咪达唑仑调节RhoA/ROCK信号通路对OGD/R诱导的神经元凋亡的影响OA
Effect of midazolam on OGD/R-induced neuronal apoptosis by regulating RhoA/ROCK signaling pathway
目的:探讨咪达唑仑(MDZ)调节Rho蛋白相关卷曲螺旋激酶(RhoA)/Rho激酶(ROCK)信号通路对氧糖剥夺再复氧(OGD/R)诱导的神经元凋亡的影响.方 法:HT22细胞分为Control组(正常培养)、OGD/R组(进行OGD/R诱导)、L-MDZ、M-MDZ、H-MDZ组(3、6、12 μmol/mL MDZ预处理2 h,进行OGD/R诱导)、MDZ+溶血磷脂酸(LPA)(12 μmol/mL MDZ和50 μmol/L LPA预处理2 h,进行OGD/R诱导)组.MTT法和平板克隆法检测MDZ对HT22细胞增殖的影响;流式细胞术检测MDZ对HT22细胞凋亡的影响;ELISA检测MDZ对HT22细胞炎症因子(TNF-α、IL-1β、IL-6)和氧化应激(ROS、MDA、SOD)指标表达的影响;Western blot检测HT22细胞Bax、RhoA、ROCK、Bcl-2蛋白表达.结果:OGD/R诱导导致HT22细胞存活率、克隆数下降,凋亡率上升(P<0.05);L-MDZ、M-MDZ、H-MDZ处理后存活率、克隆数上升,凋亡率下降(P<0.05);MDZ+LPA处理后存活率、克隆数下降,凋亡率上升(P<0.05).OGD/R组SOD、Bcl-2蛋白表达低于Control组,TNF-α、IL-1β、IL-6、ROS、MDA、Bax、RhoA、ROCK蛋白表达高于Control组(P<0.05);L-MDZ组、M-MDZ组、H-MDZ组SOD、Bcl-2蛋白表达高于OGD/R组,TNF-α、IL-1β、IL-6、ROS、MDA、Bax、RhoA、ROCK蛋白表达低于OGD/R组(P<0.05);MDZ+LPA组SOD、Bcl-2蛋白表达低于H-MDZ组,TNF-α、IL-1β、IL-6、ROS、MDA、Bax、RhoA、ROCK蛋白表达高于H-MDZ组(P<0.05).结论:MDZ可抑制OGD/R诱导的神经元凋亡,其机制可能是通过抑制RhoA/ROCK信号通路实现的.
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.
赵晨;卫聪慧;张迅
滕州市中心人民医院麻醉科,滕州 277588滕州市中心人民医院病理科,滕州 277588滕州市中心人民医院麻醉科,滕州 277588
医药卫生
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
MidazolamRho associated coiled coil containing protein kinaseRho kinaseOxygen glucose deprivation and reoxygenationNeuronal apoptosis
《中国免疫学杂志》 2026 (5)
1084-1088,5
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