摘要
目的:探讨尼莫地平(NM)调节受体相互作用蛋白激酶1(RIP1)/受体相互作用蛋白激酶3(RIP3)/混合谱系激酶样结构域蛋白(MLKL)信号通路对氧糖剥夺/复氧(OGD/R)诱导的神经元坏死性凋亡的影响。方法:HT22细胞分为Control组、OGD/R组、L-NM组、M-NM组、H-NM组、Z-VAD-fmk组。CCK-8法检测HT22细胞增殖;流式细胞术检测HT22细胞凋亡;DCFH-DA荧光探针检测HT22细胞中ROS水平;ELISA检测HT22细胞中炎症因子(IL-6、TNF-α、IL-1β)表达;Western blot检测HT22细胞中RIP1、RIP3、MLKL蛋白表达。结果:OGD/R组HT22细胞存活率低于Control组,细胞凋亡率、ROS阳性细胞比例、IL-6、TNF-α、IL-1β、RIP1、RIP3、MLKL蛋白表达高于Control组(P<0.05);L-NM组、M-NM组、H-NM组细胞存活率高于OGD/R组,细胞凋亡率、ROS阳性细胞比例、IL-6、TNF-α、IL-1β、RIP1、RIP3、MLKL蛋白表达低于OGD/R组(P<0.05);Z-VAD-fmk组细胞存活率低于H-NM组,细胞凋亡率、ROS阳性细胞比例、IL-6、TNF-α、IL-1β、RIP1、RIP3、MLKL蛋白表达高于H-NM组(P<0.05)。结论:NM可抑制OGD/R诱导的神经元坏死性凋亡,其机制可能通过抑制RIP1/RIP3/MLKL信号通路实现。
Objective:To investigate effect of nimodipine(NM)on neuronal necrotic apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)by regulating receptor-interacting protein kinase 1(RIP1)/receptor-interacting protein kinase 3(RIP3)/mixed lineage kinase-like domain(MLKL)signaling pathway.Methods:HT22 cells were separated into control group,OGD/R group,L-NM,M-NM,H-NM groups and Z-VAD-fmk group.CCK-8 method was used to detect HT22 cell proliferation.Flow cytometry was used to detect apoptosis of HT22 cells.DCFH-DA fluorescent probe was used to detect ROS level in HT22 cells.ELISA was used to detect expressions of inflammatory factors(IL-6,TNF-α,IL-1β)in HT22 cells.Western blot was used to detect protein expressions of RIP1,RIP3 and MLKL in HT22 cells.Results:Survival rate of HT22 cells in OGD/R group was lower than control group,apoptosis rate,proportion of ROS-positive cells,expressions of IL-6,TNF-α,IL-1β,RIP1,RIP3 and MLKL proteins were higher than control group(P<0.05).Survival rate in L-NM,M-NM and H-NM groups were higher than OGD/R group,apoptosis rate,proportion of ROSpositive cells,expressions of IL-6,TNF-α,IL-1β,RIP1,RIP3 and MLKL proteins were lower than OGD/R group(P<0.05).Survival rate in Z-VAD-fmk group was lower than H-NM group,apoptosis rate,proportion of ROS-positive cells,expressions of IL-6,TNF-α,IL-1β,RIP1,RIP3 and MLKL proteins were higher than H-NM group(P<0.05).Conclusion:NM can inhibit OGD/R-induced neuronal necroptosis,whose mechanism may be achieved by inhibiting RIP1/RIP3/MLKL signaling pathway.
作者
刘冰
周春秀
叶爽
LIU Bing;ZHOU Chunxiu;YE Shuang(Department of Neurology,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China;Department of Critical Care Medicine,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China)
出处
《中国免疫学杂志》
北大核心
2026年第2期311-315,共5页
Chinese Journal of Immunology
基金
河南省医学科技攻关计划项目(LHGJ20220812)。
