Simultaneous determination of 10 components in Guanxinshutong capsules by quantitative analysis of multi-components by single marker / 药学学报

This study establishes a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of gallic acid, sodium danshensu, protocatechuic acid, protocatechuic aldehyde, vanillin, rosmarinic acid, salvianolic acid B, eugenol, cryptotanshinone and tanshinone IIA in Guanxinshutong capsules (Bambusae Concretio Silicea, Salvia miltiorrhiza, clove, borneol, Bambusae Concretio Silicea) by HPLC. Sample was loaded onto an Agilent C18 (ZORBAX Extend-RP C18, 250 mm × 4.6 mm, 5 µm) column and eluted with methanol-0.4% aqueous formic acid solution as a flow phase gradient, flow speed 1.0 mL·min-1, detection wavelength 280 nm, column temperature 35 ℃ and sample intake of 5 µL. Using protocatechuic acid as the internal reference, a relative correction factor was calculated and the durability was investigated, and the content of 10 components was calculated by QAMS and external standard method (ESM). The results show that the specificity, linear relationship, precision, repeatability, and stability of the 10 components were good. The average recovery was 98.20%-103.47% and RSD was 1.26%-2.84%. The relative positive factors and contents of the other nine components were calculated as gallic acid (0.759, 227.381), sodium tanshinol (3.630, 3.283), protocatechualdehyde (0.185, 0.150), vanillin (0.532, 65.213), rosmarinic acid (4.240, 1.035), salvianolic acid B (3.245, 18.204), eugenol (1.729, 9.265), cryptotanshinone (0.691, 1.449), and tanshinone ⅡA (0.702, 1.939). The results of QAMS were consistent with ESM analysis, and the relative error was between -3% and 3%. This method is stable and reliable, and can be used for the determination of 10 components in Guanxinshutong capsules.

Exploration of “Component-Target-Pathway” of Guanxinshutong Caspules in Treatment of Coronary Heart Disease / 中国药学杂志

OBJECTIVE: To explore the"multi-component, multi-target, multi-pathway" network regulation mechanism of Guanxinshutong caspules (GXST) in treatment of coronary heart disease by network pharmacology method. METHODS: The chemical components of GXST were identified based on liquid chromatography-mass spectrometry (LC-MS), and the oral bioavailability (OB)≥30% and drug likeness (DL)≥0.18 were used as the screening conditions for obtaining active molecular compounds. The targets related to the active compounds were predicted through the Traditional Chinese Medicine Systems Pharmacology database (TCMSP).The relevant targets of coronary heart disease were searched through literature mining and multiple databases and compared with the predicted component targets. Finally, the targets were introduced into the Molecule Annotation System 3.0 (MAS 3.0)to analyze the main biological pathways. RESULTS: Forty-three compounds were identified in GXST based on LC-MS technology and 10 main active compounds were determined through OB and DL screening, such as ellagic acid, cryptotanshinone, and tanshinone. These components affected 26 key targets, such as CLP, LDLR, TNF, et al, and 49 KEGG pathways were involved (P<0.05). CONCLUSION: GXST produces therapeutic effect for coronary heart disease mainly through the TGF-beta, T cell receptors, and MAPK signaling pathway.This study further reveals the characteristics of the multi-component, multi-target and multi-pathway of GXST, and lays a certain foundation for further elucidation of the mechanism of GXST in the treatment of coronary heart disease.

Screening and analysis of key active constituents in Guanxinshutong capsule using mass spectrum and integrative network pharmacology / 中国天然药物

Guanxinshutong capsule (GXSTC) is an effective and safe traditional Chinese medicine used in the treatment of cardiovascular diseases (CVDs) for many years. However, the targets of this herbal formula and the underlying molecular mechanisms of action involved in the treatment of CVDs are still unclear. In the present study, we used a systems pharmacology approach to identify the active ingredients of GXSTC and their corresponding targets in the calcium signaling pathway with respect to the treatment of CVDs. This method integrated chromatographic techniques, prediction of absorption, distribution, metabolism, and excretion, analysis using Kyoto Encyclopedia of Genes and Genomes, network construction, and pharmacological experiments. 12 active compounds and 33 targets were found to have a role in the treatment of CVDs, and four main active ingredients, including protocatechuic acid, cryptotanshinone, eugenol, and borneol were selected to verify the effect of (GXSTC) on calcium signaling system in cardiomyocyte injury induced by hypoxia and reoxygenation. The results from the present study revealed the active components and targets of GXSTC in the treatment of CVDs, providing a new perspective to enhance the understanding of the role of the calcium signaling pathway in the therapeutic effect of GXSTC.

