【作者】 张丽丽；

【导师】 寇长贵；

【作者基本信息】 吉林大学 ， 流行病与卫生统计学， 2019， 硕士

【摘要】 代谢综合征（Metabolic Syndrome,MetS）是一组复杂的、慢性的代谢紊乱性疾病,主要特征表现为中心性肥胖、血脂代谢紊乱、血压异常升高、血糖异常升高,但目前对该病的病因及发病机制的研究尚不十分明确。全基因组关联研究（Genome-wide association study,GWAS）显示同源体BUD13（BUD13 homolog,BUD13）基因上的单核苷酸多态性（Single nucleotide polymorphisms,SNPs）位点可调节血脂水平,从而有可能增加代谢综合征的患病风险。但目前关于BUD13基因多态性位点报道的较少,而且BUD13基因-环境之间交互作用与代谢综合征的关联性也不完全明确。因此,本研究将从环境因素、遗传因素和基因-环境交互作用三个方面,阐明代谢综合征的病因,为代谢综合征的进一步研究提供理论支撑。目的:了解中国北方汉族人群代谢综合征与BUD13 SNPs、环境因素和BUD13SNPs-环境交互作用的关联,阐明BUD13 SNPs与BUD13 SNPs-环境交互作用在代谢综合征发生的潜在机制,为代谢综合征的防治提供理论基础。方法:本研究采用大样本病例-对照研究,共纳入3850名中国北方汉族人群作为研究对象,其中病例组1813人,对照组2037人,性别与年龄均匹配。利用HapMap数据库筛选BUD13位点,再从中选择功能性位点,分别为rs7118999和rs10488698。抽取纳入所有研究对象的外周血,通过MALDI-TOF-MS技术和PCR方法检测所有个体的基因型。拟合优度c²检验计算SNPs基因型频数分布是否符合遗传平衡定律（Hardy-Weinberg定律）;SNP Stats在线分析软件计算SNPs的最优遗传模型;最优遗传模型下,Quanto遗传分析软件计算SNPs与代谢综合征关联性的功效;Logistic回归计算不同遗传模型下SNPs与代谢综合征的遗传关联性;协方差计算SNPs与病例组中代谢综合征组分间的关联性;结合筛查到的环境因素,Logistic回归计算SNPs-环境交互作用与代谢综合征的关联性。结果:1.均衡性检验:本研究病例组和对照组平均年龄分别为49.5±9.7岁和49.5±9.4岁,两组人群的年龄分布无统计学差异（t=-0.062,P=0.950）。病例组男性903人（占49.8%）,女性910人（占50.2%）;对照组男性1024人（占50.3%）,女性1013人（占49.7%）,性别在两组人群的分布无统计学意义（c²=0.082,P=0.774）。年龄和性别在两组中的分布均均衡可比。2.Hardy-Weinberg定律检验:BUD13基因上多态性位点rs7118999和rs10488698基因型的频数分布在病例组和对照组均符合Hardy-Weinberg定律（P>0.05）,说明本研究的样本具有代表性,适合做遗传学分析。3.SNPs与代谢综合征遗传关联性:BUD13基因rs7118999和rs1048869最优遗传模型均为隐性(AIC_min分别为4985.0和5316.8),该模型下rs7118999和rs1048869位点的功效分别为75.2%和84.1%。不同遗传模型下,SNPs在病例组和对照组间分布均无统计学差异（P>0.05）。4.SNPs与代谢综合征组分的关联性:BUD13基因SNPs基因型与病例组代谢综合征组分之间比较发现:rs7118999位点与腰围存在统计学差异（F=4.236,P=0.015）,且多重检验显示,TT>CT（P=0.023）;但未发现该位点与其他代谢综合征组分存在统计学差异（P>0.05）。rs10488698位点与高密度脂蛋白胆固醇存在统计学差异（F=5.660,P=0.004）,且多重检验显示,TT>CT（P=0.007）;但未发现该位点与其他代谢综合征组分存在统计学差异（P>0.05）。5.基因-环境交互作用:食盐摄入量、吸烟、饮酒和体育活动的主效应与代谢综合征存在统计学差异（P<0.05）,rs7118999/rs10488698-食盐摄入量/吸烟/饮酒/体育活动交互作用与代谢综合征均无统计学差异（P>0.05）。结论:1.不同遗传模型下,BUD13基因多态性位点rs7118999和rs10488698与代谢综合征均不存在遗传关联性;最优遗传模型均为隐性,该模型下,rs7118999和rs10488698位点的功效分别为75.2%和84.1%。2.BUD13基因的多态性位点rs7118999和rs10488698影响代谢综合征组分。携带rs7118999 TT基因型患者的腰围较CT基因型患者大,携带rs10488698 TT基因型患者的高密度脂蛋白胆固醇水平高于携带CT基因型患者。3.rs7118999/rs10488698与筛选出的各个环境因素（食盐摄入量、吸烟、饮酒和体育活动）之间均不存在交互作用,即BUD13 SNPs-环境交互作用不影响代谢综合征的疾病易感性。更多还原

