5]. Compared with those of the normal rats, the values of peak time (Tmax) of the four components in the acute blood stasis model rats were prolonged with no remarkable differences; however, a decrease was observed in the values of peak concentrations (Cmax), the difference in ferulic acid and formononetin was statistically significant (P < 0.05); AUCall values of caffeic acid and ferulic acid statistically significantly reduced (P < 0.05). Further, a statistically significant decrease was observed in the AUCINF_obs values of ferulic acid (P < 0.01). Conversely, a statistically significant increase was observed in the Vz_F_obs and the mean retention time (MRTlast) of caffeic acid and ferulic acid (P < 0.05). The results suggest that the blood circulation disorder caused by acute blood stasis hinders the absorption of the four components. The difference in the above pharmacokinetic parameters might be attributed to the characteristics of “viscosity, concentration, coagulation, and aggregation” of the blood in the acute blood stasis model rats, that is, increased blood viscosity, slowed blood flow and absorption of components, and prolonged Tmax and MRTlast.
|Figure 3: Blood concentration–time curves of caffeic acid (a), ferulic acid (b), formononetin (c), and tanshinone IIA (d) in normal and acute blood stasis model rats (normal group: n = 7, model group: n = 6)|
Click here to view
|Table 5: Pharmacokinetic parameters of four components in Naoxintong capsule between normal and acute blood stasis model rats|
Click here to view
| Discussion|| |
Blood stasis syndrome is a pathological state of blood circulation, which is described as slowing of blood flow or accumulation of blood due to xinqi disorder in TCM. TCM believes that the “sadness” of seven emotions (qiqing) and the “cold evil” of six evils (liuxie) are the main causes of acute blood stasis. Currently, blood stasis is usually understood as a blood system disease, and pathological studies have shown that blood stasis is mainly characterized by cardio- and cerebrovascular diseases, such as cerebral infarction, myocardial infarction, coronary heart disease, and hypertension.,, The pharmacologic effects of NXTC involve replenishing qi and activating blood, RBS, and dredging collaterals. NXTC is often used in the treatment of cardio- and cerebrovascular diseases, which is closely related to its main components of caffeic acid, ferulic acid, formononetin, and tanshinone IIA. Numerous studies have demonstrated that caffeic acid and its derivatives have pharmacological effects such as antioxidation, immune regulation, anti-cancer, regulation of cardiovascular and cerebrovascular diseases, and protection of brain tissue damage.,, Ferulic acid is a metabolite of caffeic acid methylation, which can promote bone marrow hematopoiesis, enhance immunosuppression, protect cardiovascular system, reduce blood lipid, resist arteriosclerosis, and inhibit platelet aggregation.,, Formononetin exerts pharmacological effects, including improving atherosclerosis and inhibiting the proliferation of vascular smooth muscle cells., In recent years, tanshinone IIA has attracted the attention of researchers for use in cardiovascular and cerebrovascular disorders. The protective effects of tanshinone IIA on the heart include preventing the formation of atherosclerosis, preventing myocardial injury and hypertrophy, expanding coronary arteries, and related mechanisms of action.,,,
The pharmacokinetic parameters of a drug in a pathological state are different from those in a normal condition owing to changes in the physiological state and biochemical response of the body. Given that NXTC is usually used for the treatment of patients with blood stasis, we studied the pharmacokinetics of NXTC in acute blood stasis rats. Under acute blood stasis condition, an increase was observed in factors such as blood viscosity, coagulation degree, blood aggregation, and vascular obstruction because of the abnormal hemorheology., In the model group, the Tmax and Cmax of the four components were prolonged and decreased, respectively. A change was observed in parameters including AUCall, Vz_F_obs, and MRTlast, indicating that the absorption and metabolism of NXTC were affected in acute blood stasis. These results provide important information for guiding the clinical rational use of NXTC.
| Conclusions|| |
Herein, we have established the analytical method to probe the pharmacokinetic properties of the four main ingredients – caffeic acid, ferulic acid, formononetin, and tanshinone IIA – of NXTC in acute blood stasis rats in comparison with the corresponding profiles in normal rats. The parameters demonstrated a significant decrease in the Cmax of ferulic acid and formononetin and a significant decrease in the AUCall/AUCINF_obs of ferulic acid and the AUCall of caffeic acid. An increase was observed in the plasma concentration and MRTlast of ferulic acid and caffeic acid. The pharmacokinetic alterations were consistent with the changes in rheological characteristics of the acute blood stasis rats. These investigations may provide experimental basis for tailoring the clinical application of NXTC to patients with blood stasis.
