氧化苦参碱对高糖引起的H9C2细胞氧化应激损伤的影响

Effect of Oxymatrine on Oxidative Stress Injury of H9C2 Cells Induced by High Glucose

目的:研究氧化苦参碱对高糖诱导H9C2细胞氧化应激损伤的保护机制。方法:分组培养H9C2心肌细胞并分为空白组,高糖组,氧化苦参碱低、高剂量组(50,100 mg·L^-1),阳性药维生素E组(1×10^-4 mol·L^-1),甘露醇等渗组。通过乳酸脱氢酶外漏法检测细胞损伤,检测细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量变化,通过荧光探针检测细胞内活性氧(ROS)含量和细胞线粒体功能的完整性,蛋白免疫印迹法(Western blot)检测B细胞淋巴瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax)蛋白的表达。结果:与空白组比较,模型组细胞内MDA,ROS含量,Bax蛋白表达水平显著升高(P<0.01),而SOD活性、线粒体膜电位和Bcl-2蛋白表达水平显著降低(P<0.01)。与高糖组比较,氧化苦参碱显著降低乳酸脱氢酶的外泄、显著抑制细胞内ROS的产生(P<0.01),明显降低MDA的含量和Bax蛋白的表达(P<0.05),增加SOD活性、线粒体膜电位和Bcl-2蛋白的表达(P<0.01)。结论:氧化苦参碱通过改善线粒体功能来调节氧化应激从而抑制由高糖引起的H9C2心肌细胞凋亡。

Objective:To study the protective mechanism of oxymatrine on oxidative stress induced by high glucose in H9C2 cells.Method:H9C2 cardiomyocytes were cultured in groups and divided into normal group,high glucose(HG)group,low-dose oxymatrine(OMT)group(50 mg·L^-1),high-dose OMT group(100 mg·L^-1),positive drug vitamin E(VE)group(1×10^-4 mol·L^-1)and mannitol(M)wasotonic control group.Cell damage was detected by lactate dehydrogenase leakage,changes in cell superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected,intracellular reactive oxygen species(ROS)content and cellular mitochondria and functional integrity were detected by fluorescent probes,and Western blotting was used to detect the expressions of Bcl family proteins.Result:Compared with the normal group,the content of malondialdehyde and reactive oxygen species and the expression level of pro-apoptotic protein in the high glucose group were significantly increased(P<0.01),while the activity of superoxide dismutase and the expression levels of mitochondrial membrane potential(MMP)and anti-apoptotic protein were significantly decreased(P<0.01).Compared with the high glucose group,oxymatrine significantly reduced the leakage of lactate dehydrogenase,significantly inhibited the production of intracellular ROS(P<0.01),reduced the amount of malondialdehyde and down-regulated the expression of pro-apoptotic protein(P<0.05),increased the activity of superoxide dismutase,regulated MMP and improved the expression of antiapoptotic protein(P<0.01).Conclusion:Oxymatrine can regulate oxidative stress by improving mitochondrial function,so as to inhibit the apoptosis of H9C2 cardiomyocytes induced by high glucose.