miR-630对H2O2诱导的人晶状体上皮细胞的增殖、凋亡与氧化损伤的影响  

Effects of miR-630 on H2O2-induced Proliferation,Apoptosis and Oxidative Damage of Human Lens Epithelial Cells

作　　者：周洋[1] 陈婷妍 美丽巴努·玉素甫[1] Zhou Yang;Chen Tingyan;Meilibanu·Yusufu(Ophthalmology Department,Fifth Affiliated Hospital of Xinjiang Medical University,Xinjiang 830011,China)

机构地区：[1]新疆医科大学第五附属医院眼科,乌鲁木齐830011 [2]成都华厦眼科医院小儿斜弱视科,610041

出　　处：《医学研究杂志》2020年第6期84-90,共7页Journal of Medical Research

基　　金：新疆维吾尔自治区自然科学基金资助项目(2019D01C271)。

摘　　要：目的探讨miR-630对H2O2诱导的人晶状体上皮细胞的增殖、凋亡与氧化损伤的影响。方法将HLE-B3细胞分为对照组、H2O2组、H2O2+阴性对照组(H2O2+miR-NC)和H2O2+miR-630抑制剂组(H2O2+miR-630 inhibitor)。qRT-PCR检测各组细胞中miR-630的表达水平。细胞集落形成实验和EdU实验分析各组细胞的增殖能力。细胞划痕愈合实验分析各组细胞的迁移能力。AnnexinⅤ-FITC/PI分析各组细胞凋亡能力。试剂盒检测ROS水平和SOD含量。Western blot法检测各组细胞中Bcl-2和Akt蛋白的表达。结果与对照组比较,H2O2组和H2O2+miR-NC组中miR-630的表达显著上调(P=0.000);与H2O2组和H2O2+miR-NC组比较,低表达miR-630使HLE-B3细胞中miR-630的表达降低。与对照组比较,H2O2组和H2O2+miR-NC组HLE-B3的增殖能力(P均<0.000)、迁移能力显著下降(P均=0.000),凋亡能力明显提高(P均=0.000);与H2O2组和H2O2+miR-NC组比较,H2O2+miR-630 inhibitor组HLE-B3细胞的增殖能力(P均<0.01)、迁移能力提高(P均<0.05),凋亡能力显著下降(P=0.000)。与对照组比较,H2O2组和H2O2+miR-NC组中ROS水平上调(P均=0.000)、SOD含量减少(P=0.000)、Bcl-2和Akt蛋白表达水平下调(P均=0.000);与H2O2组和H2O2+miR-NC组比较,H2O2+miR-630 inhibitor组中ROS水平下调(P均<0.01)、SOD含量增加(P<0.05)、Bcl-2和Akt蛋白表达水平上调(P<0.01,P<0.05)。结论低表达miR-630可能促进H2O2诱导的人晶状体上皮细胞的细胞增殖、迁移,抑制细胞凋亡和减少氧化损伤,其可能与Bcl-2和Akt的表达上调相关。Objective To investigate the effect of miR-630 on H2O2-induced proliferation,apoptosis and oxidative damage of human lens epithelial cells.Methods HLE-B3 cells were divided into control group,H2O2 group,H2O2+negative control group(H2O2+miR-NC)and H2O2+miR-630 inhibitor group(H2O2+miR-630 inhibitor).qRT-PCR was used to detect the expression level of miR-630 in each group of cells.Cell colony formation assay and EdU assay were used to analyze the proliferative capacity of each group of cells.Cell scratch healing experiments were performed to analyze the migration ability of each group of cells.AnnexinⅤ-FITC/PI was used to analyze the apoptosis ability of each group.The kit was used to detect ROS levels and SOD levels.The expression of Bcl-2 and Akt protein in each group was detected by Western blot.Results Compared with the control group,the expression of miR-630 was significantly up-regulated in the H2O2 group and the H2O2+miR-NC group(P=0.000).Low expression of miR-630 reduced the expression of miR-630 in HLE-B3 cells compared to the H2O2 and H2O2+miR-NC groups.Compared with the control group,the proliferation ability(both P=0.000)and migration ability of HLE-B3 in H2O2 group and H2O2+miR-NC group were significantly decreased(both P=0.000),and the apoptosis ability was significantly improved(all P=0.000).Compared with the H2O2 group and the H2O2+miR-NC group,the proliferation ability(P<0.01)and migration ability of HLE-B3 cells in H2O2+miR-630 inhibitor group(both P<0.05),and the apoptosis ability decreased significantly(P=0.000).Compared with the control group,ROS levels were up-regulated(both P=0.000),SOD levels were decreased(P=0.000),and Bcl-2 and Akt protein levels were down-regulated in H2O2 and H2O2+miR-NC groups(both P=0.000).Compared with H2O2 group and H2O2+miR-NC group,ROS levels were down-regulated(both P<0.01),SOD content increased(P<0.05),and Bcl-2 and Akt protein expression levels were up-regulated in H2O2+miR-630 inhibitor group(P<0.01,P<0.05).Conclusion Low expression of miR-630 may promote H2O

关 键 词：miR-630 H2O2 人晶状体上皮细胞 增殖 凋亡 氧化损伤 

分 类 号：R587.2[医药卫生—内分泌] R747.9[医药卫生—内科学]