目的 探究人參皂苷Rg₃對(duì)H₂O₂誘導(dǎo)人腎小球系膜細(xì)胞氧化應(yīng)激損傷的保護(hù)作用及其作用機(jī)制。方法 采用H₂O₂誘導(dǎo)建立人腎小球系膜細(xì)胞氧化應(yīng)激損傷模型。以不同濃度的人參皂苷Rg₃對(duì)人腎小球系膜細(xì)胞預(yù)處理24 h,采用CCK-8試驗(yàn)、乳酸脫氫酶(LDH)試劑盒以及丙二醛(MDA)測(cè)定試劑盒分別檢測(cè)細(xì)胞凋亡數(shù)量、細(xì)胞培養(yǎng)液中LDH水平和MDA含量。通過流式細(xì)胞術(shù)、qRT-PCR以及Western blotting檢測(cè)細(xì)胞周期、CDK4的mRNA水平和蛋白表達(dá)。結(jié)果 400 μmol/L H₂O₂培養(yǎng)細(xì)胞4 h能夠建立穩(wěn)定有效的人腎小球系膜細(xì)胞氧化應(yīng)激損傷模型。隨著人參皂苷Rg₃濃度的增加,被H₂O₂損傷的細(xì)胞的凋亡數(shù)量、細(xì)胞外LDH活性、細(xì)胞產(chǎn)生的MDA的含量均明顯下降。40 μmol/L人參皂苷Rg₃可以把人腎小球系膜細(xì)胞的細(xì)胞周期阻滯在G1期,下調(diào)CDK4的mRNA水平和蛋白表達(dá)水平。結(jié)論 人參皂苷Rg₃可以通過抑制CDK4而對(duì)人腎小球系膜細(xì)胞氧化應(yīng)激損傷發(fā)揮保護(hù)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.