【作者】 王学静；

【导师】 景强；

【作者基本信息】 昆明医学院 ， 法医学， 2011， 硕士

【摘要】 研究目的酒精依赖(AD)是一种多因子控制的复杂疾病,而超过一半的饮酒风险度由遗传因素决定。在与酒精依赖相关的神经递质中,5-羟色胺(5-HT)系统受到广泛关注。迄今对5-羟色胺2A受体(HTR2A)和5-羟色胺转运体(5-HTT)基因多态与酒精依赖相关性的研究报道虽多,但结论的矛盾性和差异性仍然存在,并且还未见云南省汉族人群中的相关研究报道。为此,本研究选择云南汉族人群对HTR2A和5-HTT 2个基因7个多态位点进行检测,以探讨其与酒精依赖发生的关系。研究方法按照DSM-Ⅳ诊断标准选择2006-2010年五年来云南省精神病医院酒与药物依赖治疗科住院治疗的118例酒精依赖综合征患者作为研究对象,同时选取与上述病例组相匹配的218例健康体检者作为对照。应用PCR-RFLP和PCR测序方法对336个云南汉族HTR2A基因中的rs6311、rs6313、rs6314、rs2296972、rs4941570和rs4941571 6个SNPs和5-HTT基因中的5-HTTLPR共7个位点进行了基因分型,采用SPSS16.0、SHEsis和Arlequin 3.01对结果进行统计和分析。研究结果1、单个位点分析结果在云南汉族人群中,HTR2A基因rs6311(A-1438G)、rs6313(T102C)、rs2296972、rs4941570和rs4941571位点均与酒精依赖无关；5-HTT基因启动子区域5-HTTLPR多态性与酒精依赖存在相关性,酒精依赖患者组和正常对照组的5-HTTLPR的基因型分布存在显著性差异,LL和LS基因的携带者人群嗜酒发生率显著低于SS基因型人群。S和L等位基因频率在两组间无统计学差异,但其分布存在种族差异性。2、组合基因型分析结果5-HTTLPR SL/rs6311 AA、SL/rs631377 TT、SL/rs4941571 CC基因型组合使酒精依赖患病风险降低,5-HTTLPR SS/rs6311 GG、SS/rs4941570AA基因型组合使酒精依赖患病风险降升高。结果提示5-HTTLPR与HTR2A基因位点具有协同作用。2、单倍型分析结果HTR2A基因的5个SNPs (rs6311、s6313、rs2296972、rs4941570和rs4941571)在云南汉族中构建出11种主要单倍型。总体检验显示HTR2A基因酒精依赖组和正常对照组间的单倍型分布具有显著性差异；H6(GCTGC)单倍型可能使酒精依赖的患病风险升高；H8 (ATGGC)在可能使酒精依赖的患病风险降低。3、连锁不平衡分析云南汉族人群中,HTR2A基因的rs6311、rs6313、rs4941570及rs4941571之间存在着强连锁(D’>0.8)。研究结论在云南汉族人群中,HTR2A基因rs6311(A-1438G)、rs6313(T102C)、rs2296972、rs4941570和rs4941571位点均与酒精依赖无关；5-HTT基因启动子区域5-HTTLPR多态性与酒精依赖存在相关性,LL和LS基因型可能是降低酒精依赖发病的影响因子之一5-HTTLPR与HTR2A基因存在协同作用。H6 (GCTGC)单倍型可能使酒精依赖的患病风险升高；H8 (ATGGC)在可能使酒精依赖的患病风险降低。云南汉族人群中,HTR2A基因的rs6311、rs6313、rs4941570及rs4941571之间存在着强连锁(D’>0.8)。更多还原

【Abstract】 Objective:Alcohol dependence (AD)is a complex disorder which is influenced by many factors.Studies indicate that hereditary factor plays a dominating role in the process of AD pathogenesis.There have been many research reports regarding the association between HTR2A and 5-HTT gene polymorphisms and AD, but the results are inconsistent and conflicting and never performed in Yunnan Han population. The aim of present study is to study the relation between the six SNPs of human HTR2A and 5-HTT gene polymorphism with alcohol dependence,and to study the etiopathogenesis of AD.Methods:According to the DSM-IV diagnostic criteria, we selected 118 alcohol dependent patients of Mental Health Center of Yunnan Province and 218 normal people as control group in the period during 2006-2010.A total of 7 variations(six SNPs of the HTR2A gene and 5-HTTLPR of the 5-HTT gene) were genotyped in 336 Yunnan Han individuals using PCR-RFLP and DNA sequencing techniques. The results are made statistical and biological analyzing by using SPSS 16.0、SHEsis and Arlequin 3.01.Results:1、The result of single locus analysisThere was no significant association of rs6311(A-1438G)、rs6313(T102C)、rs2296972、rs4941570 and rs4941571 of HTR2A gene between alcohol dependent group and control group in Yunnan Han Population.However,5-HTTLPR polymorphism of the 5-HTT gene might be associated with alcohol dependent patients. Significant differences in genotype frequencies were present between case group and control group of 5-HTTLPR.The proportion of LL and LS genotype was significantly smaller in case group than that was in control group.No significant association was observed in allelic frequencies, which differed in the different ethnic groups.2、The analysis results of combined haplotypesWhen SL of 5-HTTLPR combined with AA of rs6311,or TT of rs6313,or CC of rs4941571,the sick risk would be decreased, When SS of 5-HTTLPR combined with GG of rs6311, or AA of rs4941570,the sick risk would be increased.The results suggested there is a synergistic gene-gene interaction between 5-HTTLPR and HTR2A gene in the pathogenesis of AD.3、The analysis results of haplotypes for 5 SNPsThe results of haplotype analysis for rs6311、rs6313、rs2296972、rs4941570 and rs4941571 of HTR2A gene revealed that there are 11 kinds of major haplotypes in Yunnan Han Population.The total test showed there were significant differences in the HTR2A gene polymorphisms between alcoholics and controls. H6 (GCTGC) might be a dangerous factor leading to alcohol dependence,H8 (ATGGC) might be significantly associated with increased risks of alcohol dependence.4、The analysis results of lingkage disequilibriumLingkage disequilibrium analysis showed that there were strong linkages between rs631、rs6313、rs4941570 and rs4941571 in Yunnan Han Population. (D’ >0.8).Conclusion:There was no significant association of rs6311(A-1438G) rs6313(T102C)、rs2296972、rs4941570 and rs4941571 of HTR2A gene between AD group and control group in Yunnan Han Population. However,5-HTTLPR polymorphism of 5-HTT gene may be associated with alcohol dependent patients, and the genotype LL or LS may be a genetic factor that responsible for decreasing susceptibility of AD in Yunnan Han population.There is a synergistic gene-gene interaction between 5-HTTLPR and HTR2A gene in the pathogenesis of AD.H6 (GCTGC) might be a dangerous factor leading to AD,H8 (ATGGC) might be significantly associated with increased risks of AD.There were strong linkages between rs6311、rs6313、rs4941570 and rs4941571 in Yunnan Han Population. (D >0.8).更多还原