Protective Effect Of Apelin Against Glutamate-Induced Neurotoxicity To PC12 Cells By Modulation Of ERK Pathway

Objective To observe effect of Apelin-13 on PC12 cell injury induced by glutamate and to investigate the mechanism which Apelin-13 protect against the glu tamate-induced injury of neurotoxity from the aspect of ERK1/2 signalin g pathwayMethodsThe modle of glutamate-induced injury in PC12 cells was established.The cells were incubated by different glutamate concentration(5、10、20 mmol/L) to find the best concentration of damage;The cells was incubated by different Apelin-13 concentration(10、100nmol/l;1、10μmol/L) with glutamate(10 mmol/L) to find the best concentration of protection.;Then U0126(ERK1/2 inhibitors) was added and the experiment is divided into 5 groups:Control、Glu、Glu+Apelin-13、Glu+Apelin-13+U0126 、 U0126,U0126 was pretreated 30 min,then common cultureed with Apenlin-13 and glutamate.The viability of PC12 cell was detected by MTT method;The apoptosis of PC12 cells was detected by Flow cytometry(FCM);The expressions of p-ERK1/2 was determined by Western Blot. Results 1. The injury of PC12 cells was induced by glutamate.Treatment with glutamate(5,10,20 mmol/L, 24 h) attenuated cell viability;increased cell apoptosis ratio and the expressions of p-ERK1/2 were decreased in a dose-dependent manner. 2. The damage of PC12 cell induced by glutamate was reversed via Apelin-13.Apelin-13(1 μmol/L)and glutamate(10 mmol/L) co-treated PC12 cell for 24 h,the cell viability increased significantaly,the apopotosis ratio reduced significantly.That shows Apelin-13 against the neurotoxity effects of glutamate on PC 12 cells. 3.ERK patheway mediates the protective effects of Apelin against glutamate-induced neurotoxicityPC12 cells were pretreated by U0126(ERK1/2 inhibitor) for 30 min before incubating with Apelin-13 and glutamate.The protective effect of Apelin-13 on PC12 cell injury induced by glutamate was blocked.Compared with the Apelin-13 and glutamate group,the cell viability was decreased,the cell apoptosis was increased,the expression of p-ERK1/2 was decreased obviously in U0126 group. Conclusion:Apelin-13 plays a protective role on the PC12 cell demage induced by glutamate,in which ERK1/2 signaling pathways involved.