人参皂苷Rg_3对H_2O_2诱导人肾小球系膜细胞氧化应激损伤的保护作用及其机制研究

Protection of ginsenoside Rg_3 on oxidative stress injury of human glomerular mesangial cells induced by H_2O_2 and its mechanism

目的探究人参皂苷Rg_3对H_2O_2诱导人肾小球系膜细胞氧化应激损伤的保护作用及其作用机制。方法采用H_2O_2诱导建立人肾小球系膜细胞氧化应激损伤模型。以不同浓度的人参皂苷Rg_3对人肾小球系膜细胞预处理24 h,采用CCK-8试验、乳酸脱氢酶(LDH)试剂盒以及丙二醛(MDA)测定试剂盒分别检测细胞凋亡数量、细胞培养液中LDH水平和MDA含量。通过流式细胞术、q RT-PCR以及Western blotting检测细胞周期、CDK4的m RNA水平和蛋白表达。结果 400μmol/L H_2O_2培养细胞4 h能够建立稳定有效的人肾小球系膜细胞氧化应激损伤模型。随着人参皂苷Rg_3浓度的增加,被H_2O_2损伤的细胞的凋亡数量、细胞外LDH活性、细胞产生的MDA的含量均明显下降。40μmol/L人参皂苷Rg_3可以把人肾小球系膜细胞的细胞周期阻滞在G1期,下调CDK4的m RNA水平和蛋白表达水平。结论人参皂苷Rg_3可以通过抑制CDK4而对人肾小球系膜细胞氧化应激损伤发挥保护作用。

Objective To investigate protective effects of ginsenoside Rg3 on oxidative stress injury of human glomerular mesangial cells induced by H2O2 and to explore its mechanism. Methods Oxidative stress injury models on human glomerular mesangial cells induced by H2O2 were established. Human glomerular mesangial cells were pretreated for 24 h by various concentrations of ginsenoside Rg3, then activity of cells, activity of LDH, and contents of MAD in culture medium were detected by CCK-8 test, LDH kit, and MDA kit. Cell cycle, m RNA level and protein express of CDK4 were determined by flow cytometry, q RT-PCR, and Western blotting. Results Stable and effective models of oxidative stress injury of human glomerular mesangial cells were established by 400 μmol/L H2O2 cultured for 24 h. With the increase of concentrations of ginsenoside Rg3, the amount of apoptotic cells damaged by H2O2 reduced, activity of LDH, and contents of MAD in culture medium were increased. Ginsenoside Rg3 with concentration of 40 μmol/L could block cell cycle in G1 phase, and down-regulate m RNA and protein expresses of CDK4 of human glomerular mesangial cells. Conclusion Ginsenoside Rg3 has protective effect on glomerular mesangial cells against oxidative stress injury of human glomerular mesangial cells induced by H2O2 which may be related to reduction of CDK4.