Abstract:

Abstract: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.

Key words: p38 mitogen-activated protein kinases;neurons;N-methylaspartate, rats,Spragne-Dawley;cell culture