【Abstract】 Metabolic syndrome(MetS)is a cluster of metabolic abnormalities,including central obesity,raised triglyceride levels,low high-density lipoprotein cholesterol levels,raised blood pressure,and raised glucose levels,and its pathogenesis remains undefined.Genome-wide association study indicates that BUD13 gene of single nucleotide polymorphism(SNPs)may adjust lipid level to lead to increase the risk for MetS.But few SNPs associated with MetS have been reported,and it is unclear that the association between MetS and gene-environment interaction.Therefore,this study reveals the cause of MetS through environment,gene and gene-environment interaction,and elucidates the etiology of MetS to provide a solid theoretical basis.ObjectiveIn order to explore potential pathogenesis of MetS,this study assesses whether environmental factors,BUD13 gene of SNPs and BUD13 SNPs-environment interaction link to MetS in the northeast of Chinese Han population.MethodsThe case-control study recruited 3850 Han population in northeast of China,including 1813 patients with MetS and 2037 healthy individuals.The distribution of age and gender matched.By using HapMap two functional SNPs were chosen on BUD13 gene including rs7118999 and rs10488698.DNA was isolated from peripheral blood samples using a commercial DNA extraction kit.Genotyping was determined by MALDI-TOF-MS and PCR.Hardy-Weinberg disequilibrium was tested by goodness-of-fitsχ2 test with 1 degree of freedom.The online SNP Stats program evaluated best inheritance model.The genetic power was calculated using Quanto software in best inheritance model.Logistic regression analysis evaluated the association of the chosen SNPs with MetS by adjusted age and gender in different inheritance model.The association of the investigated SNP with individual components of MetS used univariate tests by adjusted age and gender.The interactions between environmental factors and BUD13 SNPs with MetS were assessed using logistic regression analysis adjusted age and gender.Results1.The equilibrium test of age and genderThe average age were 49.5±9.7 and 49.5±9.4 in case and control groups respectively.There were 903 male(49.9%)and 910 female(50.2%)among 1813patients with MetS.The number of male and female subjects were 1024 and 1013 in control group,accounting for 50.3%and 49.7%respectively.There were no significant difference in age(t=-0.062,P=0.950)and gender(c~2=0.082,P=0.774)between case and control groups.2.Hardy-Weinberg equilibrium testThe distributions of rs7118999 and rs10488698 conformed to Hardy-Weinberg equilibrium in both case and control group(all P>0.05),thus these samples were suitable of genetic analysis.3.Associations between SNPs and MetSWe found that the best inherence model for rs7118999 and rs1048869 were recessive model,and the genetic power of rs7118999 and rs1048869 were 75.2%and84.1%in recessive model.There were no significant association between SNPs and MetS in different heritance models after adjusted age and gender between case and control group(P>0.05).4.Associations between SNPs and MetS componentsRs7118999 was associated with waist circumference(WC)in case group(F=4.236,P=0.015)and the carrier of TT was significantly associated with higher level of WC than CT(P=0.023)after adjusted age and gender.However,rs10488698 linked to high density lipoprotein cholesterol(HDL-C)in case group(F=5.660,P=0.004)and rs10488698 of TT was significantly associated with higher level of HDL-C than CT(P=0.007).But,our results did not showed other association between the two SNPs with the rest of MetS components after adjusted age and gender in case group(P>0.05).5.Gene-Environment interactionThis study showed that excess salt intake,smoking,drinking and no physical exercise contributed to MetS,but there were no interaction between rs7118999/rs10488698 in recessive model and above environment factors with MetS using logistic regression model after adjusted age and gender(P>0.05),which indicated that BUD13SNPs–Environment interaction did not confer to MetS risk.Conclusion1.This study indicated that two novel mutations locis in BUD13 did not confer risk for MetS in the northeast of Chinese Han population and the best inherence model of rs7118999 and rs10488698 were recessive model.The genetic power of rs7118999 and rs10488698 were 75.2%and 84.1%in recessive model.2.This study demonstrated BUD13 gene of SNPs mutations(rs711899 and rs10488698)changed the level of metabolic complements.The carrier of TT in rs7118999 conferred to risk of MetS through increasing the level of WC.While the carrier of TT in rs10488698 might be a protective factor for MetS through improving high level of HDL-C.3.This study showed rs7118999/rs10488698-environment(Salt intake,smoking,drinking and physical exercise)interaction did not occurred in our individuals,which indicated these interaction might not be pathogenesis for metabolic syndrome.更多还原