The Effect Of Nogo-A On Phosphor-PKB Expression In PC12 Cells

Objective:To study the effect of Nogo-A on phospho-PKB expression in differentiated PC12 cells and investigate whether PKB was involved in the signaling pathway of neurite outgrowth that mediated by Nogo-A .Methods: Cultured cells were assigned to three groups, including control group1, control group2 and experimental group. Control group1:PC12 cells were routinely maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% new-born calf serum for 12 days. Control group2: PC12 cells were routinely maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% new-born calf serum and NGF(50 ng/ml). Ten days later, differentiated PC12 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) just containing 10% new-born calf serum for 2days. Experimental group: PC12 cells were routinely maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% new-born calf serum and NGF(50 ng/ml). Ten days later, differentiated PC12 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) just containing 10% new-born calf serum for 2days,and then the differentiated PC12 cells were treat with Nogo-A. There are two sub-groups in experimental group. Experimental group1: Nogo-A was added into the differentiated PC12 cells at a final concentration of 50nmol/L,100nmol/L,200nmol/L for 30 min. Experimental group2: Nogo-A was added into the differentiated PC12 cells at a final concentration of 200nmol/L for 10 min,30min 90min.The expression of phospho-PKB(Ser473) were probed by Western blots using corresponding antibodies.Results: The expression of phospho-PKB (Ser473) in differentiated PC12 cells treated by Nogo-A was markedly increased compared with control.Conclusions: this study indicated that Nogo-A may mediate the neurite outgrowth of PC12 cells in a PKB-dependent manner.