Rapid identification, quantitation, and antioxidant activity evaluation of the components in Guanxin Shutong capsule with liquid chromatography and mass spectrometry.

Guanxin Shutong capsule (GSC) is a traditional Chinese medicinal prescription used in the treatment of coronary heart disease (CHD) and angina pectoris in clinic. However, the chemical profile of GSC is still uncovered, which hindered the progress of pharmacological study and clinical application. Herein, high performance liquid chromatography (HPLC) together with high resolution mass spectrometry (HR-MS) techniques were employed to analyze the quality consistency and to identify chemical components in GSC. As a result, a total of 111 compounds were tentatively annotated. Quantitative analysis based on HPLC-ultraviolet detection (UV) was performed for 6 main components and fingerprints of 10 different batches of GSC were established. The developed method was validated for linearity, precision, repeatability, stability, and recovery. The quality evaluation and similarity analysis of the 10 batches were also performed. Furthermore, in vitro antioxidant activity assays demonstrated that GSC exhibited potential DPPH and hydroxyl radical scavenging capacities. Especially, salvianolic acids showed the strongest free radical scavenging capacities, which might be the main component for quality control of GSC.

Investigation of the Multi-Target Mechanism of Guanxin-Shutong Capsule in Cerebrovascular Diseases: A Systems Pharmacology and Experimental Assessment.

Guanxin-Shutong capsule (GXSTC), a combination of Mongolian medicines and traditional herbs, has been clinically proven to be effective in treating cerebrovascular diseases (CBVDs). However, the underlying pharmacological mechanisms of GXSTC in CBVDs remain largely unknown. In this study, a combination of systems pharmacology and experimental assessment approach was used to investigate the bioactive components, core targets, and possible mechanisms of GXSTC in the treatment of CBVDs. A total of 15 main components within GXSTC were identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) and a literature research. Fifty-five common genes were obtained by matching 252 potential genes of GXSTC with 462 CBVD-related genes. Seven core components in GXSTC and 12 core genes of GXSTC on CBVDs were further determined using the protein-protein interaction (PPI) and component-target-pathway (C-T-P) network analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis results predicted that the molecular mechanisms of GXSTC on CBVDs were mainly associated with the regulation of the vascular endothelial function, inflammatory response, and neuronal apoptosis. Molecular docking results suggested that almost all of core component-targets have an excellent binding activity (affinity < -5 kcal/mol). More importantly, in middle cerebral artery occlusion (MCAO) -injured rats, GXSTC significantly improved the neurological function, reduced the infarct volume, and decreased the percentage of impaired neurons in a dose-dependent manner. Western blotting results indicated that GXSTC markedly upregulated the expression of vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS), while downregulating the expression of cyclooxygenase-2 (COX-2) and transcription factor AP-1 (c-Jun) in MCAO-injured rats. These findings confirmed our prediction that GXSTC exerts a multi-target synergetic mechanism in CBVDs by maintaining vascular endothelial function, inhibiting neuronal apoptosis and inflammatory processes. The results of this study may provide a theoretical basis for GXSTC research and the clinical application of GXSTC in CBVDs.

The effect of Guanxin Shutong capsule on alleviating the myocardial fibrosis in heart failure rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Guanxin Shutong (GXST) capsule is a renowned traditional Chinese medicine widely used for the treatment of cardiovascular diseases in the clinic. However, no pharmacological experimental studies of GXST has been reported on the treatment of pressure overload-induced heart failure. This study aimed to investigate the effects of GXST capsule on ameliorating myocardial fibrosis conditions in pressure overload-induced heart failure rats. MATERIAL AND METHODS: Rats were randomly divided into 6 groups: Normal group, Model group, GXST-treated group at a dose of 0.5 g/kg, 1 g/kg, 2 g/kg, respectively, and digoxin positive control group at a dose of 1 mg/kg. After 4 weeks of administration, cardiac function was evaluated by echocardiography. Cardiac injury and fibrotic conditions were evaluated by H&E staining, Masson staining, and Sirius Red staining. Myocardial fibrosis was evaluated by immunohistochemistry staining and Western blot. RESULTS: GXST significantly inhibited cardiac fibrosis, reduced the excessive deposition of collagen, and finally improved cardiac function. GXST reversed ventricular remodeling might be through the TGF-ß/Smad3 pathway. CONCLUSION: GXST capsule demonstrated a strong anti-fibrosis effect in heart failure rats by inhibiting the TGF-ß/Smad3 signaling pathway.

