黄芪注射液对血管紧张素Ⅱ诱导的H9c2心肌细胞凋亡的影响及其机制研究

Effect of Astragalus Injection on AngiotensinⅡ-Induced H9c2 Myocardial Apoptosis and Its Mechanism

【目的】探讨黄芪注射液对血管紧张素Ⅱ(AngⅡ)诱导的H9c2心肌细胞凋亡的影响及可能机制。【方法】将H9c2心肌细胞分为对照组、AngⅡ组、AngⅡ+黄芪注射液组。以AngⅡ诱导的心肌细胞凋亡为模型。细胞计数试剂盒8(CCK-8)检测细胞活性;流式细胞术测定细胞凋亡情况;三磷酸腺苷(ATP)比色/荧光法检测细胞中ATP含量的变化;钙离子荧光探针(Fura-2AM)法检测细胞内钙离子含量;实时定量聚合酶链反应(RT-PCR)法检测细胞电压依赖性阴离子通道1(VDAC1)、线粒体融合蛋白2(Mfn2)及葡萄糖调节蛋白75(GRP75)基因的表达;蛋白免疫印迹(Western Blot)法检测细胞VDAC1、Mfn2、GRP75的蛋白表达。【结果】与对照组比较,AngⅡ组细胞凋亡率,ATP含量,钙含量,Mfn2、GRP75 mRNA表达水平,VDAC1、Mfn2、GRP75蛋白表达水平升高,VDAC1 m RNA表达水平降低(P<0.05或P<0.01);与AngⅡ组比较,AngⅡ+黄芪注射液组细胞凋亡率,ATP含量,钙含量,Mfn2、GRP75 m RNA表达水平,VDAC1、Mfn2、GRP75蛋白表达水平降低,VDAC1 mRNA表达水平升高(P<0.05或P<0.01)。【结论】黄芪注射液可通过改善心肌细胞能量代谢和线粒体钙离子转运来抑制心肌细胞凋亡,发挥心肌细胞保护作用。

Objective To investigate the effect of Astragalus injection on the angiotensinⅡ-induced H9c2myocardial apoptosis and its possible mechanism.Methods H9c2 cardiomyocytes were divided into control group,AngⅡgroup and AngⅡ+Astragalus injection group.Myocardial apoptosis induced by AngⅡwas used as a model.Cell activity was determined by cell counting kit-8(CCK-8);cell apoptosis was determined by flow cytometry(FCM);adenosine triphosphate(ATP)content was detected by colorimetric/fluorescence assay;intracellular calcium ion content was detected by calcium fluorescence probe(Fura-2AM).The expression of voltage-dependent anionic channel 1(VDAC1),mitochondrial fusion protein 2(Mfn2)and glucose regulatory protein 75(GRP75)genes were detected by real-time quantitative polymerase chain reaction(RT-PCR).The protein expression of VDAC1,Mfn2 and GRP75 were detected by Western Blot.Results Compared with the control group,the apoptosis rate,ATP content,calcium content,Mfn2 and GRP75 mRNA expression levels,and VDAC1,Mfn2and GRP75 protein expression levels were increased and VDAC1 mRNA level was decreased in the AngⅡgroup(P<0.05 or P<0.01).Compared with the AngⅡgroup,the apoptosis rate,ATP content,calcium content,Mfn2 and GRP75 mRNA expression levels,and VDAC1,Mfn2 and GRP75 protein expression levels were decreased,and VDAC1 mRNA expression level was increased in AngⅡ+Astragalus injection group(P<0.05 or P<0.01).Conclusion Astragalus injection is able to inhibit the myocardial apoptosis by improving the energy metabolism and mitochondrial calcium ion transport.