原发性局灶节段性肾小球硬化患者ACTN4基因变异和多态性  

ACTN4 gene variation and polymorphism in patients with primary focal segmental glomerulosclerosis

作　　者：戴胜川[1] 王朝晖[1] 潘晓霞[1] 陈晓农[1] 王伟铭[1] 任红[1] 冯旗[2] 陈楠[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾脏科,200025 [2]中国科学院上海生命科学院国家基因中心

出　　处：《中华肾脏病杂志》2008年第2期108-114,共7页Chinese Journal of Nephrology

基　　金：上海市卫生局重点学科基金（05111001）;上海市重点学科基金（T0201）;上海市领军人才基金;上海市科委重点项目（07JC14037）;上海市青年科技启明星培养计划（03QD14021）

摘　　要：目的了解ACTN4基因变异和多态性在原发性局灶节段性肾小球硬化（FSGS）发病中的作用。方法选取FSGS患者82例，另设70例健康人作为对照组。盐析法提取外周血基因组DNA，PCR扩增后测序，与基因数据库进行匹配，寻找可能致病变异位点。氯酚法提取患者父母头发DNA，间接免疫荧光法检测患者肾组织辅肌动蛋白4（α-actinin-4）表达水平。单核苷酸多态（SNP）位点经Hardy-Weinberg平衡检验后行基因频率、基因型和临床表型关联分析。结果发现l例患者单核苷酸变异184T〉A（Ser62Thr），1例5UTR变异1．34C〉T。对照组和患者父母未发现相同变异。1个疾病易感SNP位点484±87C〉G。变异者肾组织α-actinin-4表达水平分别较对照组和非变异FSGS组下降。变异基因型和野生基因型尿蛋白量（24h）的差异有统计学意义[（7．90±1．60）比（4.50±0．46）g／24h，P〈0．01]。此外，还发现6个新的变异和另1个SNP位点，但未引起氨基酸改变。结论原发性FSGS患者中存在ACTN4基因变异位点和疾病易感SNP位点。ACTN4基因变异在原发性FSGS发病中可能起重要作用。Objective To investigate the effects of variation and polymorphism of gene ACTN4 on primary focal segmental glomerulosclerosis （FSGS）. Methods Eighty-two patients with primary FSGS were enrolled in the study, including 43 males and 39 females, ranging from 12 to 76 years old. Among these patients, 55 were diagnosed as nephrotic syndrome （NS）. Seventy healthy volunteers were as control group. Genomic DNA was extracted from peripheral blood cells of primary FSGS patients and from hair of their parents. ACTN4 variation was analyzed by PCR and direct sequencing. Variation was matched with GenBank. Hair DNA of the parents with novel mutation was sequenced and alpha-actinin-4 expression in kidney tissue of the patients was examined by immunofluorescence. With single nucleotide polymorphism （SNP） loci, after Hardy- Weinherg equilibrium test, allele association and the frequencies of genotypes were analyzed, then genotypes and phenotypes were analyzed, including urine protein, serum albumin, blood pressure and serum creatinine. Results In this study, one heterozygous variation 184T〉A with aminoacid substitution （Ser 62Thr） and one 5＇UTR mutation 1-34C〉T were detected, and both patients were diagnosed as non-NS FSGS. The above variation were not found in the control group. Matched with genbank, 1-34C〉T and 184T〉A might affect the transcription and translation of ACTN4 gene as well as alpha-actinin-4. No above variation were found in their parents＇DNA and alpha-actinin-4 expression reduced significantly in patients＇ kidney tissue （P〈0.01）. Additional 17 variation or SNP were also screened, including 6 novel variation, 2 novel SNP and 9 genbank reported SNP, without am/no-acid substitution. Novel SNP 484 ＋87C 〉G was associated with FSGS （Hardy-Weinberg equilibrium test, P〉0.05; G frequence 0.085 vs 0.029, P〈0.05）, and there was significant difference in urinary protein excretion between wild genotype and mutated genotype （7.90±1.60 g/ 24 h vs 4.50±0.46 g/24 h, P〈0.01）.

关 键 词：肾小球硬化症 病灶性 变异(遗传学) 多态性 单核苷酸 ACTN4基因 

分 类 号：R692.6[医药卫生—泌尿科学] R512.62[医药卫生—外科学]