Guanxin Shutong Capsule in the Adjuvant Treatment of Unstable Angina Pectoris ：A Meta-analysis and Trial Sequential Analysis / 中国药房

OBJECTIVE： To evaluate clinical efficacy of Guanxin shutong capsule （GSC） in the adjuvant treatment of unstable angina pectoris （UAP）， and to provide evidence-based reference for clinical treatment of UAP. METHODS： Retrieved from PubMed， Embase， Cochrane library， CBM， CNKI， VIP and Wanfang database， randomized controlled trials（RCTs）about routine treatment （trial group） of GSC combined with western medicine versus western medicine routine treatment （control group） in the treatment of UAP were collected during database establishment to Oct. 11th ， 2018. After data extraction of included literatures and quality evaluation with Cochrane bias risk evaluation tool 5.1.0， Meta-analysis of total response rate of angina pectoris， total response rate of ECG， blood lipid levels （TC， HDL-C， LDL-C， TG）， the level of hs-CRP were performed by using Rev Man 5.2 statistical software. TSA 0.9 software was used for trial sequential analysis （TSA） of the total response rate of angina pectoris and response rate of ECG. RESULTS： A total of 11 RCTs were included， involving 946 patients. Results of Meta-analysis showed that total response rate of angina pectoris [RR=1.24，95％CI（1.16，1.32），P＜0.001] and total response rate of ECG [RR＝1.22，95％CI（1.11，1.34），P＜0.001] in trial group were significantly higher than control group. The improvement of TC [SMD＝－1.55，95％CI（－1.81，－1.29），P＜0.001]， TG [SMD＝－0.84，95％CI（－1.08，－0.60），P＜0.001]， HDL-C [SMD＝0.15，95％CI（0.06，0.25），P＝0.001]， LDL-C [SMD＝－0.62，95％CI（－0.76，－0.48），P＜0.001] and hs-CRP [SMD＝－2.54，95％CI（－3.88，－1.88），P＜0.001] in trial group were better than control group. TSA analysis showed that the evidence of Meta-analysis was reliable. CONCLUSIONS： GSC combined with western medicine routine treatment can improve total response rate of angina pectoris， total response rate of ECG， blood lipid and hs-CRP level of UAP patients.

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.

Protective effects of Guanxin Shutong capsule drug-containing serum on tumor necrosis factor-α-induced endothelial dysfunction through nicotinamide adenine dinucleotide phosphate oxidase and the nitric oxide pathway.

The Chinese medicinal formula Guanxin Shutong capsule (GXSTC) has been used for almost 10 years as a clinical treatment for chest pain, depression, palpitation and cardiovascular diseases. The aim of this study was to investigate the effects of GXSTC drug-containing serum on tumor necrosis factor-α (TNF-α)-stimulated endothelial cells. Cell viability was measured by MTT assay, and nitric oxide (NO) levels and NO synthase (NOS) activity were measured as standards of endothelial dysfunction. Malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were evaluated using commercial kits. In addition, the protein expression of endothelial NOS (eNOS), AKT and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits was examined to evaluate the effect of GXSTC drug-containing serum on ECV304 cells. GXSTC significantly reversed the decrease in NO production induced by TNF-α (5 ng/ml) in ECV304 cells. The expression of NADPH oxidase subunits was increased by TNF-α treatment, but markedly inhibited by treatment with GXSTC in TNF-α-stimulated cells. In summary, GXSTC increased the production of NO in ECV304 cells and exerted a protective effect on ECV304 cells stimulated with TNF-α by upregulating the mRNA and protein expression of eNOS. This was accompanied by increased SOD activity and reduced MDA levels. These results suggested that GXSTC protects the endothelium via the NO pathway and exhibits antioxidant effects.

Chinese medicinal formula Guanxin Shutong capsule protects the heart against oxidative stress and apoptosis induced by ischemic myocardial injury in rats.

Guanxin Shutong capsule (GXSTC) is a Chinese medicinal formula that has been used clinically for the treatment of chest pain, depression, palpitation and cardiovascular diseases in China for almost 10 years. The aim of the present study was to investigate the protective mechanisms against oxidative stress and apoptosis that GXSTC exhibits in the hearts of rats with myocardial ischemia (MI). Infarct size and the levels of marker enzymes, including serum creatine kinase-isoenzyme (CK-MB), lactate dehydrogenase (LDH) and glutamate oxaloacetic transaminase (GOT), as well as the levels of nitric oxide (NO) and NO synthase (NOS) in the heart were measured by biochemical analysis assays. Levels of the antioxidants superoxide dismutase (SOD), catalase (CATA), and glutathione (GSH), and the oxidative stress marker malondialdehyde (MDA), were also determined. Following a 6-week period of ischemia, myocardial apoptosis, as well as the protein and mRNA expression of NADPH oxidase, was evaluated. Myocardial NADPH oxidase activity was measured by protein expression of p47phox and gp91phox using western blot analysis and mRNA expression of p22phox, p47phox, p67phox and gp91phox using reverse transcription polymerase chain reaction. The results showed that daily oral treatment of the rats with GXSTC reduced infarct size, myocardial apoptosis, the levels of serum MDA, LDH, CK-MB and GOT and heart GOT, and increased the activities of total SOD, CATA, NOS and the levels of NO and GSH compared with those in vehicle-treated MI model rats. Administration of GXSTC for 6 weeks also reduced the mRNA expression of the NADPH oxidase subunits p47phox and gp91phox protein, as well as the expression of Bax and caspase-3 proteins. By contrast, Bcl-2 protein expression increased. In conclusion, the results demonstrate that GXSTC attenuates myocardial injury via antioxidative and antiapoptotic effects.

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.