Financial support and sponsorship
This work was supported by the National Science and Technology Major Project of China “Key New Drug Creation and Manufacturing Program” 2015ZX09501004-001-007, National Natural Science Foundation of China 82004082, and Top talent training project of TCM in Henan Province.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Xian MH, Ji SL, Chen C, Liang SW, Wang SM. Sparganin A alleviates blood stasis syndrome and its key targets by molecular docking. RSC Adv 2019;9:37978-85.
Guo JW, Liu MJ. Meta-analysis of acute intracerebral hemorrhage treated with traditional Chinese medicine or/and composition of promoting blood circulation and removing blood stasis. J China Japan Friendship Hosp 2001;15:283-6.
Zhao BC, Zhao MZ. Prevention and treatment strategy of cardiovascular and cerebrovascular diseases guided by the theory of “concurrent treatment of the brain and heart”. Chin J Integr Tradit West Med 2013;33:1596-8.
Wang H, Qiu L, Ma Y, Zhang L, Chen L, Li C, et al.
Naoxintong inhibits myocardial infarction injury by VEGF/eNOS signaling-mediated neovascularization. J Ethnopharmacol 2017;209:13-23.
Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China. Beijing: China Medical Science Press; 2015. p. 1379-80.
Long-Tao L. Chinese experts consensus on clinical application of naoxintong capsule. Chin J Integr Med 2018;24:232-6.
Han J, Tan H, Duan Y, Chen Y, Zhu Y, Zhao B, et al.
The cardioprotective properties and the involved mechanisms of NaoXinTong Capsule. Pharmacol Res 2019;141:409-17.
Pfeifer S. The effect of disease on the pharmacokinetics of drugs. 3. Cardiovascular, respiratory, neoplastic, thyroid diseases, diabetes, infections, mucoviscidosis, obesity, operations, burns etc., Pharmazie 1991;46:830-9.
Yan R, Yang Y, Chen Y. Pharmacokinetics of Chinese medicines: Strategies and perspectives. Chin Med 2018;13:24.
Li W, Guo J, Tang Y, Wang H, Huang M, Qian D, et al.
Pharmacokinetic comparison of ferulic acid in normal and blood deficiency rats after oral administration of Angelica sinensis, Ligusticum chuanxiong and their combination. Int J Mol Sci 2012;13:3583-97.
He Y, Su W, Chen T, Zeng X, Yan Z, Guo J, et al.
Identification of prototype compounds and derived metabolites of naoxintong capsule in beagle dog urine and feces by UFLC-Q-TOF-MS/MS. J Pharm Biomed Anal 2019;176:112806.
Li J, Bai Y, Bai Y, Zhu R, Liu W, Cao J, et al.
Pharmacokinetics of caffeic acid, ferulic acid, formononetin, cryptotanshinone, and tanshinone IIA after oral administration of Naoxintong capsule in rat by HPLC-MS/MS. Evid Based Complement Alternat Med 2017;2017:9057238.
Li WX, Huang MY, Tang YP, Guo JM, Shang EX, Liu X, et al.
Establishment and optimization of acute blood stasis rat model. Chin Pharm Bull 2011;27:1761-5.
Shen D, Ma N, Yang Y, Liu X, Qin Z, Li S, et al.
UPLC-Q-TOF/MS-based plasma metabolomics to evaluate the effects of aspirin eugenol ester on blood stasis in rats. Molecules 2019;24:1-18.
