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Danqi tablet composed of the dried roots of Salvia miltiorrhiza and Panax notoginseng is a well-known Chinese patent medicine commonly used for the treatment of cardio-cerebrovascular diseases such as coronary heart disease and myocardial ischemia. Numerous chemical constituents belonging to S. miltiorrhiza and P. notoginseng were detectable in Danqi tablet. Here, we established and validated a rapid and sensitive ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry method for simultaneous quantification of 23 components in Danqi tablet and then successfully applied to assay 12 batches of samples from ten manufacturers. Our results demonstrated that the contents of 23 components in 12 batches of Danqi tablets varied significantly and their quality indeed existed differently based on the principal component analysis. According to the quantitative data and the loading plot of principal component analysis, five abundant compounds in Danqi tablet were selected as characteristic chemical markers possibly responsible for the quality assessment. Among them, salvianolic acid B and ginsenoside Rg1 were further chosen to be combined at 2:5 ratio to evaluate the anti-thrombotic activity on phenylhydrazine-induced zebrafish heart thrombosis model. Expectedly, this component combination increased the heart red blood cells intensity compared with the model group and the median effective concentration was 123.4 µg/mL, suggestive of its well anti-thrombotic effect. This study contributed to the quantitative evaluation of Danqi tablet and indicated the combination of salvianolic acid B and ginsenoside Rg1 may be capable of reflecting the effect of Danqi tablet, thereby providing a reference for further investigations on the improvement of quality control and clinical application of Danqi tablet.
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Medicamentos Herbarios Chinos/análisis , Animales , Bioensayo , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/uso terapéutico , Espectrometría de Masas , Fenilhidrazinas , Comprimidos/análisis , Trombosis/inducido químicamente , Trombosis/tratamiento farmacológico , Pez CebraRESUMEN
OBJECTIVE: To investigate the cardioprotective effect of Danqi Tablet (DQT, ) on ischemic heart model rats and the regulative effect on energy metabolism through peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α). METHODS: Rat ischemic heart model was induced by ligation of left anterior descending coronary artery. Totally 40 Sprague-Dawley rats were randomly divided into sham group, model group, DQT group (1.5 mg/kg daily) and trimetazidine (TMZ) group (6.3 mg/kg daily) according to a random number table, 10 rats in each group. Twenty-eight days after continuous administration, cardiac function was assessed by echocardiography and the structures of myocardial cells were observed by hematoxylin-eosin staining. The level of adenosine triphosphate (ATP) in myocardial cells was measured by ATP assay kit. Expressions level of key transcriptional regulators, including PGC-1α, Sirtuin 1 (SIRT1), AMP-activated protein kinase (AMPK), and downstream targets of PGC-1α, such as mitofusin 1 (MFN1), mitofusin 2 (MFN2) and superoxide dismutase 2 (SOD2) were measured by Western blot. Expression level of PGC-1α was examined by immunohistochemical staining. RESULTS: The rat ischemic heart model was successfully induced and the heart function in model group was compromised. Compared with the model group, DQT exerted cardioprotective effects, up-regulated the ATP production in myocardial cells and inhibited the infiltration of inflammatory cells in the margin area of infarction of the myocardial tissues (P<0.01). The expressions of PGC-1α, SIRT1 and AMPK were increased in the DQT group (all P<0.05). Furthermore, the downstream targets, including MFN1, MFN2 and SOD2 were up-regulated (P<0.05 or P<0.01). Compared with the TMZ group, the expression levels of PGC-1α, MFN1 and SOD2 were increased by DQT treatment (P<0.05 or P<0.01). CONCLUSION: DQT regulated energy metabolism in rats with ischemic heart model through AMPK/SIRT1 -PGC-1α pathway. PGC-1α might serve as a promising target in the treatment of ischemic heart disease.
