nephrin及WT-1在嘌呤霉素大鼠肾病模型足细胞中的表达及意义

目的应用嘌呤霉素氨基核苷（PAN）模型，研究足细胞相关蛋白nephrin和肿瘤蛋白-1（WT-1）在足细胞中的表达及在肾小球硬化进展过程中的意义。方法单次尾静脉注射嘌呤霉素氨基核苷制作PAN模型，在4、14及20周处死大鼠，取其肾脏，切片，nephrin及WT-1免疫组化染色，半定量分析后进行统计学分析。结果对照组免疫组化染色nephrin沿肾小球毛细血管襻呈较为均匀的黄褐色粗颗粒样的线状分布，4周时与对照组比较PAN组nephrin略有分布不均，部分区域染色减弱或消失（均值为10．23±1．256和8．23±0．88，P〈0．05），14周时在有节段性硬化的区域nephrin无表达（均值为7．58±0．71和10．23±1．26，P〈0．05），而在20周时硬化的区域基本无表达（均值为7．88．±0．92和6．08±1．34，P〉0．05），与对照组比较均有明显减少。4周时WT-1在足细胞核表达，呈深棕黄色粗大颗粒，与对照组比较无统计学意义（均值为10．14±2．02和10．11±1．56，P〉0．05）；14周时PAN组WT-1表达较对照组有所减少（均值为9．02±1．73和6．8±1．89，P〈0．05）；20周时与对照组比较PAN组WT-1明显减少（均值为7．34±1．22和4．43±1．01，P〈0．05），足细胞数量随着时间的延长依次递减。结论Nephrin的表达在肾小球硬化过程中进行性减少，甚至消失；WT-1的表达随着肾小球硬化的进展逐渐减少。Nephrin和W-1表达的降低均反映足细胞的结构破坏和数量的减少，因此两者表达可反映足细胞损伤和肾小球硬化的关系。

Study on the expression and meaning of Nephrin and WT - 1 in podocyte of puromycin nephritis model

Objective To study the expression and meaning of Nephrin and WT - 1 in podocyte of puromycin nephritis model （PAN）. Methods Male adult SD rats were divided into control group and PAN group. PAN rats received a single intravenous injection of PAN via tail vein at 15mg/100g of body weight;control rats was injected by equal amount of 0.9% NS. The rats were executed at 4 weeks, 14 weeks and 20 weeks and the kidneys were taken out and embedded in paraffin for making the slices. The slices were stained using immunochemistry methods to detect the expression of nephrin and WT1, and anlysized by using semi - quantitative analysis. Results Theexpression of nephrin decreased slightly in PAN group in 4 weeks. （ 10.23 ± 1. 256 vs. 8.23 ± 0.88,P 〈 0.05 ） , and there is no expression of nephrin in segmental sclerosis in 14weeks （7.58±0.71 vs. 10.23 ±1.26,P 〈0.05） ,and in the sclerosis glomerular in 20weeks （7.88± 0.92 vs. 6.08 ± 1.34, P 〈 0.05 ） , compared with the control group showed significant differences. The expression of WT - 1 was no significant differences in 4 weeks, compared with the control （ 10.14± 2.02 vs. 10.11 ± 1.56,P 〉 0.05 ）. The expression of WT - 1 in PAN group decreased in 14 weeks （ 9.02 ± 1.73 vs. 6.8 ± 1.89, P 〈 0.05 ） , and decreased remarkably in 20 weeks （7.34 ± 1.22 vs. 4.43 ± 1.01, P 〈 0.05 ）. Conclusion The expressions of nephrin and WT - 1 are dropped according to the time course （from 4w to 20w）. These results indicate the degree of podocyte damage and the number losing of podocyte, therefore, the podo- cyte damage and number losing are relative to the glomerular sclerosis.