【作者】 艾三喜；

【导师】 唐北沙；

【作者基本信息】 中南大学 ， 神经病学， 2012， 博士

【摘要】 目的研究散发性帕金森病(PD)患者外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)中SNCA基因的甲基化水平；研究SNCA甲基化水平、Repl多态及SNCA mRNA表达水平之间的相关性。方法密度梯度离心分离65例PD患者和63例正常人的PBMCs;重亚硫酸盐测序法检测SNCA基因内含子1区的甲基化水平；实时荧光定量PCR (real-time fluorescent quantitative PCR, RT-PCR)法检测SNCA mRNA的表达水平；直接测序法检测SNCA启动子区Rep1多态结果与正常对照组相比,PD患者PBMCs中SNCA基因内含子1区甲基化水平降低(P=0.024); Rep1等位基因267bp与SNCA内含子1区的高甲基化相关,Rep1等位基因271bp与SNCA内含子1区的低甲基化相关。271bp/271bp基因型较267bp/267bp和269bp/269bp基因型甲基化水平低。SNCA mRNA表达水平在PD组和对照组之间无显著统计学差异(P=0.107), SNCA mRNA表达水平与SNCA内含子1甲基化水平及Rep1多态无明显相关性。结论PD患者PBMCs中SNCA基因存在低甲基化。Rep1等位基因及基因型与SNCA内含子1区的甲基化水平相关。更多还原

【Abstract】 Objective:SNCA is a pathogenic gene identified in rare familial PD, and over-expression of SNCA was suggested in the pathogenesis of familial PD as well as sporadic PD. Allele length variability in Rep1of SNCA modulates susceptibility to sporadic PD and affects SNCA expression levels in both brains and peripheral blood mononuclear cells (PBMCs) of patients with sporadic PD. DNA methylation in SNCA intron-1is also involved in the regulation of SNCA expression and hypomethylation of this region was observed in postmortem brains of patients with sporadic PD. In this study, we studied the methylation status of SNCA intron-1, SNCA mRNA levels and Rep1genotypes in PBMCs of sporadic PD patients and controls and explored the relationship between DNA methylation, mRNA expression and Rep1genotypes.Methods:Peripheral blood mononuclear cells (PBMCs) from65sporadic PD patients and63healthy controls were isolated by density gradient centrifugation; DNA methylation level of SNCA intron-1was determined by bisulfite sequencing; mRNA level of SNCA was meatured by real-time quantitative PCR; Rep1polymorphism was examined by direct sequencing.Results:Hypomethylation of SNCA intron-1was detected in PBMCs of PD patients (p=0.024). And DNA methylation levels were associated with Rep1alleles and genotypes. The longest allele (271bp) was associated with lower level of methylation, while the shortest allele (267bp) was associated with higher level of methylation. Genotype 271bp/271bp showed significantly lower methylation levels than genotype267bp/267bp and269bp/269bp (p=0.014and0.043respectively). No significant difference of SNCA mRNA level was observed between PD and controls (p=0.107), and no correlation was observed between SNCA mRNA level and methylation level of SNCA intron-1or Rep1polymorphism.更多还原