p38MAPK蛋白在体外培养神经元中的表达及W-7的干预作用

目的:探讨p38MAPK蛋白在体外培养神经元中的激活及钙调蛋白抑制剂W-7的干预作用.方法:神经元原代培养并采用NSE免疫细胞化学技术鉴定神经元.取原代培养7 d的大鼠皮质神经元随机分为5组:正常对照组,仅用DMEM/F12完全培养基;NMDA组,去除正常神经元培养液,加入NMDA 50 μmol·L-1,处理时间10...

Expression of p38MAPK in cultured cortical neurons in vitro and intervention effect of W-7

Objective To investigate the activation of p38MAPK protein in cultured cortical neurons after NMDA injury and intervention effect of W-7.Methods The neurons were identified by polyclonal antibody against neuron specific enolase(NSE).The primary cortical neurons cultured for 7 d were randomly divided into 5 groups:control group;NMDA-injured group;three-doses of W-7 pretreatment groups.The cortical neurons were pre-cultured with regular media including different doses of W-7 respectively as 25,50 and 100 μmol·L-1 for 24 h before exposed to NMDA(50 μmol·L-1).The p38MAPK protein expression in cultured cortical neurons was detected by immunocytochemiscal staining and Western blotting.Results A large number of hippocampal neurons began to adhere to cover the glasses 6-12 h after culture.They showed different shapes after clinging to the plate.Their processes connected into nets and they were different in length and thickness.The proportion of positive neurons was 90.86%.Compared with control group,the expression of p38MAPK in cultured neuron was remarkably up-regulated in NMDA injured group by immunocytochemistrical staining and up-regulated in NMDA injured group by Western blotting(P<0.05),it was significantly down-regulated in W-7 pretreatment groups compared with NMDA group(P<0.05 or P<0.01).Conclusion NMDA could up-regulate the expressin of p38MAPK in cultured cotical neurons,and W-7 could down-regulated the expression of p38MAPK.