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Protective Effect Of Mouse 2.5s Nerve Growth Factor To The Intoxication Of PC12 Cell Induced By 2,5-hexanedione

Posted on:2006-06-11Degree:MasterType:Thesis
Country:ChinaCandidate:L C SunFull Text:PDF
GTID:2144360182467077Subject:Biochemistry and Molecular Biology
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Objective To build the model of injury and apoptosis of PC12 induced by 2,5-hexanedone(2,5-HD). And then to explore that if the nerve growth factor(NGF) could have protective effects on the injury of PC12 induced by 2,5-HD. Methods With differentiated PC12 cell as the model of neurons, different concentrations of 2,5-HD and NGF were added into the culture of PC12 cells, and then cell viability was tested with MTT; change of nucleolus was observed with fluoroscope; DNA fragment was observed with agarose gel electrophoresis; apoptosis ratio was tested with flow cytometry (FACS); p53 protein , NF-H protein, phospho-NF-H protein, ERK1/2 protein and phospho-ERK1/2 protein was detected with western blot; and the differences between each groups were compared. Results Cell viabilities decreased with the concentrations of 2,5-HD increasing(P<0.05). Hochest33258 staining could distinguish apoptotic cells as well as normal cells. There were obvious DNA fragment by agarose gel electrophoresis. The apoptosis ratio and the concentration of p53 protein increased with the concentrations of 2,5-HD getting higher.(P<0.05).There were obvious phospho-ERK1/2 protein after 2,5-HD were added. But cell viabilities increased with the concentrations of NGF increasing(P<0.05). DNA fragment, the apoptosis ratio, the concentration of p53 and the ratio of phospho-NF-H protein to NF-H protein all decreased with the concentrations of NGF getting higher.(P<0.05). Conclusion 2,5-HD could injury PC12 cell and induce it to apoptosis. And NGF might have direct nutrition effects to PC12 cells, and protect them from injury induced by 2,5-HD.
Keywords/Search Tags:nerve growth factor, 2,5-hexanedione, apoptosis, PC12
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