Objective: To assess the effects of traditional Chinese medicine YiQiFuMai on cardiac function during the progression of ischemic heart failure. Methods: Rabbits were divided into sham, heart failure, and YiQiFuMai gr...Objective: To assess the effects of traditional Chinese medicine YiQiFuMai on cardiac function during the progression of ischemic heart failure. Methods: Rabbits were divided into sham, heart failure, and YiQiFuMai groups. The ischemic heart failure model was established in New Zealand white rabbits, which were intraperitoneally injected with YiQiFuMai injection and 0.9% sodium chloride after the operation. After six weeks, cardiac function was examined by ultrasound;serum BNP levels were measured by ELISA;p-AKT, eNOS, ICAM-1 and VEGF levels were evaluated by real-time PCR and Western-Blot;pathological changes of the myocardial tissue were observed by H&E staining;CD31 expression in tissue samples was analyzed by immunohistochemistry. The ultrastructure and microcirculation of myocardial tissue specimens from the three groups were assessed by transmission electron microscopy. Results: YiQiFuMai decreased serum BNP levels, and increased LVEF and reduced LVEDD at 6 weeks postoperatively. In addition, YiQiFuMai can improve myocardial damage and microcirculation structure, as assessed by histology and transmission electron microscope. At the molecular level, treatment with YiQiFuMai resulted in increased eNOS, VEGF and p-AKT levels but reduced ICAM-1 amounts compared with the heart failure group. Conclusion: Ischemic heart failure damages the microvascular structure and functions of the myocardium. Treatment with YiQiFuMai potentially ameliorates microcirculatory damage and alleviates cardiac failure by improving endothelial function and angiogenesis, and inhibiting inflammatory cell adhesion.展开更多
The YiQiFuMai powder injection （ YQFM）, a Traditional Chinese Medicine （TCM） prescription re-de- veloped based on the well-known TCM formula Sheng-maisan, showed a wide range of pharmacological activities in ca...The YiQiFuMai powder injection （ YQFM）, a Traditional Chinese Medicine （TCM） prescription re-de- veloped based on the well-known TCM formula Sheng-maisan, showed a wide range of pharmacological activities in cardiovascular diseases in clinic. However, its role in protection against myocardial ischemia/reperfusion （MI/R） injury has not been elucidated. The present study not only evaluated the eardioprotective effect of YQFM from MI/ R injury but also investigated the potential molecular mechanisms in vivo and in vitro. The mouse model of MI/R injury was induced by a transient vessel occlusion for 30 rain and reperfusion for 24 h. The myocardium infarct size, production of lactate dehydrogenase （LDH）, creatine kinase （CK）, TUNEL staining and easpase-3 activity were measured. AMPKeα and phospho-AMPKα was analyzed by Western blot. We further verified the protective effect and potential molecular mechanisms of YQFM in an in vitro model of simulated ischemia and reperfusion （ SI/ R） in H9c2 cardiomyocytes. Cell viability was determined, and cell apoptosis were measured by Hoechst 33342 staining and Flow cytometry. Mitochondrial membrane potential （△ψFm） was measured, and ATP content was quantified by biolumineseent assay. Expression of apoptosis-related proteins including Caspase-3, Bcl-2, Bax, AMPKα and phospho-AMPKα was analyzed by Western blot. AMPKoL siRNA transfection was also applied to the mechanism elucidation. YQFM significantly reduced myocardium infarct size and the production of LDH, CK in se- rum, and also produced a significant decrease of apoptotic index which was confirmed by TUNEL staining and the changes of caspase-3 activity. In addition, pretreatment with YQFM markedly improved cell viability and decreased LDH release. Moreover, YQFM inhibited H9c2 apoptosis, blocked the expression of easpase-3 and modulated Bcl- 2 and Bax proteins, leading to an increased mitochondrial membrane potential and cellular ATP content. Mechanis- tically, YQFM activated AMPK signaling pathways while pretreatment with AMPK inhibitor compound C and appli- cation of transfection with AMPKα siRNA attenuated the anti-apoptotie effect of YQFM. Our results indicated that YQFM could provide significant cardioproteetion against MI/R injury, and potential mechanisms might to suppres- sion of cardiomyocytes apoptosis at least in part through activating the AMPK signaling pathways.展开更多
