【摘要】 目的:探讨H2O2对PC12细胞par-4和caspase-3基因表达的影响,为阿尔茨海默病神经元凋亡发生的机制提供实验依据。方法:用不同剂量H2O2(0～400nmol/L)处理PC12细胞8h,或终浓度200nmol/LH2O2处理PC12细胞不同时间(0～8h),采用RT-PCR检测par-4,caspase-3mRNA表达变化,Westernblot检测Par-4蛋白表达和Caspase-3P20活性片段的变化,用比色法检测Caspase-3相对活性。结果:随H2O2浓度从100～400nmol/L增加,par-4,caspase-3mRNA表达和Par-4蛋白表达水平及Caspase-3P20活性片段逐渐增加;200nmol/LH2O2作用PC12细胞0～8h,par-4mRNA表达和Par-4蛋白表达,在2～8h随时间延长表达增加;caspase-3mRNA表达和Caspase-3P20活性片段,在4～8h随时间延长而增加;随H2O2浓度从50～400nmol/L增加,Caspase-3相对活性增加(χ2=4.8,P<0.05),在400nmol/LH2O2时Caspase-3相对活性达最大值。结论:随着H2O2剂量依赖性的增加和同一剂量下H2O2处理PC12细胞时间增加par-4,caspase-3基因mRNA的表达,Par-4蛋白的表达,Cas-pase-3P20活性片段,Caspase-3活性均有所增加。更多还原

【Abstract】 AIM:To explore the effects of H2O2 on the expression of gene par-4 and caspase-3 in PC12 cells,so as to provide experimental evidence for the mechanism of the incidence of neuronal apoptosis after Alzheimer’s disease(AD). METHODS:After the PC12 cells were treated with H2O2[0-400 nmol/L] for 8 hours, or treated with 200 nmol/L H2O2 from 0 to 8 hours,changes of mRNA expressions of par-4,caspase-3 were detected by RT-PCR,protein expression of Par-4 and the activity fragment of Caspase-3P20 were detected by Western-blot,Caspase-3 relative activities(A405nm) was determined by colorimetric method. RESULTS:The mRNA expression of par-4,caspase-3 gene and the protein expression of Par-4 and activity fragment of caspase-3 P20 were up-regulated with the increase of H2O2 concentration[0-400 nmol/L].After the PC12 cells were treated with 200 nmol/L H2O2 from 0 to 8 hours,the par-4 mRNA expression and Par-4 protein expression were increased with time from 2 to 8 hours;and the caspase-3 mRNA expression and Caspase-3 P20 activity fragment were increased with time 4 to 8 hours,respectively.The Caspase-3 relative activities(A405nm) was increased with the concentration of H2O2 from 50-400 nmol/L(χ2=4.8,P< 0.05),and reached its peak when the concentration was 400 nmol/L. CONCLUSION:Along with the dose-dependent increase of H2O2 and the increase of time at the same dose of H2O2 when dealing with PC12 cells,the mRNA expression of par-4 and caspase-3 gene,the protein expression of Par-4 and the caspase-3 P20 activity fragment and Caspase-3 relative activity are all increased.更多还原