【摘要】 目的 研究我国南方汉族IgA 肾病患者FcαR1基因5′端调控及UTR区SNPs及其单倍型与我国南方IgA肾病患者预后的关系。方法 采集266例肾活检证实的IgA肾病患者血样,提取基因组DNA。用PCR产物直接测序法鉴定基因型,每3~6个月查尿蛋白、血肌酐及其他指标随访,以血肌酐浓度比基础值升高1倍以上或死亡作为随访终点。采用单因素相关分析及Logistic多元回归分析各位点多态性及其单倍型与肾功能恶化及预后的关系。结果 (1)该区域单倍型对及-27、56位点多态性与肾功能进展显著相关;(2) -27位点基因型TC与TT比较差异显著(P=0.003); 56位点基因型CC与TT比较,P=0.011;单倍型对TCC/CTC与TTT/TTT比较,P=0.000。结论 FcαR1基因第一外显子C56T多态位点基因型与我国南方IgAN患者预后相关。 【关键词】 IgA肾病; FcαR1基因;多态性; 预后;相关分析 Abstract:Objective To investigate the relationship between the prognoses and the polymorphisms of FcαR1 gene in Chinese Han IgA nephropathy (IgAN) patients in the South China.Methods 266 blood samples from renal?biopsies that had been proven IgA nephropathy were collected,the genotypes were determined by PCR product sequencing.All patients were followed up every 3 to 6 months by examining urinary protein, Scr, and other indicators.Follow?up was terminated when Scr value increased by one fold than the normal or in the case of the death of the patients.Follow?up lasted for more than six months.The relationship between the renal function and the polymorphisms of FcαR1 gene were evaluated by single factor analysis and multiple regression analysis with SPSS 10.0.Results (1)Haplotypes and -27, 56 loci were significantly correlated with the aggravation of renal function.(2)Genotype TC vs.TT in -27 locus, CC vs.TT in 56 locus, haplotypical TCC/CTC vs.TTT/TTT were different significantly on the aggravation of renal function in Chinese Han IgAN patients in the South China.Conclusion SNPs and it′s haplotypes of FcαR1 gene in 5′regulatory region and UTR region were associated with the prognoses of Chinese Han IgAN patients in the South China. Key words:IgAN; FcαR1 gene; polymorphism; prognosis; correlation analysis 资料显示,约15 %~40 %IgA 肾病( immu?noglobulin A nephropathy, IgAN) 患者在确诊后的20年内逐渐进展到终末期肾衰(end stage renal failure,ESRF)。最近的一项研究表明,影响IgAN 预后的主要指标依次为:伴有高血压的男性患者;每天大于1 g的蛋白尿;肾功能损伤(肾小球滤过率小于60 mL/min);严重的肾小球病理损伤(如严重系膜增殖、肾小球硬化、新月体形成和血管硬化等)[1]。近年来越来越多的研究表明, 遗传因素不但影响对该病的易感性, 且影响疾病的进程。已有研究发现包括ACE基因、ET?1及其受体基因多态位点与IgAN的进展相关[2-4]。 1.1 研究对象 1.2 方法 1.2.1 全血基因组DNA提取 按照QIAamp DNA Blood Maxi Kit试剂盒说明书操作。采用紫外分光光度法测定提取DNA的浓度与纯度。 1.2.2 PCR 扩增目的区域 根据FcαR1基因的核苷酸序(GenBank 序列号:X59875),设计扩增引物:上游引物5′?GCC CGT CTA TTT GGA TGT ?3′;下游引物5′?GGA GGG TGG TCT GTT TGG?3′。 反应体系为:10×PCR缓冲液(无镁) 2.5 μL、50 mmol/L MgCl2 1.5 μL、2 mmol/L dNTP混合物2.5 μL、10 μmol/L上、下游引物各1 μL、Taq DNA多聚酶(1 U/μL) 1.1 μL、DNA模板1 μL、灭菌去离子水加至25 μL;反应条件:94 ℃预变性3 min,94 ℃变性30 s,55 ℃退火30 s,72 ℃延伸45 s,共30个循环,最后72 ℃延伸5 min。 1.2.3 FcαR1 5′端调控及UTR区SNPs多态性的鉴定、分型 PCR扩增含有FcαR1 5′端调控及UTR区单核苷酸多态性区域,反应体系为:10×PCR缓冲液(无镁) 2.5 μL、50 mmol/L MgCl2 1.5 μL、2 mmol/L dNTP混合物2.5 μL、10 μmol/L |