Midazolam inhibits hypoxia/reoxygenation-induced PC12 cell apoptosis through up-regulating miR-320
Objective In order to clarify the role of midazolam in ischemic stroke,the effect of midazolam on hypoxia/reoxygenation(H/R)-induced proliferation and apoptosis of PC12 cells and its potential mechanism were investigated.Methods PC 12 cells were cultured in vitro and divided into control group,H/R group,midazolam group with different doses,10 µmol/L midazolam+anti-miR-NC group,and 10 μmol/L midazolam+anti-miR-320 group.Cell proliferation and apoptosis were detected by CCK-8 method and flow cytometry;Protein expression was tested by Western blot;miR-320 expression was examined by RT-qPCR.Results Compared with the control group,cell A value(0.43±0.02),pro-caspase3 level(0.14±0.01)and miR-320 expression(0.15±0.01)were signif-icantly decreased(P＜0.05),while apoptosis rate(26.68±0.81)and Cleaved-caspase3 level(0.70±0.05)were sig-nificantly increased in the H/R group(P＜0.05).Compared with the H/R group,cell A value(0.43±0.02,0.57± 0.03,0.93±0.04),pro-caspase3 level(0.14±0.01,0.30±0.02,0.52±0.03)and miR-320 expression(0.15±0.02,0.42±0.03,0.81±0.05)were remarkably enhanced(P＜0.05),while apoptosis rate(25.65±0.92,21.74±0.56,15.63±0.50)and cleaved-caspase3 level(0.70±0.06,0.50±0.04,0.23±0.02)were significantly reduced in the mid-azolam group with different doses(0.1,1.0,10 µmol/L)(P＜0.05),in a dose-dependent manner.Compared with the 10 μmol/L midazolam+anti-miR-NC group,cell A value(0.47±0.02),pro-caspase3 level(0.24±0.01)and miR-320 expression(0.26±0.02)were markedly inhibited(P＜0.05),while apoptosis rate(23.52±0.59)and Cleaved-caspase3 level(0.56±0.04)were remarkably promoted in the 10 μmol/L midazolam+anti-miR-320 group(P＜0.05).Conclusion Midazolam may upregulate miR-320 expression to promote H/R-induced PC 12 cell proliferation and in-hibit apoptosis.
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