Screening and analysis of key active constituents in Guanxinshutong capsule using mass spectrum and integrative network pharmacology / 中国天然药物

Guanxinshutong capsule (GXSTC) is an effective and safe traditional Chinese medicine used in the treatment of cardiovascular diseases (CVDs) for many years. However, the targets of this herbal formula and the underlying molecular mechanisms of action involved in the treatment of CVDs are still unclear. In the present study, we used a systems pharmacology approach to identify the active ingredients of GXSTC and their corresponding targets in the calcium signaling pathway with respect to the treatment of CVDs. This method integrated chromatographic techniques, prediction of absorption, distribution, metabolism, and excretion, analysis using Kyoto Encyclopedia of Genes and Genomes, network construction, and pharmacological experiments. 12 active compounds and 33 targets were found to have a role in the treatment of CVDs, and four main active ingredients, including protocatechuic acid, cryptotanshinone, eugenol, and borneol were selected to verify the effect of (GXSTC) on calcium signaling system in cardiomyocyte injury induced by hypoxia and reoxygenation. The results from the present study revealed the active components and targets of GXSTC in the treatment of CVDs, providing a new perspective to enhance the understanding of the role of the calcium signaling pathway in the therapeutic effect of GXSTC.

Effect and mechanism of Guanxin-Shutongcapsule in the treatment of arterial elasticity on the patients with hypertension / 国际中医中药杂志

Objective To observe the effect ofGuanxin-Shutongcapsule in the treatment of arterial elasticity on patients with hypertension.Methods The hypertension patients who met the inclusion criteria were divided into treatment group (50 cases) and control group (52 cases). The control group was treated with antihypertensive drugs to control blood pressure within the normal range. The treatment group was treated withGuanxin-Shutongcapsule on the basis of the control group. All were given 8 weeks treatment. The main artery elastic parameters were meansured by the carotid-femoral pulse wave velocity (C-FPWV) and cervical-dorsal arterial pulse wave velocity (C-DPWV). The immune turbidimetric method was employed to enhance for the determination of high sensitive C reactive protein (hs-CRP); and radioimmunoassay was used to assess the serum IL-6, TNF-a, triglyceride (TG) and total cholesterol (TC). The blood pressure was monitored during the treatment.Results After the treatment, the level of hs-CRP (2.83 ± 1.35 mg/Lvs. 3.65 ± 1.38 mg/L,t=6.357), TNF-α (0.16 ± 0.08 mg/Lvs. 0.28 ± 0.07 mg/L,t=18.213), C-FPWV (13.85 ± 1.86 m/svs. 15.34 ± 1.78 m/s,t=6.524), C-DPWV (11.98 ± 1.45 m/svs. 12.87 ± 1.48 m/s,t=7.152) in treatment group was significantly lower than those in the control group (P<0.01).ConclusionGuanxin-Shutong capsule by inhibiting systemic inflammation, reducing and reversing atherosclerosis, and improving the arterial elasticity and blood pressure.

Effect of Guanxinshutong on cerebral ischemia reperfusion in rats learning and memory ability and brain Bcl-2,Bax protein expression / 中国免疫学杂志

Objective:Guanxinshutong on cerebral ischemia reperfusion ( I/R) in rats learning and memory ability and brain Bcl-2,Bax protein expression to explore the protection mechanisms of Guanxinshutong on cerebral I /R injury in brain tissue.Methods:Healthy Wistar rats for the study were randomly divided into sham operation group ,I/R model group,high dose Guanxinshutong (1 g/kg) and low dose Guanxinshutong (0.5 g/kg) group,n=15.cerebral I/R model was estabhished by using the left commom cartid artery ligation method,modeling each group were fed daily before saline ,saline,Guanxinshutong (1 g/kg) and Guanxinshutong (0.5 g/kg), continuous 7 days;3 d after water maze training while continuing administration ,continuous 7 d,the eight day ,Morris water maze test;after the test,the rats were sacrificed blood and brain tissue ,serum by ELISA of Bcl-2 and Bax protein levels change;using HE staining of rat brain pathology;using immunohistochemistry assay in rat brain organization of Bcl-2 and Bax protein levels changes.Results:Compared with the sham group ,Guanxinshutong high-dose group test latency slightly longer timers after platform and platform quadrant dwell decreased slightly ,serum and brain tissue slightly elevated levels of Bax protein ,Bcl-2 protein levels decreased slightly but not significantly different ( P>0.05 ) ,low-dose group Guanxinshutong through latency test in rats ,after the number of platforms and platform quadrant dwell time decreased , serum and brain tissue elevated levels of Bax protein , Bcl-2 protein content decreased significantly different(P<0.01),cerebral I/R model group was significantly prolonged latency test ,after the number of platforms and platform quadrant dwell time significantly reduced serum and brain tissue was significantly increased Bax protein ,Bcl-2 protein content decreased with significant differences ( P<0.01 ).Conclusion: Guanxinshutong can significantly improve spatial memory , and by reducing Bax protein content and increased Bcl-2 protein,inhibition of cerebral I/R injury of apoptosis,and thus the brain tissue I/R has a protective effect.

