P38MAPK信号通路对缺血预处理减轻神经元凋亡作用机制的研究  

Role of P38MAPK Signaling Pathway on Preconditioning-induced Apoptosis in Cortical Neurons

作　　者：李昕华[1] 刘艳华[1] 蔺勇[1] LI Xinhua;LIU Yanhua;LIN Yong(Changchun Central Hospital,Jilin 130051,China)

机构地区：[1]长春市中心医院,吉林长春130051

出　　处：《北华大学学报（自然科学版）》2020年第6期744-747,共4页Journal of Beihua University（Natural Science）

基　　金：吉林省卫生厅科研课题(2011Z015).

摘　　要：目的探讨通过缺血预处理抑制P38丝裂素活化蛋白激酶(P38MAPK)信号通路以减轻神经元凋亡的机制.方法将原代培养的神经元及星形胶质细胞进行缺血预处理,将常规条件下培养的星形胶质细胞培养基作为条件培养基1(ACM1);在分泌GDNF最多时,将星形胶质细胞培养基制备成条件培养基2(ACM2);GDNF基因经转染质粒沉默后提取其培养基,作为条件培养基3(ACM3),将P38MAPK的抑制剂SB203580加入ACM3中,制备成ACM4.将对照组、预处理组、预处理+缺血组、缺血组神经元孵育在不同条件培养基中,AO/EB荧光染色检测神经元凋亡情况,Western blot法检测神经元p-P38MAPK及Caspase-3的表达.结果预处理+缺血组和缺血组神经元凋亡率明显增高(P<0.05),加入ACM2后神经元凋亡较ACM1和ACM3两组均减少(P<0.05).预处理+缺血组和缺血组凋亡的神经元明显多于另外两组(P<0.05),加入ACM1和ACM3两种培养基后凋亡率高于ACM2组(P<0.05).预处理+缺血组及缺血组p-P38MAPK及Caspase-3的蛋白表达均明显高于对照组和预处理组(P<0.05),预处理+缺血组p-P38MAPK及Caspase-3的蛋白表达均较缺血组低(P<0.05),加入ACM2组的p-P38MAPK及Caspase-3蛋白表达低于另外两组(P<0.05).结论星形胶质细胞经过缺血预处理后GDNF分泌增多,通过抑制P38MAPK信号通路及Caspase-3的表达减少神经元凋亡,进而发挥内源性神经保护机制.Objective To study the system of ischemic preconditioning to reduce neuron apoptosis by inhibiting P38MAPK signaling pathway.Method The primary cultured neurons and astrocytes were pretreated with ischemia.The normal astrocyte medium were prepared into ACM1;The astrocytes medium with the highest GDNF secretion were prepared into ACM2;After the GDNF gene was silenced by the transfected plasmid,the culture medium was extracted as ACM3.Neurons were divided into control group,preconditioning group,preconditioning+ischemia group and ischemia group.The neurons were incubated in different ACM,the apoptosis of neurons were detected by AO/EB double staining technique.The level of p-P38MAPK in neurons were detected by Western blot.The protein expressions of Caspase-3 were detected by using Western blot.Results The apoptotic rate of neurons in preconditioning+ischemia group and ischemia group were significantly higher(P<0.05).The apoptotic rate of neurons in ACM2 group was lower than those in ACM1 and ACM3 group(P<0.05).The expression of p-P38MAPK and Caspase-3 in the preconditioning+ischemia group and ischemia group were significantly higher than those in the control group and preconditioning group(P<0.05),the expression of p-P38MAPK and Caspase-3 in the preconditioning+ischemia group was lower than that in the ischemia group(P<0.05).The expression of p-P38MAPK and Caspase-3 in the ACM2 group were less than those of other two groups(P<0.05).Conclusion GDNF secretion of astrocytes increased after ischemic preconditioning,and decreased neuron apoptosis by inhibiting P38MAPK signaling pathway and Caspase-3 expression,playing an endogenous protective mechanism.

关 键 词：P38丝裂素活化蛋白激酶 半胱氨酸天冬酶-3 缺血预处理 神经元 凋亡 信号通路 

分 类 号：R34[医药卫生—基础医学]