摘要
目的:探讨尿液检测局灶节段性肾小球硬化足细胞损伤与其他足细胞病之间的特点和差异。方法:入选原发性局灶节段性肾小球硬化(FSGS)患者54例,膜性肾病(MN)23例及微小病变(MCD)12例,正常对照20例。免疫荧光法计数尿足细胞,荧光实时定量PCR法定量尿沉渣足细胞相关分子nephrin、podocin、synaptopodin mRNA的表达水平,Western印迹法检测尿液WilmsTumor1(WT1)蛋白水平,免疫荧光法检测肾脏组织podocalyxin的表达及分布。结果:(1)FSGS组、MN组、MCD组和对照组尿足细胞阳性率分别是63%、34.8%、33.3%和0,FSGS组与其余各组相比差异均有统计学意义(P<0.05)。FSGS组足细胞脱落数目显著高于MCD组、MN组和对照组(P<0.05),伴足细胞尿FSGS患者与不伴足细胞尿FS-GS患者相比,24h尿蛋白和血清白蛋白(Alb)差异均有统计学意义(P<0.05)。(2)FSGS组尿沉渣足细胞nephrin mRNA表达水平显著高于MCD和MN组(P<0.05);FSGS组尿沉渣足细胞podocin mRNA表达显著高于MCD组(P<0.05),与MN组相比有升高趋势但差异无统计学意义;尿沉渣足细胞synaptopodin mRNA表达各组间差异无统计学意义。尿沉渣足细胞nephrin、podocin、synaptopodin mRNA的表达与24h蛋白尿无相关性。(3)FSGS组尿WT1蛋白量显著高于MCD和MN组。部分足细胞阴性患者尿液检测到WT1分子。(4)FSGS患者肾组织podocalyxin较对照组、MCD和MN有明显的节段缺失。结论:局灶节段性肾小球硬化病患者足细胞损伤严重,尿足细胞与FSGS疾病活动相关。尿沉渣足细胞nephrin mRNA表达可以把FSGS与MCD和MN区分开来,尿WT1蛋白可能是足细胞早期损伤指标。
Objective： To detect podocyte injury in the urine of focal segmental glomeruloscleros （FSGS） and other podocyte diseases. Methods： Urinary samples were taken from 54 patients with FSGS, 23 with membranous nephropathy（MN） and 12 with minimal- change disease（MCD）and samples from 20 healthy volunteers were served as controls. Freshly voided urine samples were examined by immunofluorescence labeling, using monoclonal antibody against human podocalyxin. Urinary sediment podocyte associated moculars mRNA expression were quantified by Real- time PCR, podccyte associated molecule-Wilms Tumor 1 （WT1）protein expression were quantified by we.stern blot. The expression of podocyte associated molecule - podocalyxin in glomeruli were determined by immunofluorescence. Results：NO podocyturia was found in healthy controls. The negative rate of patients with FSGS was 63 %, that of patients with MN 34.8 % and that of patients with MCD 33.3 %. Patients with FSGS had significantly higher levels of urinary podocytes than those with MCD, MN and controls（P 〈 0.05）. The FSGS patients with podocyturia showed significant difference in 24 h proteinuria, plasm albumin level （P 〈 0.05）. The expression of nephrin mRNA in urinary sediment of FSGS was higher than those of MCD and MN （P 〈 0.05）. There were significant differences in urinary sediment podocin mRNA expression between patients with FSGS and MCD, The expression of synaptopodin mRNA in urinary sediment was not significant differences among these groups. The expression of podocin, nephrin and synaptopodin mRNA in Urinary sediment were not correlated with proteinuria. Urinary WT1 was significantly higher in FSGS than those in the MN and MCD patients. WT1 were also detected in some patients without podocyturia. Depletion of podocalyxin expression on glomerulus of FSGS patients was significant compared with that in the controls, MN and MCD patients. Conclusion： The podocytes damage of FSGS was severer compared with those of MN and MCD. Urinary podocyte of FSGS patients also correlated with disease activity. MCD can be distinguished from FSGS and MN by the expression of nephrin mRNA in urinary sediment. Urinary WT1 protein might be a useful biomarker to detect early podocyte injury.
出处
《中国中西医结合肾病杂志》
2009年第6期489-493,I0002,共6页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
上海市卫生局重点学科基金资助项目(No.05Ⅲ001)
上海市重点学科建设基金资助项目(No.T0201)
上海市领军人才基金
上海市科委重点项目(No.07JC14037)
