散发性帕金森病患者外周血snca基因的甲基化研究

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1、中南大学博士学位论文散发性帕金森病患者外周血SNCA基因的甲基化研究姓名：艾三喜申请学位级别：博士专业：神经病学指导教师：唐北沙20120530博士学位论文中文摘要散发性帕金森病患者外周血SNCA基因的甲基化研究摘要目的研究散发性帕金森病(PD)患者外周血单个核细胞(peripheralbloodmononuclearcells，PBMCs)中SNCA基因的甲基化水平；研究SNCA甲基化水平、Repl多态及SNCArnRNA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P

2、CR(real．timefluorescentquantitativePCR，RT-PCR)法检测SNCAmRNA的表达水平；直接测序法检测SNCA启动子区Repl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=O．024)；Repl等位基因267bp与SNCA内含子l区的高甲基化相关，Repl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。删mRNA表达水平在PD组和对照组之间无显著统计学差异(P=O．107)，删mRNA表达水平与

3、SNCA内含子1甲基化水平及Repl多态无明显相关性。结论PD患者PBMCs中SNCA基因存在低甲基化。Repl等位基因及基因型与删内含子1区的甲基化水平相关。关键词帕金森病，甲基化，SNCA内含子1，Repl博士学位论文英文摘要ABSTRACTObjective：SNCAisapathogenicgeneidentifiedinrarefamilialPD，andover-expressionofSNCAwassuggestedinthepathogenesisoffamilialPDaswellassporadicPD．AllelelengthvariabilityinR

4、eplofSNCAmodulatessusceptibilitytosporadicPDandaffectsSNCAexpressionlevelsinbothbrainsandperipheralbloodmononuclearcells(PBMCs)ofpatientswithsporadicPD．DNAmethylationinSNCAintron一1isalsoinvolvedintheregulmionofSNCAexpressionandhypomethylationofthisregionwasobservedinpostmortembrainsofpatien

5、tswithsporadicPD．Inthisstudy,westudiedthemethylationstatusofSNCAintron一1，SNCAmRNAlevelsandReplgenotypesinPBMCsofsporadicPDpatiemsandcontrolsandexploredtherelationshipbetweenDNAmethylation，mRNAexpressionandRep1genotypes．Methods：Peripheralbloodmononuclearcells(PBMCs)from65sporadicPDpatientsan

6、d63healthycontrolswereisolatedbydensitygradientcentrifugation；DNAmethylationlevelofSNCAintron．1wasdeterminedbybisulfitesequencing；mRNAlevelofSNCAwasmeaturedbyreal—timequantitativePCR；Replpolymorphismwasexaminedbydirectsequencing．Results：HypomethylationofSNCAintron．1wasdetectedinPBMCsofPDpat

7、ientsp=0．024)．AndDNAmethylmionlevelswereassociatedwithRep1allelesandgenotypes．Thelongestallele(271bp)wasassociatedwithlowerlevelofmethylation，whiletheshortestallele(267bp)wasassociatedwithhigherlevelofmethylation．GenotypeII薄士学位论文英文摘簧271bp／271bpshowedsign