目的 探究人参皂苷Rg₃对H₂O₂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H₂O₂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Rg₃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、qRT-PCR以及Western blotting检测细胞周期、CDK4的mRNA水平和蛋白表达。结果 400 μmol/L H₂O₂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Rg₃浓度的增加,被H₂O₂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。40 μmol/L人参皂苷Rg₃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mRNA水平和蛋白表达水平。结论 人参皂苷Rg₃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, mRNA level and protein express of CDK4 were determined by flow cytometry, qRT-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 mRNA 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.