【作者】 冷雪峰；

【导师】 李学英；

【作者基本信息】 遵义医学院 ， 生物化学与分子生物学， 2009， 硕士

【摘要】 目的:进一步研究遵义地区人群激素敏感脂肪酶(HSL)基因第4外显子多态性与2型糖尿病的关系,为上述疾病的发病机理和临床治疗补充相关理论依据。方法:采用两种实验设计,收集357例2型糖尿病患者和268例正常人血凝块各3毫升,碱裂解法提取血液细胞DNA;运用聚合酶链反应(PCR)扩增hsl第4外显子;结合单链构象多态性(SSCP)技术与RFLP技术对hsl第4外显子多态性进行进一步检测;实验结果进行统计学分析。结果:1.进一步验证了hsl第4外显子在遵义地区人群中具有多态分布。2.所选样本基因型频率经Hardy-Weinburg遗传平衡定律检验达到平衡,具有群体代表性。3.hsl外显子4-4A/G位点,所有样本:T2DM组检出GG基因型者51人,基因型频率为11.4%;AG基因型者157人,基因型频率为42.7%;AA基因型者149人,基因型频率为45.9%;对照组中,GG基因型者检出16人,基因型频率5.9%;AG基因型检出119人,基因型频率为44.3%;AA基因型检出133人,基因型频率为49.6%。随机病例-对照样本:T2DM组检出GG基因型者41人,基因型频率为13.5%; AG基因型者136人,基因型频率为44.2%;AA基因型者130人,基因型频率为42.3%;对照组中,GG基因型者检出10人,基因型频率4.7%;AG基因型检出98人,基因型频率为44.9%;AA基因型检出110人,基因型频率为50.4%。两组间三种基因型和两种等位基因的分布均存在显著性差异(P＜0.05);且GG基因型者占总人数的比列两组间比较差异显著(χ~2=9.027,P＜0.05和χ~2=8.46,P＜0.005); T2DM组人群GG基因型比例较高。4.在病例和家系对照组中,GG基因型者检出率分别为10人和6人,AG基因型者检出均为21人,AA基因型者检出率分别为19人和23人。结果无统计学差异(χ~2=1.381, P＞0.05)。5.将具有高脂血症者基因型和血脂正常组比较,经χ~2检验,两组基因型频率(χ~2=9.188, P＜0.05)和等位基因频率(χ~2=4.996,P＜0.05)差异显著,说明两组基因型和等位基因分布不同;且GG基因型人数占总人数的比例在高脂血症者和血脂正常两组间比较差异显著,高脂血症者组人群GG基因型比例较高(χ~2=7.129,P＜0.05)。6.将腰臀比异常与腰臀比正常组基因型比较,经χ~2检验,两组基因型频率(χ~2=8.886,P＜0.05)和等位基因频率(χ~2=4.164,P＜0.05)差异显著,说明两组基因型和等位基因分布不同;且GG基因型人数占总人数的比例在腰臀比异常与腰臀比正常两组间比较差异显著,腰臀比异常组人群GG基因型比例较高(χ~2=7.523,P＜0.05)。结论:1.遵义地区汉族人群hsl外显子4具有多态性分布。2.GG基因型是遵义地区汉族人群T2DM的遗传易感基因型。3.GG基因型和携带有等位基因G者可能是高脂血症、腹部脂肪堆积的一个危险因素。更多还原

【Abstract】 Objective: To investigate the relationship between the polymorphism hsl exon4 and type 2 diabetes mellitus in Han population of Zun Yi District. Our arms is to supplement the evidence for the pathogenesis of T2DM.Methods: Samples including in 268 health persons and 368 type 2 diabetes were chosen, all of above are Han population of Zun Yi District. Clinical data were collected from hospital. Blood clot was collected to detect hsl .DNA was extracted from nucleated cell with alkaline. Design special primer to amplify hsl exon4 by PCR. Then the product of PCR was used to detect polymorphisms by SSCP and RFLP. SPSS16.0 was used to analyze.Results:1.Multi-state distributions were found in the exon4 of the hsl in the Zun Yi District infurther.2.Hardy-Weinberg inherit equilibrium laws examination result showed that each genotypefrequency attains heredity balance and have a community representative.3.There was a statistically significant difference in the genotype distribution of -4 A/G polymorphisms of hsl and allele variations among the all subjects and random - control group. The frequencies of G allele and GG genotype for T2DM patient were significantly higher than those of controls(GG:11.4%vs5.1%; G:χ~2=15.47,P＜0.05). Among all subject, the T2DM group genotype GG was 11.4%, AG was 42.7% and AA was 45.9% .Comparatively, among the control group, The genotype GG was 5.1%, AG was 44.3% and AA was 49.6%. Random - control group: the T2DM group genotype GG was 13.5%, AG was 44.2% and AA was 42.3%. Comparatively, among the control group, the genotype GG was 4.7%, AG was 44.9% and AA was 50.4%.4.Among case-family group, the genotype GG was 20.0% and 12.0% respectively. The genotype AG were all 42.0%, AA was 38.0% and 46.0% respectively. Genotype distribution was no significant difference among the various groups (P＞0.05).5.In stratified analysis on the basis of blood-fat, the frequencies of -4 A/G genotype and G allele for hyperlipoidemia group were significantly higher than those of controls(χ~2=9.188, P＜0.05,χ~2=4.996, P＜0.05).6.In stratified analysis on the basis of WHR, the frequencies of -4 A/G genotype and G allele for abnormality WHR group were significantly higher than those of controls(χ~2=8.886,P＜0.05;χ~2=4.164,P＜0.05).Conclusion:1.Multi-state distributions was indicated in the exon4 of the hsl in Zun Yi District.2.The genotype GG is the major of genotype for hereditary susceptibility of T2DM in ZunYi District.3.The genotype GG and those carrying the G allele could be a risk factor for hyperlipoidemia and abdominal fat accumulation.更多还原