ERK1/2信号通路对黄芪苷Ⅳ抗H_2O_2诱导H9c2细胞氧化损伤的作用

Effect of ERK1/2 signaling pathway on astragaloside Ⅳ protects H9c2 cells against H_2O_2-induced oxidative injury

目的:研究黄芪苷Ⅳ(AST)是否通过细胞外信号调节激酶1/2(ERK1/2)通路发挥对H2O2诱导的H9c2细胞氧化损伤的保护作用。方法:用200μmol/L的H2O2处理细胞6 h,采用MTT法检测细胞存活率,建立H2O2诱导的H9c2细胞氧化损伤模型;比色法测定细胞培养液中乳酸脱氢酶(LDH)活性、总超氧化物歧化酶(T-SOD)和锰超氧化物歧化酶(Mn-SOD)活力以及丙二醛(MDA)含量;Western blot检测H9c2细胞ERK1/2蛋白的磷酸化水平。结果:在H2O2浓度为200μmol/L作用6 h条件下,细胞存活率降低程度适中,实验结果重复性好,确定后续实验采用200μmol/L H2O2作用6 h建立模型。与H2O2组比较,10 mg/L及20 mg/L AST均显著提高细胞存活率(P<0.01),使细胞培养液中LDH活性显著降低(P<0.01),T-SOD及Mn-SOD活力显著提高(P<0.01),MDA含量显著降低(P<0.01)。10 mg/L及20 mg/L AST均显著增加H2O2损伤的H9c2细胞p-ERK1/2蛋白的表达(P<0.01),当用PD98059(ERK1/2的抑制剂)预处理后,AST的作用则被取消。结论:黄芪苷Ⅳ可以通过ERK1/2通路发挥对H2O2诱导的H9c2细胞氧化损伤的保护作用。

Objective： To investigate whether Astragaloside Ⅳ（AST） protects H9c2 cells against H2O2-induced oxidative injury partly through ERK1/2 signahng pathway. Methods： H9c2 cells oxidative injury was induced by 200 μmol/L H2O2 for 6 hours to establish the H2O2- induced injury model of H9c2 cells. The viability of H9c2 cells was detected using MTT method. Activity of lactate dehydrogenase（LDH）, total-superoxide dismutase（T-SOD）, manganese-superoxide dismutase（Mn-SOD） and content of MDA （ malondialdehyde ） in the culture medium were detected using colorimetric method. Western blot was performed to exam expression of p-ERK1/2 and ERK1/2 in H9c2 cells respectively. Results： Under 200tanol/L H2O2 treatment for 6 hours, the vaibility of H9c2 cells was suitable for the following study. Compared with H2O2 group, the cell viability was increased significantly in AST10 ＋ H2O2 and AST20 ＋ H2O2 groups（ P 〈 0.01）. The activity of LDH in the culture medium was decreased significantly（ P 〈 0.01）. 11ae activity of T-SOD and Mn-SOD was increased significantly（ P 〈 0.01）, the content of MDA was decreased significantly（ P 〈0.01）. Treated with 10 mg/L or 20 mg/L of AST, expression of p-ERK1/2 in H9c2 cells injured from H2O2 was increased significantly（ P 〈 0.01）, when PD98059（inhibitor of ERK1/2） was added, the effects of AST were cancelled. Condusion： AST protects H9c2 cells against H2O2-induced oxidative injury partly through ERK1/2 signaling pathway.