Effect of Guanxin-Shutong capsule on AT1、ERK2 expression in rats with chronic heart failure / 国际中医中药杂志

Objective By investigating the effects of AT1 and ERK2 signaling pathway in CHF,to study the effect of Guanxin-Shutong capsule on AT1,ERK2 expression in rats with chronic heart failure (CHF).Methods The CHF rat models were setup by coronary artery ligation,exhausting swimming and reducing feeding.Divided CHF rat methods into a model group,a lisinopril group,Qi-benefiting with Chinese medicine group,blood activating with Chinese medicine group,Qi benefiting and blood activating with Chinese medicine group(QBBA group) and Guanxin-Shutong capsule group.Rats without left coronary artery ligation were set for sham operated group.Real-time quantitative PCR technique and immunohistochemical method were adopted to detect AT1,ERK2 changes of CHF rats.Results AT1,ERK2 expression increased obviously in the model group compared with the sham operated group,and differences were statistically significant (P＜0.01).After treatment,AT1,ERK2 expression reduced significantly in QBBA group and Guanxin-Shutong capsule group than the lisinopril group.(P＞0.05).And therapeutic effect of Guanxin-Shutong capsule group was much better than other Chinese medicine treated group (P＜0.01).Conclusion Guanxin-Shutong capsule would achieve the goal of treating chronic heart failure By inhibiting the expression of AT1 and ERK2 in organizations,and inhibiting or reversing ventricular remodeling process.

The effects of Guanxinshutong on protection of left ventricular function after acute myocardial infarction in rats / 中华内科杂志

Objective To assess the effects of Guanxinshutong capsule(GXST)on protection of left ventricular(LV)function after acute myocardial infarction(AMI)in rats.Methods Twenty-eight male Sprague Dawley rats were randomized to Model group,Drug group and Sham-operated group,with acute myocardial infarction(AMI)achieved by ligating coronary artery in Model and Drug groups.From one week before surgery to four weeks after surgery,GXST for Drug group(1.5 g/kg,2 times/day)or saline for Model and Sham-operated groups was administered via direct gastric gavage.After four weeks of treatment following surgery,measurement of LV function,pathohistological observation and analysis were performed.Results Compared with rats in the Model group,LV systolic pressure(LVSP)[(97.7 ± 9.0)mm Hg (1 mm Hg =0.133 kPa)vs(85.9 ±9.4)mm Hg],the maximum rising rate of LV pressure(+ dp/dtmax)[(4810.2 ± 595.0)mm Hg/s vs(3786.2 ± 723.0)mm Hg/s]and the maximum dropping rate of LV pressure(-dp/dtmax)[(3781.6 ±573.6)mm Hg/s vs(2774.4 ±633.5)mm Hg/s]in the Drug group were significantly increased,while LV end-diastolic pressure(LVEDP)[(10.3 ± 0.7)mm Hg vs(12.7 ±2.4)mm Hg]in the Drug group was significantly decreased(all P ＜ 0.05).Myocardial pathohistological morphology was improved in the Drug group with fibrosis alleviated[(5.13 ± 1.37)％ vs(7.27 ±1.01)％]and infarct size reduced[(20.14 ± 8.49)％ vs(31.90 ± 4.98)％].Apoptosis index(AI)was decreased[(14.05 ± 4.04)％ vs(20.87 ± 6.03)％]and vessel density was significantly increased by 1.48-fold in the Drug group(all P ＜ 0.05).Conclusions GXST is effective in protecting LV function after AMI in rats,which may be affect through increasing vessel density of infarction area,improving myocardial pathohistological morphology,alleviating fibrosis,reducing infarct size and decreasing AI.

Cardioprotective effects of Guanxinshutong (GXST) against myocardial ischemia/reperfusion injury in rats / 老年心脏病学杂志（英文版）

Background The protective effects against reperfusion injury of cardioprotective drugs have recently been evaluated and found to be inadequate. Guanxinshutong (GXST), a combination of the traditional herb and Mongolian medicine, is effective and safe in treating angina pectoris in clinical trials. We assess the cardioprotective effects of GXST against myocardial ischemia and reperfusion (MI/R) injury in rats and explore its possible mechanism. Methods Forty-five male Sprague Dawley rats were randomized into three groups: non-MI/R group (Sham, n = 15), MI/R group treated with vehicle (Control, n = 15) and MI/R group treated with GXST (Drug, n = 15). MI/R was induced by ligation of the left anterior descending coronary artery (LAD) for 30 minutes, followed by 2/24 hour reperfusion in the Control and Drug groups. In the Sham group, the LAD was exposed without occlusion. GXST powder (in the Drug group) or saline (in the Control and Sham groups) were administered via direct gastric gavage from 7 day prior to surgery. Blood samples were collected from the carotid artery (10 rats each group) after 2 hours of reperfusion, to determine the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1) using enzyme-linked immunosorbent assays. The animals were then sacrificed and the hearts were harvested for histopathology and western blot analysis. Infarct size was measured in the remaining five rats in each group after 24 hours reperfusion. Results GXST significantly decreased levels of TNF-α, IL-1β, IL-6, ICAM-1, apoptosis index (AI) and infarct size. GXST also obviously inhibited nuclear factor kappa B (NF-κB) activity when compared with the Control group (all P < 0.05). Conclusions GXST is effective in protecting the myocardium against MI/R injury in rats. Its possible cardioprotective mechanism involves inhibition of the inflammatory response and apoptosis following MI/R injury.