论文部分内容阅读
目的:了解WT1基因突变在3例激素耐药肾病伴泌尿生殖器异常患儿的临床特点和致病作用。方法:采用间接免疫荧光及免疫组织化学的方法对2例患儿的肾组织行足细胞分子(nephrin,podocin,α-actinin4,WT1及CD2AP)表达;采用PCR及RT-PCR的方法检测WT1基因突变及+KTS/-KTS比例。结果:3例患儿发病年龄分别为6月、1岁及10岁,确诊年龄分别为7月、9岁及15岁。例1和例3为男性表型伴泌尿外生殖器异常,例2为女性表型;3例患儿染色体核型均为46,XY。临床均为激素耐药肾病,例1和例2肾脏病理为局灶节段性肾小球硬化(FSGS)。对2例肾活检患儿肾组织标本的分析显示足细胞分子表达均发生改变;例1WT1无表达,例2WT1在足细胞核内的分布与正常对照不同。WT1基因分析示,WT1基因序列中例1未发现突变,例2为IVS9+5G>A杂合突变,例3为WT1外显子91186G>A的杂合突变。结论:对于早发激素耐药肾病且病理为FSGS的女性患者或伴有泌尿生殖器异常的男性患者应行染色体核型和WT1基因分析。WT1突变引起足细胞分子表达发生改变,提示这些足细胞分子参与了蛋白尿的形成或发展。
Objective: To understand the clinical features and pathogenicity of WT1 gene mutation in 3 cases of steroid resistant nephropathy with genitourinary abnormalities. Methods: The expressions of nephrin, podocin, α-actinin4, WT1 and CD2AP were detected in 2 kidneys by indirect immunofluorescence and immunohistochemistry. The expression of WT1 gene was detected by PCR and RT-PCR Mutation and + KTS / -KTS ratio. Results: The onset ages of the three cases were 6 months, 1 year and 10 years old respectively, and the diagnosis ages were July, 9 and 15 years respectively. Cases 1 and 3 were male phenotypes with genitourinary abnormalities, case 2 was female phenotypes; and 3 children with chromosomal karyotypes were 46, XY. Clinical resistance to steroid resistant nephropathy, cases 1 and 2 renal pathology focal segmental glomerulosclerosis (FSGS). Analysis of 2 renal biopsy specimens revealed changes in the expression of podocytes. Example 1 WT1 was not expressed, and 2WT1 in podocyte nucleus was different from normal controls. WT1 gene analysis showed that no mutation was found in case 1 of WT1 gene sequence, case 2 was a heterozygous mutation of IVS9 + 5G> A, and case 3 was a heterozygous mutation of exon 91186G> A of WT1. CONCLUSIONS: Chromosomal karyotypes and WT1 gene analyzes should be performed on women with early-onset hormone resistant nephropathy and pathologically FSGS or in men with abnormal genitourinary status. WT1 mutations cause changes in the expression of podocytes, suggesting that these podocytes are involved in the formation or development of proteinuria.