Aim YiQiFuMai Powder Injection is a well-known traditional Chinese medicine formula that has been used extensively in clinical treatment of cardio-cerebral ischemic diseases in China. However, the mechanisms under-lyi...Aim YiQiFuMai Powder Injection is a well-known traditional Chinese medicine formula that has been used extensively in clinical treatment of cardio-cerebral ischemic diseases in China. However, the mechanisms under-lying its clinical efficacy remain unknown. In this study, a network pharmacology approach was employed to identify the YiQiFuMai Powder Injection＇s potential pathways and targets against cardio-cerebral ischemia. The target-path- way interaction network clustered the signaling pathways based on high degree nodes of the drug-target network. The potential protein targets presented in the highly scored clustered pathways were the key network hubs and concentrated on one or limited functional signaling pathways amenable to experimental verification. Twelve main functional annota- tion clusters and main signaling pathways for YiQiFuMai Powder Injection were established by Biocarta analysis, in- eluding the NF-KB signaling pathway, the MAPKinase signaling pathway and the mTOR-signaling pathway and so on. YiQiFuMai Powder Injection is hypothesized to target multiple proteins with a high degree and betweenness of net- work. In addition, the most related pathways were also confirmed in tumor necrosis factor-alpha （TNF-oL） induced human vascular endothelial cell line EA. hy926 by Western blot. This study elucidates the systematic network and pathway analysis of multi-targets in YiQiFuMai Powder Injection. The results provide the possible mechanisms for its mode of action against cardio-cerebral ischemic diseases and may also reveal new clues for its potential application in the inflammatory diseases or tumors.展开更多
Objective To predict the main bioactive components and potential mechanisms of Yiqifumai formula on heart failure by network pharmacology.Methods The bioactive compounds of Yiqifumai formula were screened by TCMSP dat...Objective To predict the main bioactive components and potential mechanisms of Yiqifumai formula on heart failure by network pharmacology.Methods The bioactive compounds of Yiqifumai formula were screened by TCMSP database.The target genes of heart failure were obtained by entering PharmMapper and GeneCards database.R 3.6.3 was used to intercept intersection network for candidate targets.The network of“drug-compound-target-disease”was established via Cytoscape 3.7.2 and STRING platform.The DAVID database was used for GO enrichment analysis and KEGG pathway based on the candidate targets.Results Eleven key compounds(such as Schizandrin C,Gomisin G,ginsenoside rh2,Ruscogenin etc.,),426 non-repeated targets and 382 heart failure targets were obtained from Yiqifumai formula.There were 565 GO items(P<0.05),among which 412 were biological process(BP)items,63 were cell components(CC)items,and 93 were molecular function(MF)items.One hundred and seventeen signal pathways were enriched by KEGG pathway(P<0.05).Conclusion The bioactive compounds in Yiqifumai formula can play the“multi-target”role of enhancing resistance and eliminating pathogenic factors via regulation of angiogenesis and immune in the treatment of heart failure.展开更多
目的:建立注射用益气复脉(冻干)总木脂素的含量测定方法。方法:采用Phenomenex luna C18色谱柱(250mm×4.6mm,5μm);流动相:乙腈-水梯度洗脱;柱温30℃;检测波长218nm;进样量10μL;流速1mL/min。结果:五味子醇甲进样量在0.096～0.578...目的:建立注射用益气复脉(冻干)总木脂素的含量测定方法。方法:采用Phenomenex luna C18色谱柱(250mm×4.6mm,5μm);流动相:乙腈-水梯度洗脱;柱温30℃;检测波长218nm;进样量10μL;流速1mL/min。结果:五味子醇甲进样量在0.096～0.578μg范围内线性关系良好,其回归方程为:Y=404589x-9426.4,r=0.9998,精密度RSD1.88%,稳定性RSD2.29%,重复性试验RSD1.49%,平均回收率为99.84%、RSD 2.08%。结论:实验结果表明该方法简便,灵敏,专属性强,重复性好,可作为注射用益气复脉(冻干)的质量控制方法。展开更多
目的建立同时测定益气复脉方中芍药苷、甘草苷和甘草酸含量的方法。方法采用HPLC法测定芍药苷、甘草苷和甘草酸的含量,Purospher RP C_(18)色谱柱(250mm×4.6mm,5μm);乙腈-1mL·L^(-1)磷酸进行梯度洗脱;流速为1.0mL·min^(...目的建立同时测定益气复脉方中芍药苷、甘草苷和甘草酸含量的方法。方法采用HPLC法测定芍药苷、甘草苷和甘草酸的含量,Purospher RP C_(18)色谱柱(250mm×4.6mm,5μm);乙腈-1mL·L^(-1)磷酸进行梯度洗脱;流速为1.0mL·min^(-1);检测波长为230和237nm;柱温为30℃;进样量为10μL。结果芍药苷、甘草苷和甘草酸的质量浓度分别在0.02~0.31,0.02~0.32和0.02~0.30mg·mL^(-1)范围内线性关系良好,r值均为0.999 9,3种成分的平均回收率分别为97.98%,98.23%和97.49%,RSD值均小于3.00%。结论该方法简便、准确、重复性好,可用于益气复脉方的内在质量控制。展开更多
