【作者】 胡之琳；

【导师】 金美玲； 揭志军；

【作者基本信息】 复旦大学 ， 呼吸内科， 2008， 硕士

【摘要】 研究背景:哮喘是一种多基因遗传病,由基因和环境因素共同作用所决定。目前已发现几百个相关基因,ADAM33基因是目前研究较多的与哮喘相关的易感基因。研究表明ADAM33基因与支气管哮喘和气道高反应性相关,但其SNPs在不同种族人群中与支气管哮喘及其相关表型的联系不尽相同,而ADAM33基因S1、S2位点的SNPs与华东地区汉族人群支气管哮喘的发生,及其与肺功能的关系尚未有相关研究。研究目的:本研究采用病例-对照的研究方法,以中国华东地区汉族人群正常个体和散发支气管哮喘患者为研究对象,在ADAM33基因内部选择S1、S2多态位点,采用统计学方法比较病例组与对照组间、病例组不同肺功能患者间基因型、等位基因、单体型频率的差异,以探讨ADAM33基因多态性与中国华东地区汉族人群支气管哮喘是否具有相关性,及其多态性是否与肺功能相关。研究方法:本研究收集了散发的支气管哮喘患者147例,健康对照组73例,均抽取外周静脉全血,进行S1、S2位点基因分型。随访哮喘组,并进行分级治疗,直至肺功能不能进一步改善为止,取随访期间最佳FEV₁。定义哮喘组中FEV₁实测值/预计值≥80%为肺功能正常组,＜80%为肺功能异常组;定义FEV₁实测值/预计值＜50%为肺功能严重受损组。将受试对象分为哮喘组/健康对照组、肺功能正常组朋市功能异常组、肺功能严重受损组/健康对照组。比较各组间性别、年龄的可比性。计算各组S1、S2位点基因型频率及等位基因频率,进行Hardy-Weinberge检验,比较组间基因型、等位基因频率差异。对S1、S2位点进行连锁不平衡分析,并分析组间单体型频率差异。比较哮喘组不同基因型患者间FEV₁差异。比较华东地区汉族人群健康对照组S1、S2位点等位基因频率与不同种族人群健康对照组的差异。定义P≤0.05时,判定差异（或者联系）有统计学意义。结果:各组间性别、年龄均匹配,具有可比性。S1位点不同基因型及等位基因在哮喘组及健康对照组频率分布:哮喘组基因型A/G 12.2%,G/G 87.7%,等位基因A 6.1%,G 93.9%;健康对照组基因型A/G8.2%,G/G 91.8%,等位基因A4.1%,G 95.9%。S2位点不同基因型及等位基因在哮喘组及健康对照组频率分布:哮喘组基因型C/C 4.1%,C/G 33.3%,G/G62.6%,等位基因C 20.7%,G 79.3%;健康对照组基因型C/C 4.1%,C/G 35.6%,G/G 60.3%,等位基因C 21.9%,G 78.1%。两组基因型及等位基因频率相比较,均无统计学差异。S1位点不同基因型及等位基因在哮喘患者中肺功能正常组与肺功能异常组频率分布:肺功能正常组基因型A/G 8.9%,G/G 91.1%,等位基因A 4.4%,G95.6%;肺功能异常组基因型A/G 16.2%,G/G 83.8%,等位基因A 8.1%,G 91.1%。S2位点不同基因型及等位基因在肺功能正常组与肺功能异常组频率分布:肺功能正常组基因型C/C 3.8%,C/G 34.2%,G/G 62.0%,等位基因C 20.9%,G 79.1%;肺功能异常组基因型C/C 4.4%,C/G 32.4%,G/G 63.2%,等位基因C 20.6%,G79.4%。两组基因型及等位基因频率相比较,均无统计学差异。S1位点不同基因型及等位基因在肺功能严重受损组与健康对照组频率分布:肺功能严重受损组基因型A/G 8.3%,G/G 91.7%,等位基因A 4.1%,G 95.9%;健康对照组基因型A/G 8.2%,G/G 91.8%,等位基因A 4.2%,G 95.8%。S2位点不同基因型及等位基因在肺功能严重受损组与健康对照组频率分布:肺功能严重受损组C/G 29.2%,G/G 70.8%,等位基因C 21.9%,G 78.1%;健康对照组基因型C/C 4.1%,C/G 35.6%,G/G 60.3%,等位基因C 14.6%,G 85.4%。各组均符合Hardy-Weinberg定律,且组间基因型频率、等位基因频率无统计学差异。两组基因型及等位基因频率相比较,均无统计学差异。S1、S2位点间为弱连锁不平衡,各组间单体型频率无统计学差异。S1位点基因型A/G的FEV₁为实测值/预计值78.0±22.1%,G/G为77.4±22.7%;S2位点基因型C/C的FEV₁为实测值/预计值78.7±23.5%,C/G为78.3±21.3%,G/G为77.0±23.3%;组间比较无统计学差异。S1位点英国高加索人、美国高加索人、荷兰高加索人、美国黑人、美国白人、德国人等位基因频率与华东地区汉族人群相比,有统计学差异;美国西班牙人、墨西哥人、波多黎各人、韩国人等位基因频率与华东汉族人群比较,无统计学差异。S2位点美国黑人、美国白人、德国人等位基因频率与华东汉族人群比较,有统计学差异;英国高加索人、荷兰高加索人、美国西班牙人、日本人等位基因频率与华东汉族人群比较,无统计学差异。结论:ADAM33基因S1、S2位点SNPs与中国华东地区汉族人群支气管哮喘的发生无显著相关性,与肺功能指标FEV₁实测值/预计值无显著相关性。更多还原