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Metabolismo Energético , Proteínas Quinasas Activadas por AMP/metabolismo , Animales , Medicamentos Herbarios Chinos , Miocitos Cardíacos/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Sirtuina 1 , ComprimidosRESUMEN
OBJECTIVE@#To investigate the cardioprotective effect of Danqi Tablet (DQT, ) on ischemic heart model rats and the regulative effect on energy metabolism through peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α).@*METHODS@#Rat ischemic heart model was induced by ligation of left anterior descending coronary artery. Totally 40 Sprague-Dawley rats were randomly divided into sham group, model group, DQT group (1.5 mg/kg daily) and trimetazidine (TMZ) group (6.3 mg/kg daily) according to a random number table, 10 rats in each group. Twenty-eight days after continuous administration, cardiac function was assessed by echocardiography and the structures of myocardial cells were observed by hematoxylin-eosin staining. The level of adenosine triphosphate (ATP) in myocardial cells was measured by ATP assay kit. Expressions level of key transcriptional regulators, including PGC-1α, Sirtuin 1 (SIRT1), AMP-activated protein kinase (AMPK), and downstream targets of PGC-1α, such as mitofusin 1 (MFN1), mitofusin 2 (MFN2) and superoxide dismutase 2 (SOD2) were measured by Western blot. Expression level of PGC-1α was examined by immunohistochemical staining.@*RESULTS@#The rat ischemic heart model was successfully induced and the heart function in model group was compromised. Compared with the model group, DQT exerted cardioprotective effects, up-regulated the ATP production in myocardial cells and inhibited the infiltration of inflammatory cells in the margin area of infarction of the myocardial tissues (P<0.01). The expressions of PGC-1α, SIRT1 and AMPK were increased in the DQT group (all P<0.05). Furthermore, the downstream targets, including MFN1, MFN2 and SOD2 were up-regulated (P<0.05 or P<0.01). Compared with the TMZ group, the expression levels of PGC-1α, MFN1 and SOD2 were increased by DQT treatment (P<0.05 or P<0.01).@*CONCLUSION@#DQT regulated energy metabolism in rats with ischemic heart model through AMPK/SIRT1 -PGC-1α pathway. PGC-1α might serve as a promising target in the treatment of ischemic heart disease.
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We used the network pharmacology to explore the active ingredients and mechanism of action of Danqi tablets in the treatment of coronary heart disease, providing a theoretical basis for the treatment of clinical coronary heart disease. The TCMSP database was used to screen for active ingredients and targets in Danqi tablets; the predicted targets of coronary heart disease were screened through the GeneCards database; then, the intersection of the two targets was mapped; we used STRING database to construct a protein interaction network map and identified 65 cores targets; then, the DAVID database was used for enrichment analysis of gene ontology (GO) biological processes and KEGG signaling pathways, and finally the Cytoscape-3.6.2 software was used to construct a network diagram of traditional Chinese medicine-active ingredient-key targets-pathway. The H2O2-induced H9C2 cells injury model was used for experimental verification. The results suggest that Danqi tablets act on inflammatory factors and apoptosis-related pathways through quercetin, luteolin, tanshinone IIA, and other active ingredients, and improve proinflammatory or anti-inflammatory imbalances in the body and protect cardiomyocytes. This study confirms the multi-component and multi-target effects of Danqi tablets in the treatment of coronary heart disease, and provides an objective basis for their use in further experimental research and the clinical diagnosis and treatment of coronary heart disease.
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[Objective]To establish a HPLC method or the determination of ginsenside Rg1 in Danqi tablet.[Method]HPLC conditions were as fellows:DiamonsiLTM C18 colum,a mobile phase of MeOH:H2O(48:52),and the wavelength at 280nm.[Results] There was a good linearity within the rang of 4.8ug~24.0ug of ginsenside Rg1.The average recover was 99.80%and RSD was 1.46%.[Conclusion]This method can be used for the quality control of Danqi tablet.
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Objective To establish a RP- HPLC method for the determinatio n of protocatechualdehyde in Danqi tablet. Methods LiChrospher RP- 18 column w as used, the mobile phase was MeOH- 1.5 % Hac(1∶ 9) with a flow rate of 1.0 mL/ min, and the detection wavelength was at 280nm. Results There was a goo d linearity within the range of 0.030~ 0.150 ? g of protocatechualdehyde. The average recovery was 103.08 % and RSD was 0.796 % (n=5). Conclusion This method can be used for the quality control of Danqi tablet .
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AIM: To establish a method for the quality control of Danqi Tablet(Radix et Rhizoma Salviae Miltiorrhizae,Radix et Rhizoma Notoginseng,etc). METHODS: The content of salvianic A and protocatechualdehyde,notoginsenoside R_1 and ginsenoside Rg_1 in Danqi Tablet were determined simultaneously respectively by HPLC. RESULTS: The methodological study showed that a good linear correlation existed in the range of 0.164-3.28 ?g(r=0.999 99) of sodium salvianic A,0.013-0.52 ?g(r=0.999 999) of protocatechualdehyde,0.2-6 ?g(r=0.999 997) of notoginsenoside R_1 and 0.824-24.72 ?g(r=0.999 96) of ginsenoside Rg_1,respectively.The average recovery of sodium salvianic A was 98.34%(n=9),RSD was 2.03%,100.65% for protocatechualdehyde(n=9),RSD was 2.96%,98.97% for notoginsenoside R_1(n=9),RSD was 2.81%;98.93% for ginsenoside Rg_1(n=9),RSD was 0.57%. CONCLUSION: The determination methods are simple,accurate and reproducible.The method can be used for quality control of Danqi Tablet.