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.
作者
李彬
王新陆
谢世阳
高原
王幼平
崔琳
王永霞
朱明军
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,Zhengzhou 450000,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第4期1682-1686,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
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)~~
关键词
参附益心方
缺氧
原代心肌细胞
凋亡
Shenfu Yixin Decoction
Anoxic
Primary Cardiomyocytes
Apoptosis
作者简介
通讯作者:朱明军,河南省郑州市金水区人民路19号河南中医药大学第一附属医院,邮编:450000,电话:0371-66289006 E-mail:zhumingjun317@163.com.