【Abstract】 Background:Asthma,a polygenic hereditary disease,is induced by both hereditary factor and environmental factors.Recently,hundreds of genes have been associated with asthma, and ADAM33 gene is one of the hot spots,it is associated with asthma and bronchial hyperresponsiveness.The SNPs of ADAM33 gene,however,differ in their associations with the development of asthma and the related phenotypes in populations.To date,association of polymorphisms in locus SI&S2 with the development of asthma and pulmonary function has not been studied in Chinese Han population.Objective:Unrelated asthmatics and healthy controls of Huadong Chinese Han population were included in this case-control study.To investigate whether polymorphisms of locus SI&S2 in ADAM33 gene are associated with asthma and pulmonary function, the genotype frequencies and allele frequencies of different groups were analyzed.Methods:The study included 147 unrelated asthma patients and 73 healthy controls. Peripheral venous blood samples were taken,and the genotypes of locus S1&S2 were tested.We followed up the asthmatics,that were taken graded treatment,till there’s no further improvement of their pulmonary function,and the best FEV₁ results in this period were recorded.The asthmatics were classified into two groups: normal-pulmonary-function asthmatics（FEV₁ actual value/ predicted value≥80%） and decline-pulmonary-function asthmatics（FEV₁ actual value/ predicted value＜80%）.And the asthmatics,whose FEV₁ actual value/ predicted value＜50%, were grouped into sever asthmatic group.The subjects were compared in three different ways:asthmatics/healthy controls,normal-pulmonary-function asthmatics/ decline-pulmonary-function asthmatics,sever asthmatics/ healthy controls.We compared the sex and age of various groups,and evaluated each variation for deviations from Hardy-Weinberg equilibrium.The genotype distributions and allelic frequencies for studied polymorphisms for the various groups were statistically compared.The linkage disequilibrium was measured between locus S1 and locus S2, and frequencies of haplotypes were compared between groups.FEV₁ of asthmatics with different genotypes were compared.The allelic frequencies of healthy controls were compared between populations.Results were considered statistically significant when the probability of findings occurring by chance was less than 5%（P＜0.05）.Results:Sex and age of subjects in various groups were comparable.At locus S1,frequency of genotype A/G,G/G were 12.2%,87.7%,and frequency of allele A,G were 6.1%,93.9%in asthmatics,while frequency of genotype A/G,G/G were 8.2%,91.8%,and frequency of allele A,G were4.1%,95.9%in healthy controls. At locus S2,frequency of genotype C/C,C/G,G/G were 4.1%,33.3%,62.6%,and frequency of allele C,G were 20.7%,79.3%in asthmatics,while frequency of genotype C/C,C/G,G/G were 4.1%,35.6%,60.3%,and frequency of allele C,G were4.1%,95.9%in healthy controls.Frequency of genotype and allele were not statistically significant.At locus S1,frequency of genotype A/G,G/G were 8.9%,91.1%,and frequency of allele A,G were 4.4%,95.6%in normal-pulmonary-function asthmatics,while frequency of genotype A/G,G/G were 16.2%,83.8%,and frequency of allele A,G were8.1%,91.1%in decline-pulmonary-function asthmatics.At locus S2,frequency of genotype C/C,C/G,G/G were 3.8%,34.2%,62.0%,and frequency of allele C,G were 20.7%,79.3%in normal-pulmonary-function asthmatics,while frequency of genotype C/C,C/G,G/G were 4.1%,35.6%,60.3%,and frequency of allele C,G were 20.6%,79.4%in decline-pulmonary-function asthmatics.Frequency of genotype and allele were not statistically significant.At locus S1,frequency of genotype A/G,G/G were 8.3%,91.7%,and frequency of allele A,G were 4.1%,95.9%in sever asthmatics,while frequency of genotype A/G,G/G were 8.2%,91.8%,and frequency of allele A,G were 4.2%,95.8%in healthy controls.At locus S2,frequency of genotype C/G,G/G were 29.2%,70.8%, and frequency of allele C,G were 21.9%,78.1%in sever asthmatics,while frequency of genotype C/C,C/G,G/G were 4.1%,35.6%,60.3%,and frequency of allele C,G were4.1%,95.9%in healthy controls.Frequency of genotype and allele were not statistically significant.The measured LD was low between tested polymorphisms,and the frequencies of haplotypes were not statistically significant.At locus S1,FEV₁ actual value/predicted value of genotype A/G,G/G were 78.0±22.1%,77.4±22.7%.At locus S2,FEV₁ actual value/predicted value of genotype C/C,C/G,G/G were 78.7±23.5%,78.3±21.3%,77.0±23.3%.No significant difference was observed.At locus S1,compared to Huadong Chinese Han population,the allelic frequencies were statistically significant in UK Caucasian,US Caucasian,Dutch Caucasian,African American,US white and Germany,while the allelic frequencies were not statistically significant in US Hispanic,Mexican,Puerto Rican and Korean. At locus $2,compared to Huadong Chinese Han population,the allelic frequencies were statistically significant in African American,US white,Germany,while the allelic frequencies were not statistically significant in UK Caucasian,Dutch Caucasian,US Hispanic and JapaneseConclusion:The polymorphisms of locus S 1 &S2 showed no association with the development of asthma and pulmonary function in Huadong Chinese Han population.更多还原