高同型半胱氨酸血症对膜性肾病大鼠肾组织Nephrin WT1表达的影响

The Effects of Hyperhomocysteinemia on the Expression of Nephrin and WT1 in Membranous Nephropathy Rats Renal Tissue

目的:观察高同型半胱氨酸血症对膜性肾病(MN)大鼠肾组织Nephrin、WT1表达的影响,探讨MN足细胞损伤凋亡的可能发生机制。方法:将30只SD雄性大鼠随机分为正常组和模型组,正常组不给予任何处理,模型组尾静脉注射16mg/kg阳离子化牛血清白蛋白(c-BSA)复制MN大鼠模型,于正式免疫第1,2,3,4周末检测24h尿蛋白(24h UTP)水平,并于第4周末检测大鼠血浆尿素(Ure)、血肌酐(Cre)及同型半胱氨酸(Hcy)水平。同时取材鉴定大鼠肾组织成模情况,免疫组化检测大鼠肾组织中Nephrin、WT1的表达,结果:模型组大鼠造模第4周成模稳定,病理表现典型。随造模时间延长,模型组24h UTP呈逐渐增加趋势,与前一周末比较差异均有统计学意义(P<0.01),2、3、4周末24h UTP较正常组有明显增加(P<0.01)。Nephrin在模型组肾组织中表达较正常组显著减少(P<0.01),WT1的表达与正常组比较无显著差异。HCY与肾组织Nephrin的表达呈明显负相关,与第4周末24h UTP呈明显正相关,与血浆尿素、血肌酐、肾组织WT1无明显相关关系。结论:MN中高Hcy血症可能通过下调Nephrin的表达参与了足细胞的损伤凋亡过程,积极监测Hcy水平及控制高Hcy血症有利于防止MN的进展。

Objective： To observe the effects of hyperhomocysteinemia（ HHcy） on the expression of Nephrin and WT1 in renal tissues of MN rats,and to investigate the possible mechanism of MN podocytes apoptosis. Method： 30 SD male rats were randomly divided into normal group and model group,normal group without any treatment,Model group was duplicated by 16 mg / kg C-BSA tail vein injection on the basis of modified Border method. 24 h urinary protein levels（ 24 h UTP） were detected at the first,second,third and fourth weekend in formal immunity. Plasma urea（ Ure）,serum creatinine（ Cre） and homocysteine（ Hcy）levels were detected at the fourth weekend. The expression of WT1 and Nephrin in rats renal tissues were detected by immunohistochemistry,and appraisal the MN model of rat kidney tissue. Result： The model is stable and the pathology is typical at the fourth weekend. 24 h UTP gradually increased with the increase of the model time,and increased significantly compared to normal group at the third weekend（ P〈0. 01）. Nephrin expression in the model group was significantly decreased compared with the normal group（ P〈0. 01）,the expression of WT1 was not significantly different from the normal group. The Hcy levels were significant negatively correlated with the nephrin expression,positively correlated with 24 h UTP in fourth weeks,no significant correlation with Ure,Cre and expression of WT1. Conclusion： HHcy may down-regulate the expression of nephrin to participate in the apoptosis of the podocyte,actively monitor the levels of Hcy and control the HHcy to prevent the development of MN.