文摘Objective: To assess the effects of traditional Chinese medicine YiQiFuMai on cardiac function during the progression of ischemic heart failure. Methods: Rabbits were divided into sham, heart failure, and YiQiFuMai groups. The ischemic heart failure model was established in New Zealand white rabbits, which were intraperitoneally injected with YiQiFuMai injection and 0.9% sodium chloride after the operation. After six weeks, cardiac function was examined by ultrasound;serum BNP levels were measured by ELISA;p-AKT, eNOS, ICAM-1 and VEGF levels were evaluated by real-time PCR and Western-Blot;pathological changes of the myocardial tissue were observed by H&E staining;CD31 expression in tissue samples was analyzed by immunohistochemistry. The ultrastructure and microcirculation of myocardial tissue specimens from the three groups were assessed by transmission electron microscopy. Results: YiQiFuMai decreased serum BNP levels, and increased LVEF and reduced LVEDD at 6 weeks postoperatively. In addition, YiQiFuMai can improve myocardial damage and microcirculation structure, as assessed by histology and transmission electron microscope. At the molecular level, treatment with YiQiFuMai resulted in increased eNOS, VEGF and p-AKT levels but reduced ICAM-1 amounts compared with the heart failure group. Conclusion: Ischemic heart failure damages the microvascular structure and functions of the myocardium. Treatment with YiQiFuMai potentially ameliorates microcirculatory damage and alleviates cardiac failure by improving endothelial function and angiogenesis, and inhibiting inflammatory cell adhesion.
文摘The YiQiFuMai powder injection （ YQFM）, a Traditional Chinese Medicine （TCM） prescription re-de- veloped based on the well-known TCM formula Sheng-maisan, showed a wide range of pharmacological activities in cardiovascular diseases in clinic. However, its role in protection against myocardial ischemia/reperfusion （MI/R） injury has not been elucidated. The present study not only evaluated the eardioprotective effect of YQFM from MI/ R injury but also investigated the potential molecular mechanisms in vivo and in vitro. The mouse model of MI/R injury was induced by a transient vessel occlusion for 30 rain and reperfusion for 24 h. The myocardium infarct size, production of lactate dehydrogenase （LDH）, creatine kinase （CK）, TUNEL staining and easpase-3 activity were measured. AMPKeα and phospho-AMPKα was analyzed by Western blot. We further verified the protective effect and potential molecular mechanisms of YQFM in an in vitro model of simulated ischemia and reperfusion （ SI/ R） in H9c2 cardiomyocytes. Cell viability was determined, and cell apoptosis were measured by Hoechst 33342 staining and Flow cytometry. Mitochondrial membrane potential （△ψFm） was measured, and ATP content was quantified by biolumineseent assay. Expression of apoptosis-related proteins including Caspase-3, Bcl-2, Bax, AMPKα and phospho-AMPKα was analyzed by Western blot. AMPKoL siRNA transfection was also applied to the mechanism elucidation. YQFM significantly reduced myocardium infarct size and the production of LDH, CK in se- rum, and also produced a significant decrease of apoptotic index which was confirmed by TUNEL staining and the changes of caspase-3 activity. In addition, pretreatment with YQFM markedly improved cell viability and decreased LDH release. Moreover, YQFM inhibited H9c2 apoptosis, blocked the expression of easpase-3 and modulated Bcl- 2 and Bax proteins, leading to an increased mitochondrial membrane potential and cellular ATP content. Mechanis- tically, YQFM activated AMPK signaling pathways while pretreatment with AMPK inhibitor compound C and appli- cation of transfection with AMPKα siRNA attenuated the anti-apoptotie effect of YQFM. Our results indicated that YQFM could provide significant cardioproteetion against MI/R injury, and potential mechanisms might to suppres- sion of cardiomyocytes apoptosis at least in part through activating the AMPK signaling pathways.
