目的 探讨人参皂苷Rg3 对过氧化氢诱导的人晶状体上皮细胞氧化损伤的改善作用及对核转录因子E2 相关因子 2(nuclear factor E2 related factor 2，Nrf2)/血红素加氧酶-1(heme oxygenase 1，HO-1)信号通路的调节作用。方法 用不同浓度人参皂苷Rg3 处理过氧化氢诱导的SRA01/04细胞，用噻唑盐(methyl thiazolyl tetrazolium，MTT)法检测细胞存活率。将第3代对数生长期SRA01/04细胞随机分为正常组、氧化损伤组(用200 μmol∙mL-1过氧化氢处理)、人参皂苷Rg3低剂量组和人参皂苷Rg3高剂量组(分别用40、80 μg∙mL-1人参皂苷Rg3处理6 h，更换培养基后用200 μmol∙mL-1 过氧化氢处理12 h)，用MTT法检测细胞存活率，用流式细胞仪检测细胞凋亡率，用试剂盒检测丙二醛(malondialdehyde，MDA)、超氧化物歧化酶(superoxidedismutase，SOD)和谷胱甘肽过氧化物酶(glutathioneperoxidase，GSH-Px)的含量，用蛋白印迹法检测Nrf2、Kelch样环氧氯丙烷相关蛋白1(Kelch like epichlorohydrin related protein 1，Keap1)和HO-1蛋白的相对表达量。结果 与 0 μg∙mL-1 人参皂苷Rg3 组比较，10、20、40、80 μg∙mL-1 人参皂苷Rg3组的细胞存活率逐渐升高(P＜0。05)。与正常组比较，氧化损伤组的细胞存活率、SOD和GSH-Px含量以及Nrf2、Keap1和HO-1蛋白相对表达量降低，细胞凋亡率和MDA含量升高(P＜0。05);与氧化损伤组比较，人参皂苷Rg3低剂量和人参皂苷Rg3高剂量组的细胞存活率、SOD和GSH-Px含量以及Nrf2、Keap1和HO-1蛋白的相对表达量升高，细胞凋亡率和MDA含量降低(P＜0。05);人参皂苷Rg3低剂量和人参皂苷Rg3高剂量组各项指标水平变化规律相同，人参皂苷Rg3高剂量组更显著(P＜0。05)。结论 人参皂苷Rg3可抑制过氧化氢诱导的人晶状体上皮细胞凋亡，减轻氧化应激损伤，其可能是通过激活Nrf2/HO-1信号通路发挥调节作用的。
Effect of ginsenoside Rg3 on oxidative stress injury of human lens epithelial cells
Objective To investigate the ameliorative effects of ginsenoside Rg3 on oxidative injury of human lens epithelial cells in-duced by hydrogen peroxide and the regulation of nuclear transcription factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods SRA01/04 cells induced by hydrogen peroxide were treated with different concentrations of ginsenosides Rg3,and the cell survival rate was measured by methyl thiazolyl tetrazolium(MTT)assay.The 3rd generation of logarithmic growth SRA01/04 cells were randomly divided into normal group,oxidative damage group(200 μmol∙mL-1 hydrogen peroxide treatment),low dose ginsenoside Rg3 group and high dose ginsenoside Rg3 group(40 μg∙mL-1 ginsenoside Rg3 treatment for 6 h,80 μg∙mL-1 gin-senoside RG3 treatment group,respectively.After the culture-medium was replaced and treated with 200 μmol∙mL-1 hydrogen pero-xide for 12 h),the cell survival rate was measured by MTT method,the cell apoptosis rate was measured by flow cytometry,kit was used to detect malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)content,The relative expression levels of Nrf2,Kelch like epichlorohydrin related protein 1(Keap1)and HO-1 were detected by Western blotting.Results Compared with 0 μg∙mL-1 ginsenoside Rg3 group,the cell survival rate of 10,20,40 and 80 μg∙mL-1 ginsenoside Rg3 group was gradually increased(P＜0.05).Compared with normal group,the cell survival rate,SOD and GSH-Px contents,relative expression levels of Nrf2,Keap1 and HO-1 proteins were decreased in oxidative injury group,while the cell apoptosis rate and MDA content were increased(P＜0.05).Compared with oxidative damage group,the cell survival rate,SOD and GSH-Px contents and relative expression levels of Nrf2,Keap1 and HO-1 proteins in low and high doses of ginsenoside Rg3 groups were increased,while the cell apoptosis rate and MDA content were decreased(P＜0.05).Ginsenoside Rg3 low dose group and ginsenoside Rg3 high dose group had the same changes in various indices,and ginsenoside Rg3 high dose group was more significant(P＜0.05).Conclusion Ginse-noside Rg3 inhibits H2O2-induced apoptosis of human lens epithelial cells and alleviates oxidative stress injury,possibly by activating Nrf2/HO-1 signaling pathway.
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