Effects of miR-27a-3p on neuronal apoptosis induced by oxygen glucose deprivation/reoxygenation via tar-geting Rnd3

LI Junjie; BAI Wenya; CHEN Wendong; YANG Wei; SHAO Jianlin

doi:10.3969/j.issn.1006-5725.2023.23.005

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.