[ERK1/2信号通路对黄芪甲苷IV保护H9c2细胞免受H2O2诱导的氧化损伤的影响]

[Effect of ERK1/2 signaling pathway on astragaloside IV protects H9c2 cells against H2O2-induced oxidative injury].

作者信息

Wang Yuan-yuan, Peng Yang, Zhang Qi, Wu Yan-na, Song Jun-qiu, Liu Yan-xia

机构信息

Department of Pharmacology, Tianjin Medical University, Tianjin 300070, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2011 Aug;27(3):363-7.

PMID:22097737

Abstract

OBJECTIVE

To investigate whether Astragaloside IV(AST) protects H9c2 cells against H2O2-induced oxidative injury partly through ERK1/2 signaling pathway.

METHODS

H9c2 cells oxidative injury was induced by 200 tmol/L H2O2 for 6 hours to establish the H2O2-induced injury model of H9c2 cells. The viability of H9c2 cells was detected using MTf 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 200 micromol/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 AST2O + H2O2 groups (P < 0.01). The activity of LDH in the culture medium was decreased significantly (P < 0.01). The 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.

CONCLUSION

AST protects H9c2 cells against H2O2-induced oxidative injury partly through ERK1/2 signaling pathway.

摘要

目的

探讨黄芪甲苷（AST）是否部分通过细胞外信号调节激酶1/2（ERK1/2）信号通路保护H9c2细胞免受过氧化氢（H2O2）诱导的氧化损伤。

方法

用200 μmol/L H2O2诱导H9c2细胞氧化损伤6小时，建立H9c2细胞H2O2诱导损伤模型。采用噻唑蓝（MTT）法检测H9c2细胞活力。采用比色法检测培养基中乳酸脱氢酶（LDH）活性、总超氧化物歧化酶（T-SOD）、锰超氧化物歧化酶（Mn-SOD）活性及丙二醛（MDA）含量。采用蛋白质免疫印迹法（Western blot）分别检测H9c2细胞中磷酸化ERK1/2（p-ERK1/2）和ERK1/2的表达。

结果

200 μmol/L H2O2处理6小时后，H9c2细胞活力适合后续研究。与H2O2组相比，AST10 + H2O2组和AST20 + H2O2组细胞活力显著升高（P < 0.01）。培养基中LDH活性显著降低（P < 0.01）。T-SOD和Mn-SOD活性显著升高（P < 0.01），MDA含量显著降低（P < 0.01）。用10 mg/L或20 mg/L AST处理后，H2O2损伤的H9c2细胞中p-ERK1/2表达显著增加（P < 0.01），加入ERK1/2抑制剂PD98059后，AST的作用被取消。