摘要
目的探讨人参皂苷Rg_(3)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Rg_(3)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200μmol∙mL^(−1)过氧化氢处理)、人参皂苷Rg_(3)低剂量组和人参皂苷Rg_(3)高剂量组(分别用40、80μg∙mL^(−1)人参皂苷Rg_(3)处理6 h,更换培养基后用200μmol∙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0μg∙mL^(−1)人参皂苷Rg_(3)组比较,10、20、40、80μg∙mL^(−1)人参皂苷Rg_(3)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GSHPx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Rg_(3)低剂量和人参皂苷Rg_(3)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Rg_(3)低剂量和人参皂苷Rg_(3)高剂量组各项指标水平变化规律相同,人参皂苷Rg_(3)高剂量组更显著(P<0.05)。结论人参皂苷Rg_(3)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。
Objective To investigate the ameliorative effects of ginsenoside Rg_(3) on oxidative injury of human lens epithelial cells induced 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 Rg_(3),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(200μmol∙mL^(−1) hydrogen peroxide treatment),low dose ginsenoside Rg_(3) group and high dose ginsenoside Rg_(3) group(40μg∙mL^(−1) ginsenoside Rg_(3) treatment for 6 h,80μg∙mL^(−1) ginsenoside RG3 treatment group,respectively.After the culture-medium was replaced and treated with 200μmol∙mL^(−1) hydrogen peroxide 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 0μg∙mL^(−1) ginsenoside Rg_(3) group,the cell survival rate of 10,20,40 and 80μg∙mL^(−1) ginsenoside Rg_(3) 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 Rg_(3) groups were increased,while the cell apoptosis rate and MDA content were decreased(P<0.05).Ginsenoside Rg_(3) low dose group and ginsenoside Rg_(3) high d
作者
张可
李莉
ZHANG Ke;LI Li(Department of Ophthalmology,Yanling Hospital of Traditional Chinese Medicine,Xuchang City,He’nan Province,Xuchang 461200,China;Department of Ophthalmology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《西北药学杂志》
2024年第1期55-60,共6页
Northwest Pharmaceutical Journal
基金
河南省2019年科技发展计划项目(编号:192102310341)。
