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The Synergistic Protective Effect and Mechanism of NGF and Ganglionside GM1 on the Intoxication of 2,5-Hexanedione in PC12 Cells

Author: LiuShaoPing
Tutor: YangGuoCheng
School: Wuhan University
Course: Biochemistry and Molecular Biology
Keywords: Nerve Growth Factor Ganglioside GM1 2,5 - dione Protective Effect of Combined Phosphatidylinositol 3 - OH kinase Akt
CLC: R363
Type: Master's thesis
Year: 2005
Downloads: 67
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Abstract


Objective: To establish a rat adrenal pheochromocytoma PC12 cells 2,5 - hexanedione (2,5-HD) poisoning model studies of nerve growth factor (Nerve Growth Factor, NGF) and ganglioside GM1 ( Ganglioside GM1 ) and to explore its possible mechanism of action of the protective effect of 2,5-HD intoxication . Method : the cells were randomly divided into a control group and the experimental group and control group in the growth of normal medium , the experimental group dealt with different concentrations of 2,5- HD , NGF and GM1 and 50μmol / L PI3- K inhibitor LY294002 . Giemsa staining in cell morphology was observed under an inverted microscope , MTT assay of cell viability and apoptosis rate of flow cytometry assay , immunoblotting to detect the protein expression of Akt and phosphorylated -Akt (P-Akt) . Results: With the concentration of 2,5-HD processing NGF and GM1 pretreatment of the cells increased and prolonged cell injury aggravated reduced survival ( P lt; 0.05 ) , the rate of apoptosis increased ( P lt; 0.05 ) ; show significant resistance to damage , improve cell viability (P lt; 0.05), apoptosis rate ( P lt; 0.001) , and more significant than a separate application effect in the United ( P lt; 0.001 ) ; of NGF was good dose-dependent protection of PC12 cells ( P lt; 0.001 ) GM1 Whether used alone or used in combination , are 50μmol / L role best . Western blot NGF and GM1 can promote the phosphorylation of Akt expression in a dose - dependent manner , the effect is particularly evident when used in combination , had no significant effect on Akt level ; signal blocking PI3-K PI3-K inhibitor LY294002 pretreatment NGF and GM1 -induced Akt phosphorylation was almost completely blocked . Conclusion :2,5-HD dose- and time -dependent inhibition of PC12 cell survival and proliferation , induction of fine damage and apoptosis . 2,5-HD toxicity in PC12 cells , NGF and GM1 showed a synergistic protective effect . This protective effect could be generated by starting the PI3-K signaling pathway , activation of Akt and its downstream signaling .

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