The Synergistic Protective Effect And Mechanism Of NGF And Ganglionside GM1 On The Intoxication Of 2,5-Hexanedione In PC12 Cells

Object: To establish a model of 2,5-hexanedione (2,5-HD) intoxication in PC12 cells. To investigate whether NGF and GMl have synergistic protective effect on 2,5-HD intoxication in PC12 cells and potential mechanism involved.Methods: Cells were divided into control group and treated groups at random. Control group was maintained in normal medium; treated groups were exposed to different concentrations of 2,5-HD with or without NGF, GMl and 50 umol/L LY294002. The cells' morphological changes were observed by inverted microscope after being dyed with Giemsa. Cell viability was determined by MTT assay. The apoptotic ratio was determined by flow cytometry (FCM). The expression of Akt and P-Akt were detected by immunoblotting.Results: With the concentration of 2,5-HD increasing and treated time extension, PC12 cells injury aggravated, the cell viability decreased (P<0.05), and the apoptotic ratio increased(P<0.05). PC12 cells pretreated with NGF and GMl obviously resisted injury, the cell viability increased (P<0.05), apoptotic ratio decreased (P<0.001). Combination of NGF and GMl afforded additional protection that was greater than that of either agent individually (P<0.01), NGF showed protective effect in a dose-dependent manner. 50 umol/L GMl showed the most evident protection in GMl groups. Immunoblotting showed that NGF and GMl had no effect on Akt exprssion, yet they could increase the expression of phosph-Akt, especially when in combination, which would be blocked by LY294002 (inhibitor of PI3-K).Conclusion: 2,5-HD inhibited the survival, induced apoptosis and injury of PC12 cells in a dose- and time-dependent manner. NGF and GMl showed synergistic protective effect on 2,5-HD intoxication in PC12 cells. This effect may rely on the activation of PI3-K/Akt pathway.