【摘要】 目的:检测不同病理类型散发性难治性肾病综合征(NS)患者NPHS2基因突变。方法:研究对象为1997年至2004年我院收治的19例散发性难治性NS患者,年龄4～48岁,男性15例,女性4例,其中激素不敏感患者7例,包括局灶节段性肾小球硬化(FSGS)5例,微小病变(MCD)1例,系膜增生性肾小球肾炎(MsPGN)1例;激素依赖患者12例,包括MCD6例,FSGS4例,未行肾穿刺2例。外周血基因组DNAPCR扩增后直接测序。间接免疫荧光法检测肾组织中podocin的表达。结果:①发现1例NPHS2基因431C>A的杂合突变,造成144位氨基酸由脯氨酸转变为组氨酸,查阅文献和基因库,未发现相同突变报道。该患者为激素依赖性NS,病理类型为FSGS。正常对照102条染色体中未发现同样突变。②发现11个SNP位点,其中3个位于外显子区,4个位于内含子区,1个位于启动子区,1个位于5’-UTR区,2个位于3’-UTR区。③该患者肾组织中podocin表达明显减弱。结论:在1例激素依赖NS患者中发现1个新的NPHS2基因突变,且肾组织中podocin蛋白表达明显减弱。推测该例患者发病和激素治疗反应差可能与NPHS2突变有关。更多还原

【Abstract】 Objective: Studies of genetic background for refractory nephrotic syndrome have associated genes encoding proteins in slit membrane of podocyte foot processes with differe nt types of familial and sporadic glomerulopathies. Mutations of NPHS2 gene, e ncoding podocin, have been demonstrated by various studies to be responsible f or sporadic steroid-resistant nephrotic syndrome. In this study, we studied NP HS2 mutation in Chinese patients with sporadic refractory nephrotic syndrome, a nd discovered a noval mutation of NPHS2 (431C>A) in a Chinese with focal and seg mental glomerulosclerosis(FSGS). Methodology:Genomic DNA extracted from pheripheral blood cells was amplified by PCR, and mut ations of NPHS2 was analyzed by direct DNA sequencing. Nineteen patients with sp oradic refractory nephritic syndrome were included in this study, with 15 males and 4 females, aged between 4 and 48 years. All of them were hospitalized betwee n 1997 and 2004. 17 of the 19 patients underwent a renal biopsy for pathologic c lassification. Among them, 7 were grouped as steroid-insensitive (5 classified as FSGS, 1 as minimal change disease, and 1 as mesangial proliferative glomerulo nephritis), and 12 grouped as steroid-dependent (6 classified as minimal change disease, 4 as FSGS, 2 unclassified). 51 healthy subject were included in the c ontrol group. Results:A novel NPHS2 heterozygous mutation ( 431C>A), causing the 144th amino acid prol ine to be substituted by histidine, was detected in a patient with FSGS grouped as steroid-dependent. No such mutation was detected in any of the control group . Eleven SNPs were detected in this group of patients, including 3 located in ex on, 4 in inexon, 1 in the promoter region, 1 in 5-UTR, and 2 in 3-UTR. Imm unofluescence staining of biopsy slides with monoantibody to podocin revealed th at a significant decreament of expression of podocin in glomerular region in all the patients. Conclusion:In this study, we reported a novel NPHS2 heterozygous mutation in a FSGS patient who is steroid dependent. And, we found significant decrement of podocin expres sion in all patients with sporadic refractory nephrotic syndrome.更多还原