Liu L, Duan JA, Tang Y, Guo J, Yang N, Ma H, et al.
Taoren-Honghua herb pair and its main components promoting blood circulation through influencing on hemorheology, plasma coagulation and platelet aggregation. J Ethnopharmacol 2012;139:381-7.
Chiu CC, Lan CY, Chang YH. Objective assessment of blood stasis using computerized inspection of sublingual veins. Comput Methods Programs Biomed 2002;69:1-12.
Xue M, Chen KJ, Yin HJ. Relationship between platelet activation related factors and polymorphism of related genes in patients with coronary heart disease of blood-stasis syndrome. Chin J Integr Med 2008;14:267-73.
Yue SJ, Xin LT, Fan YC, Li SJ, Tang YP, Duan JA, et al.
Herb pair Danggui-Honghua: Mechanisms underlying blood stasis syndrome by system pharmacology approach. Sci Rep 2017;7:40318.
Li HX, Han SY, Wang XW, Ma X, Zhang K, Wang L, et al.
Effect of the carthamins yellow from Carthamus tinctorius
L. on hemorheological disorders of blood stasis in rats. Food Chem Toxicol 2009;47:1797-802.
Yao KW, Wang J, Zhu CL, Wu JT, Fang JZ. Results of different quantitative diagnosis analysis on the symptoms and signs of blood stasis syndrome in coronary heart disease. World Sci Technol 2009;11:684-8.
Tsai SJ, Chao CY, Yin MC. Preventive and therapeutic effects of caffeic acid against inflammatory injury in striatum of MPTP-treated mice. Eur J Pharmacol 2011;670:441-7.
Anwar J, Spanevello RM, Thomé G, Stefanello N, Schmatz R, Gutierres J, et al.
Effects of caffeic acid on behavioral parameters and on the activity of acetyl-cholinesterase in different tissues from adult rats. Pharmacol Biochem Behav 2012;103:386-94.
Kumaran KS, Prince PS. Caffeic acid protects rat heart mi-tochondria against isoproterenol-induced oxidative damage. Cell Stress Chaperon 2010;15:791-806.
Colombo R, Papetti A. An outlook on the role of decaffeinated coffee in neurodegenerative diseases. Crit Rev Food Sci Nutr 2020;60:760-79.
Chaudhary A, Jaswal VS, Choudhary S, Sonika, Sharma A, Beniwal V, et al.
Ferulic acid: A Promising therapeutic phytochemical and recent patents advances. Recent Pat Inflamm Allergy Drug Discov 2019;13:115-23.
Li Y, Cao SY, Lin SJ, Zhang JR, Gan RY, Li HB. Polyphenolic profile and antioxidant capacity of extracts from Gordonia axillaris
fruits. Antioxidants (Basel) 2019;8:1-14.
Zhang X, Wang Q, Wang X, Chen X, Shao M, Zhang Q, et al.
Tanshinone IIA protects against heart failure post-myocardial infarction via AMPKs/mTOR-dependent autophagy pathway. Biomed Pharmacother 2019;112:108599.
Wang H, Zhong L, Mi S, Song N, Zhang W, Zhong M. Tanshinone IIA prevents platelet activation and down-regulates CD36 and MKK4/JNK2 signaling pathway. BMC Cardiovasc Disord 2020;20:81.
Lu J, Shan J, Liu N, Ding Y, Wang P. Tanshinone IIA can inhibit angiotensin II-induced proliferation and autophagy of vascular smooth muscle cells via regulating the MAPK signaling pathway. Biol Pharm Bull 2019;42:1783-8.
Song T, Yao Y, Wang T, Huang H, Xia H. Tanshinone IIA ameliorates apoptosis of myocardiocytes by up-regulation of miR-133 and suppression of Caspase-9. Eur J Pharmacol 2017;815:343-50.
Braun H, Bueche CZ, Garz C, Oldag A, Heinze HJ, Goertler M, et al.
Stases are associated with blood-brain barrier damage and a restricted activation of coagulation in SHRSP. J Neurol Sci 2012;322:71-6.
[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]