丹参酮IIA对过氧化氢诱导的人晶状体上皮细胞凋亡和氧化应激的影响  

Influence of tanshinone IIA on H2O2-induced apoptosis and oxidative stress in human lens epithelial cells via activating the Nrf2/HO-1 pathway

作　　者：蒋丽[1] 芮燕君 戴佳琪 丁高凤 JIANG Li;RUI Yanjun;DAI Jiaqi;DING Gaofeng(Aier Eye Hospital Affiliated to Central South University,Shanghai 200000,China)

机构地区：[1]中南大学附属爱尔眼科医院,上海市200000

出　　处：《眼科新进展》2021年第9期838-842,共5页Recent Advances in Ophthalmology

摘　　要：目的探讨丹参酮IIA(Tan IIA)对过氧化氢(H_(2)O_(2))诱导的人晶状体上皮细胞凋亡和氧化应激的调控作用及可能的机制。方法以人晶状体上皮细胞SRA01/04为研究对象,分为空白组、H_(2)O_(2)组(H_(2)O_(2)处理细胞建立氧化损伤模型)、Tan IIA组(Tan IIA溶液预处理24 h后,H_(2)O_(2)作用24 h)、Tan IIA+siNC组(转染阴性对照,其他处理方式同Tan IIA组)和Tan IIA+siNrf2组转染核因子E2相关因子2(Nrf2)siRNA(siNrf2),其他处理方式同Tan IIA组。CCK-8法检测细胞活力,流式细胞术分别检测细胞凋亡率和活性氧簇(ROS)水平,酶联免疫吸附试验测定细胞中过氧化氢酶(CAT)活性、超氧化物歧化酶(SOD)含量、谷胱甘肽过氧化物酶(GSH-Px)含量,Western blot检测细胞总蛋白中Nrf2、血红素氧合酶-1(HO-1)表达及核蛋白中Nrf2表达情况。结果通过CCK-8法确定H_(2)O_(2)和Tan IIA最佳实验浓度为300μmol·L^(-1)和20μmol·L^(-1)。与空白组相比,H_(2)O_(2)组SRA01/04细胞活力及细胞中CAT活性、SOD含量、GSH-Px含量均降低,凋亡率和ROS水平均增加,同时总蛋白中Nrf2和HO-1蛋白相对表达量及核蛋白中Nrf2蛋白相对表达量均下调(均为P<0.05)。与H_(2)O_(2)组相比,Tan IIA组SRA01/04细胞活力及细胞中CAT活性、SOD含量、GSH-Px含量均增加,凋亡率和ROS水平均降低,总蛋白中Nrf2和HO-1蛋白相对表达量及核蛋白中Nrf2蛋白相对表达量均上调(均为P<0.05)。而沉默Nrf2基因后,Tan IIA对SRA01/04细胞的保护作用被逆转。结论Tan IIA能够抑制H_(2)O_(2)诱导的人晶状体上皮细胞氧化应激性损伤和凋亡,其机制与Nrf2/HO-1通路的激活有关。Objective To study the effect of tanshinone IIA(Tan IIA)on H_(2)O_(2)-induced apoptosis and oxidative stress in human lens epithelial cells,and the possible mechanism.Methods The human lens epithelial cell line SRA01/04 was divided into blank group,H_(2)O_(2) group(treated with H_(2)O_(2)),Tan IIA group(pre-treated with 300μmol·L^(-1)Tan IIA solution for 24 h before H_(2)O_(2) exposure for 24 h),Tan IIA+siNC group(pre-treated with 300μmol·L^(-1)Tan IIA solution for 24 h before H_(2)O_(2) exposure for 24 h in cells transfected with siNC)and Tan IIA+siNrf2 group(pre-treated with 300μmol·L^(-1)Tan IIA solution for 24 h before H_(2)O_(2) exposure for 24 h in cells transfected with siNrf2).The cell viability was measured by CCK-8 assay.The apoptosis rate and the reactive oxygen species(ROS)level were measured by flow cytometry.Enzyme-linked immunosorbent assay(ELISA)was used to detect the activity of catalase(CAT)and the contents of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px).Western blot was used to detect protein levels of Nrf2 and heme oxygenase-1(HO-1).Results The optimal concentrations of H_(2)O_(2) and Tan IIA solution were determined by CCK-8 assay at 300μmol·L^(-1)and 20μmol·L^(-1),respectively.Compared with the blank group,the cell viability,CAT activity,and SOD and GSH-Px contents decreased,while the apoptosis rate and ROS level increased in SRA01/04 cells of H_(2)O_(2) group.Moreover,expression levels of total Nrf2 and HO-1,and nuclear protein level of Nrf2 were down-regulated(all P<0.05).Compared with H_(2)O_(2) group,the viability,CAT activity,and SOD and GSH-Px contents increased,while the apoptosis rate and ROS level decreased in SRA01/04 cells of Tan IIA group.Meanwhile,protein levels of total Nrf2 and HO-1,and nuclear protein level of Nrf2 were up-regulated(all P<0.05).The protective effect of Tan IIA on SRA01/04 cells was reversed by silencing Nrf2.Conclusion Tanshinone IIA can inhibit H_(2)O_(2)-induced apoptosis and oxidative stress in human lens epithelial cells via activ

关 键 词：丹参酮IIA 氧化应激 细胞凋亡 人晶状体上皮细胞 核因子E2相关因子2 血红素氧合酶-1 

分 类 号：R776[医药卫生—眼科]