文摘Aim YiQiFuMai Powder Injection is a well-known traditional Chinese medicine formula that has been used extensively in clinical treatment of cardio-cerebral ischemic diseases in China. However, the mechanisms under-lying its clinical efficacy remain unknown. In this study, a network pharmacology approach was employed to identify the YiQiFuMai Powder Injection＇s potential pathways and targets against cardio-cerebral ischemia. The target-path- way interaction network clustered the signaling pathways based on high degree nodes of the drug-target network. The potential protein targets presented in the highly scored clustered pathways were the key network hubs and concentrated on one or limited functional signaling pathways amenable to experimental verification. Twelve main functional annota- tion clusters and main signaling pathways for YiQiFuMai Powder Injection were established by Biocarta analysis, in- eluding the NF-KB signaling pathway, the MAPKinase signaling pathway and the mTOR-signaling pathway and so on. YiQiFuMai Powder Injection is hypothesized to target multiple proteins with a high degree and betweenness of net- work. In addition, the most related pathways were also confirmed in tumor necrosis factor-alpha （TNF-oL） induced human vascular endothelial cell line EA. hy926 by Western blot. This study elucidates the systematic network and pathway analysis of multi-targets in YiQiFuMai Powder Injection. The results provide the possible mechanisms for its mode of action against cardio-cerebral ischemic diseases and may also reveal new clues for its potential application in the inflammatory diseases or tumors.
文摘Objective To predict the main bioactive components and potential mechanisms of Yiqifumai formula on heart failure by network pharmacology.Methods The bioactive compounds of Yiqifumai formula were screened by TCMSP database.The target genes of heart failure were obtained by entering PharmMapper and GeneCards database.R 3.6.3 was used to intercept intersection network for candidate targets.The network of“drug-compound-target-disease”was established via Cytoscape 3.7.2 and STRING platform.The DAVID database was used for GO enrichment analysis and KEGG pathway based on the candidate targets.Results Eleven key compounds(such as Schizandrin C,Gomisin G,ginsenoside rh2,Ruscogenin etc.,),426 non-repeated targets and 382 heart failure targets were obtained from Yiqifumai formula.There were 565 GO items(P<0.05),among which 412 were biological process(BP)items,63 were cell components(CC)items,and 93 were molecular function(MF)items.One hundred and seventeen signal pathways were enriched by KEGG pathway(P<0.05).Conclusion The bioactive compounds in Yiqifumai formula can play the“multi-target”role of enhancing resistance and eliminating pathogenic factors via regulation of angiogenesis and immune in the treatment of heart failure.
文摘目的:建立注射用益气复脉(冻干)总木脂素的含量测定方法。方法:采用Phenomenex luna C18色谱柱(250mm×4.6mm,5μm);流动相:乙腈-水梯度洗脱;柱温30℃;检测波长218nm;进样量10μL;流速1mL/min。结果:五味子醇甲进样量在0.096～0.578μg范围内线性关系良好,其回归方程为:Y=404589x-9426.4,r=0.9998,精密度RSD1.88%,稳定性RSD2.29%,重复性试验RSD1.49%,平均回收率为99.84%、RSD 2.08%。结论:实验结果表明该方法简便,灵敏,专属性强,重复性好,可作为注射用益气复脉(冻干)的质量控制方法。
文摘目的建立同时测定益气复脉方中芍药苷、甘草苷和甘草酸含量的方法。方法采用HPLC法测定芍药苷、甘草苷和甘草酸的含量,Purospher RP C_(18)色谱柱(250mm×4.6mm,5μm);乙腈-1mL·L^(-1)磷酸进行梯度洗脱;流速为1.0mL·min^(-1);检测波长为230和237nm;柱温为30℃;进样量为10μL。结果芍药苷、甘草苷和甘草酸的质量浓度分别在0.02~0.31,0.02~0.32和0.02~0.30mg·mL^(-1)范围内线性关系良好,r值均为0.999 9,3种成分的平均回收率分别为97.98%,98.23%和97.49%,RSD值均小于3.00%。结论该方法简便、准确、重复性好,可用于益气复脉方的内在质量控制。
