实用医学杂志2023,Vol.39Issue(23):3051-3057,7.DOI:10.3969/j.issn.1006-5725.2023.23.005
miR-27a-3p靶向Rnd3对氧糖剥夺/复氧诱导的神经细胞凋亡的影响
Effects of miR-27a-3p on neuronal apoptosis induced by oxygen glucose deprivation/reoxygenation via tar-geting Rnd3
摘要
Abstract
Objective To investigate the effect and mechanism of miR-27a-3p on nerve cell apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)through regulation of Rho family GTPase 3(Rnd3)expression.Methods PC12 neurons were cultured in vitro and reoxygenated for 3,6,9 h and 12 h after 2 h oxygen glucose deprivation.Cell viability,miR-27a-3p expression and Rnd3 mRNA expression were assessed at each time point and the optimal reoxygenation time point was screened.After transfection of miR-27a-3p Mimic,miR-27a-3p Inhibitor and their negative control,transfection of shRnd3 and its negative control,or co-transfection of shRnd3 and miR-27a-3p Inhibitor through lentivirus,CCK-8 assay was used to detect cell activity.The apoptosis rate of the cells was detected using flow cytometry.Expression of miR-27a-3p and Rnd3 mRNA was detected by RT-qPCR.Expression of apoptosis-related protein and Rnd3 protein was detected by Western blot.The dual luciferase reporter assay confirmed the targeting relationship between miR-27a-3p and Rnd3.Results Upregulation of miR-27a-3p increased cell viability,decreased total cell apoptosis rate,suppressed pro-apoptotic proteins Cleaved Caspase-3(C-caspase-3)and Bax,and promoted expression of anti-apoptotic protein Bcl-2(P<0.05);The opposite result was found when down-regulating miR-27a-3p.The double luciferase reporter gene assay showed that Rnd3 was the target gene of miR-27a-3p.Down-regulation of Rnd3 increased cell viability,decreased the total rate of apoptosis,suppressed the pro-apoptotic protein C-caspase-3,Bax,and promoted expression of the anti-apoptotic protein Bcl-2(P<0.05).However,miR-27a-3p Inhibitor reversed the protective effect of shRnd3.Conclusion miR-27a-3p alleviates OGD/R-induced damage to PC12 neurons by targeting Rnd3 to inhibit cell apoptosis.关键词
miR-27a-3p/氧糖剥夺/复氧/Rnd3/细胞凋亡Key words
miR-27a-3p/OGD/R/Rnd3/cell apoptosis分类
医药卫生引用本文复制引用
李俊杰,白文娅,陈文栋,杨伟,邵建林..miR-27a-3p靶向Rnd3对氧糖剥夺/复氧诱导的神经细胞凋亡的影响[J].实用医学杂志,2023,39(23):3051-3057,7.基金项目
国家自然科学基金地区基金项目(编号:81760248) (编号:81760248)
云南省科技计划重点项目(编号:2018FA042) (编号:2018FA042)
云南省基础研究计划项目(编号:202101AY070001-116) (编号:202101AY070001-116)