miR-383-5p转染对人卵巢颗粒细胞株KGN凋亡的影响及其机制

Effect and mechanism of miR-383-5p transfection on apoptosis of human ovarian granulosa cell line KGN

目的 观察miR-383-5p转染对人卵巢颗粒细胞株KGN凋亡的影响,并探讨其作用机制。方法 取对数生长期KGN细胞,分为3组,Blank组为空白对照组,miR-383-5p mimics组转染150 pmol的微小RNA-383-5p模拟物(miR-383-5p),miR-383-5p mimics+NAC组为转染150 pmol的miR-383-5p mimics后N-乙酰半胱氨酸(NAC)干预组,比较Blank组与miR-383-5p mimics组、miR-383-5p mimics组与miR-383-5p mimics+NAC组两组间凋亡率及凋亡相关蛋白(Bax、Bcl-2、cleaved caspase 3蛋白)的表达和活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)活性。结果 与Blank组比较,miR-383-5p mimics组凋亡率及Bax、cleaved caspase 3蛋白表达增加,Bcl-2蛋白表达减少(P均<0.05);与miR-383-5p mimics组比较,miR-383-5p mimics+NAC组miR-383-5p mimics组凋亡率及Bax、cleaved caspase 3蛋白表达减少,Bcl-2蛋白表达增加(P均<0.05)。与Blank组比较,miR-383-5p mimics组ROS、MDA水平升高,SOD活性降低(P均<0.05);与miR-383-5p mimics组比较,miR-383-5p mimics+NAC组ROS、MDA水平降低,SOD活性升高(P均<0.05)。结论 miR-383-5p过表达可促进KGN细胞凋亡,其作用机制可能是通过激活氧化应激途径。

Objective To investigate the effect of miR-383-5p transfection on apoptosis of human ovarian granulosacell line KGN,and to explore its mechanism.Methods KGN cells in the logarithmic growth phase were divided intothree groups. Blank group was the blank control group,KGN cells in the miR-383-5p mimics group were transfected with150 pmol of miR-383-5p mimics,and KGN cells in the miR-383-5p mimics+NAC group were added with NAC after cellswere transfected with 150 pmol of miR-383-5p mimics. The apoptosis rate,the expression level of apoptosis-related pro-teins Bax,Bcl-2 and cleaved caspase 3,reactive oxygen species(ROS)activity,MDA,and superoxide dismutase SODwere compared between Blank group and miR-383-5p mimics group,and between the miR-383-5p mimics group and miR-383-5p mimics+NAC group.Results Compared with the Blank group,the apoptosis rate increased significantly,theprotein expression of Bax and cleaved caspase 3 increased,and the expression of Bcl-2 protein decreased in the miR-383-5p mimics group(all P<0. 05). Further research showed that the apoptosis rate of cells in the miR-383-5p mimics+NACgroup was lower than that in the miR-383-5p group,the expression of Bcl-2 protein was higher,and the expression of Baxand cleaved caspase 3 protein was lower(all P<0. 05). Compared with the Blank group,the levels of ROS and MDA inthe miR-383-5p mimics group increased,and the activity of SOD decreased(all P<0. 05). Compared with the miR-383-5p mimics group,the ROS and MDA levels decreased,and the SOD activity increased in the miR-383-5p mimics+NAC group(all P<0. 05).Conclusion Overexpression of miR-383-5p can promote apoptosis of human ovarian granulosa cellline KGN probably by the activation of oxidative stress pathway.