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(P0.05),it was significantly down-regulated in W-7 pretreatment groups compared with NMDA group(P0.05 or P0.01).Conclusion NMDA could up-regulate the expressin of p38MAPK in cultured cotical neurons,and W-7 could down-regulated the expression of p38MAPK.