黄芪甲苷对肾小球系膜细胞氧化应激损伤的保护作用及其机制  

Protective effect and mechanism of astragaloside Ⅳ on oxidative stress injury of mesangial cells

作　　者：曹玲玲[1,2] 李维祖[1] 司秀莲[1] 孙立[1] 李卫平[1,3] 

机构地区：[1]安徽医科大学药理教研室,安徽合肥230032 [2]淮南联合大学,安徽淮南232038 [3]安庆医药高等专科学校,安徽安庆246052

出　　处：《中国中药杂志》2013年第5期725-730,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81173624);安徽省科技厅国际合作项目(12030603007);安徽省自然科学基金项目(11040606M201);安徽医科大学博士启动基金(XJ201011);安徽省教育厅重点项目(KJ2012A192)

摘　　要：目的:研究黄芪甲苷(astragalosideⅣ,ASⅣ)对H2O2诱导肾小球系膜细胞(human mesangial cells,HMC)氧化应激损伤的保护作用,并进一步探讨其分子机制。方法:将培养的肾小球系膜细胞随机分为5组:正常对照组、H2O2模型组、ASⅣ(12.5,100 nmol.L-1)组、阳性药(Tempol,1×105nmol.L-1)组。采用MTT法观察细胞活力;Hoechst 33258染色检测细胞凋亡;DHE染色检测胞内活性氧(reactive oxygen species,ROS)的产生;流式细胞术检测细胞周期变化情况;Western blot方法测定细胞周期蛋白Cyclin D1,Cyclin A及磷酸化p38,T-p38的表达。结果:1×105,2×105,3×105,4×105nmol.L-1H2O2均能诱导肾小球系膜细胞发生氧化应激损伤:细胞存活率明显降低,细胞活力与H2O2浓度及其作用的时间呈正相关下降趋势。100 nmol.L-1黄芪甲苷能够显著减轻H2O2(3×105nmol.L-1)诱导的肾小球系膜细胞氧化应激损伤:提高细胞存活率及细胞活力,抑制细胞凋亡,降低胞内ROS产量;恢复细胞周期蛋白Cyclin D1表达及各细胞周期的百分比,恢复细胞的正常增殖;降低磷酸化p38的表达。结论:黄芪甲苷对H2O2诱导肾小球系膜细胞氧化应激损伤有一定的保护作用。其机制可能与抑制p38/MAPK信号通路,影响细胞周期蛋白Cyclin D1的表达及降低H2O2诱导的细胞内ROS氧化应激损伤有关。Objective： To study the protective effect of astragaloside Ⅳ （AS Ⅳ ） on H2 02 induced human mesangial cells （HMC）, and further explore its molecular mechanism. Method： The cultured mesangial cells were divided into 5 groups： the normal control group, the H202model group, the AS IV （ 12. 5, 100 nmol ·L-1 ） group and the Tempol （ 1 × 105 nmol· L-1 ） group. The MTr method was used to observe cell viability. Hoechst 33258 staining was used to observe the HMC apoptosis and DHE staining was used to detect the generation of reactive oxygen species （ROS）. The flow cytometry was used to detect the changes in cell cycle. West- em blot was used to detect the expression of Cychn D1, CycllnA, p38, and T-p38. Result： H2O2（1 × 105 , 2 × 105 , 3 × 105 , and 4 × 105 nmol ·L^-1 ） could induce HMC oxidative stress injury, with significant decrease in the cell survival rate. AS IV （100 nmol ·L^-1 ） could significantly inhibit HMC oxidative stress injury induced by H2 02 （3 × 105 nmol ·L^-1 ）, increase the survival rate of HMC cells, inhibit cell apoptosis, and decrease intracellular ROS production. AS IV could also increase the expression of Cyclin D1, recover normal cell proliferation, and decrease the expression of p38. Conclusion： AS IV can protect H2 02induced oxidative stress injury in mesangial cells. Its mechanisms may be related to inhibiting the p38/MAPK signaling pathway, increasing the expression of Cyclin D1 and decreasing the intracellular ROS oxidative stress injury.

关 键 词：黄芪甲苷 过氧化氢 氧化应激 ROS CYCLIN D1 肾小球系膜细胞 

分 类 号：R285.5[医药卫生—中药学]