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研究MPP+(1-methyl-4-phenylpyridinium)对PC12细胞的毒性作用及机制。采用MTT法检测细胞存活率,Western blotting检测蛋白变化及双荧光素酶报告基因系统检测转录因子活性。结果表明,MPP+能够显著抑制细胞存活率,且呈剂量依赖性;而且MPP+能够降低PC12细胞神经营养因子BDNF(brain-derived neurotrophic factor)的蛋白水平,并能够抑制ERK(extracellular signal-regulated proteinkinase)的磷酸化,而对CaMKⅡ活性无影响,同时能够显著抑制BDNF的转录因子CREB(cAMP response element binding protein)的磷酸化,并抑制其转录活性。因此MPP+可能通过抑制ERK活性而抑制其下游转录因子CREB的活性,最终导致BDNF的表达减弱。
To study the toxic effect and mechanism of MPP + (1-methyl-4-phenylpyridinium) on PC12 cells. Cell viability was detected by MTT assay, protein changes were detected by Western blotting and transcription factor activity was detected by dual luciferase reporter system. The results showed that MPP + could significantly inhibit cell viability in a dose-dependent manner. Moreover, MPP + could reduce the protein level of brain-derived neurotrophic factor (BDNF) in PC12 cells and inhibit the extracellular signal-regulated protein kinase Phosphorylation, but had no effect on the activity of CaMKII, at the same time it could significantly inhibit the phosphorylation of cAMP response element binding protein (CREB) of BDNF and inhibit its transcriptional activity. Therefore, MPP + may inhibit the activity of its downstream transcription factor CREB by inhibiting the activity of ERK, leading to the decrease of BDNF expression.