参附益心方在缺氧条件下对原代心肌细胞凋亡的干预作用

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英文篇名：Shenfu Yixin Decoction exerts inhibitory effects on the apoptosis of primary cardiomyocytes induced by anoxic condition 作者：李彬 ; 王新陆 ; 谢世阳 ; 高原 ; 王幼平 ; 崔琳 ; 王永霞 ; 朱明军 英文作者：LI Bin;WANG Xin-lu;XIE Shi-yang;GAO Yuan;WANG You-ping;CUI Lin;WANG Yong-xia;ZHU Ming-jun;The First Affiliated Hospital of Henan University of Chinese Medicine; 关键词：参附益心方 ; 缺氧 ; 原代心肌细胞 ; 凋亡 英文关键词：Shenfu Yixin Decoction;;Anoxic;;Primary Cardiomyocytes;;Apoptosis 中文刊名：BXYY 英文刊名：China Journal of Traditional Chinese Medicine and Pharmacy 机构：河南中医药大学第一附属医院; 出版日期：2019-04-01 出版单位：中华中医药杂志 年：2019 期：v.34 基金：国家自然科学基金项目(No.81503419,No.81373853,No.81603466,No.81603432);; 国家重点基础研究发展计划(973计划)(No.2015CB554401);; 河南省创新型科技团队(No.C20130050)~~ 语种：中文; 页：BXYY201904104 页数：5 CN：04 ISSN：11-5334/R 分类号：388-392

摘要

目的:观察参附益心方在缺氧条件下对原代心肌细胞凋亡的干预作用,并探讨其作用机制。方法:取1～3d的SD乳鼠,进行原代心肌细胞分离、培养、鉴定,用缺氧建立心肌细胞凋亡模型,参附益心方0.25、0.5mg/mL、辅酶Q10 1×10-4mol/L干预,CCK-8和Calcein-AM染色检测心肌细胞活性,流式细胞仪检测原代心肌细胞凋亡率,荧光定量PCR检测凋亡相关因子Bcl-2、Bax、Caspase-3、Caspase-8、Caspase-9基因的表达,试剂盒检测凋亡相关因子Caspase-3、Caspase-8、Caspase-9蛋白的活性。结果:与模型组比较,参附益心方高、低剂量组能提高缺氧条件下心肌细胞活性,降低心肌细胞凋亡率(P<0.05,P<0.01);下调促凋亡因子Caspase-3基因及Caspase-3、Caspase-9蛋白的活性,上调抑凋亡因子Bcl-2基因的表达量(P<0.05),参附益心方高剂量组还可下调促凋亡因子Bax、Caspase-9的基因表达(P<0.05),参附益心方对心肌细胞活性、凋亡率、凋亡相关因子的作用呈剂量依赖性。结论:参附益心方可明显干预缺氧条件下原代心肌细胞发生凋亡,其机制可能与调节通过线粒体途径的凋亡相关因子Bcl-2、Bax、Caspase-3、Caspase-9有关。
        Objective: To determine the inhibitory effect of Shenfu Yixin Decoction on anoxic condition induced apoptosis in primary cardiomyocytes and elucidate its underlying mechanism. Methods: After isolated primary cardiomyocytes of SD rats born 1-3 day were cultured and identified, to induce the cardiomyocyte apoptosis model by the anoxic condition. The administration of Shenfu Yixin Decoction(0.25, 0.5 mg/mL) and Coenzyme Q10(1×10-4 mol/L)were then added. Cell activity was detected by CCK-8 and Calcein-AM staining methods, and the cell apoptosis was detected by flow cytometry. The mRNA levels of Bcl-2, Bax, Caspase-3, Caspase-8, Caspase-9 were quantitatively detected by Realtime PCR and the protein levels of Caspase-3, Caspase-8, Caspase-9 were examined by the kit. Results: Compared with the model group, the high and low dose groups of Shenfu Yixin Decoction increased the activity of cardiomyocytes and reduced the apoptosis rate of cardiomyocytes(P<0.05, P<0.01); down-regulated the gene levels of pro-apoptosis factor Caspase-3 and activity of protein Caspase-3, Caspase-9, and up-regulated the gene level of Bcl-2 that was one of key anti-apoptosis factors(P<0.05). The high dose group of Shenfu Yixin Decoction down-regulated the gene levels of pro-apoptosis factor Bax, Caspase-9(P<0.05). The effects of Shenfu Yixin Decoction on myocardial cell activity, apoptosis rate and apoptosis related factors were dose-dependent. Conclusion: Shenfu Yixin Decoction inhibited the apoptosis of primary cardiomyocytes on anoxic condition involving the regulation of apoptosis-related factors including Bcl-2, Bax, Caspase-3, Caspase-9 bymitochondrial pathway.

引文

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