Abstract
Objective:To investigate effect of midazolam(MDZ)on neuronal apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)by regulating Rho associated coiled coil containing protein kinase(RhoA)/Rho kinase(ROCK)signaling pathway.Methods:HT22 cells were divided into Control group(normal culture),OGD/R group(OGD/R induction),L-MDZ,M-MDZ,H-MDZ groups(3,6,12 μmol/mL MDZ pretreatment for 2 h,OGD/R induction)and MDZ+lysophosphatidic acid(LPA)group(12 μmol/mL MDZ and 50 μmol/L LPA pretreatment for 2 h for OGD/R induction).MTT and plate cloning were used to detect effect of MDZ on proliferation of HT22 cells.Flow cytometry was used to detect effect of MDZ on apoptosis in HT22 cells.ELISA was used to detect effect of MDZ on expressions of inflammatory cytokines(TNF-α,IL-1β,IL-6)and oxidative stress(ROS,MDA,SOD)in HT22 cells.Western blot was used to detect expressions of Bax,RhoA,ROCK and Bcl-2 proteins in HT22 cells.Results:After OGD/R induction,survival rate and clone number of HT22 cells were decreased,while apoptosis rate was increased(P<0.05).Survival rate and clone number of L-MDZ,M-MDZ and H-MDZ treatments were increased,while apoptosis rate was decreased(P<0.05).After MDZ+LPA treatment,survival rate and clone number were decreased,while apoptosis rate was increased(P<0.05).SOD and Bcl-2 protein expression of HT22 cells in OGD/R group were lower than control group,TNF-α,IL-1β,IL-6,ROS,MDA,Bax,RhoA and ROCK proteins were higher than control group(P<0.05).SOD and Bcl-2 protein expressions in L-MDZ group,M-MDZ group and H-MDZ group were higher than OGD/R group,TNF-α,IL-1β,IL-6,ROS,MDA,Bax,RhoA and ROCK proteins were lower than OGD/R group(P<0.05).SOD and Bcl-2 protein expression in MDZ+LPA group were lower than H-MDZ group,TNF-α,IL-1β,IL-6,ROS,MDA,Bax,RhoA and ROCK proteins were higher than H-MDZ group(P<0.05).Conclusion:MDZ can inhibit OGD/R induced neuronal apoptosis,and its mechanism may be achieved by inhibiting RhoA/ROCK signaling pathway.关键词
咪达唑仑/Rho蛋白相关卷曲螺旋激酶/Rho激酶/氧糖剥夺再复氧/神经元凋亡Key words
Midazolam/Rho associated coiled coil containing protein kinase/Rho kinase/Oxygen glucose deprivation and reoxygenation/Neuronal apoptosis分类
医药卫生
