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目的 观察 podocin在大鼠氨基核苷嘌呤霉素肾病模型中的表达和分布的改变 ,探讨其在蛋白尿发生中的可能作用。方法 通过一次性腹腔注射氨基核苷嘌呤霉素 (PAN)建立大鼠肾病模型 ,分别于注射后 1,3,10 ,2 0d处死大鼠 ,每次 6只。对照组注射等量的生理盐水。应用光镜、电镜观察肾脏病理改变 ,应用免疫荧光染色结合图像分析、半定量RT PCR的方法 ,检测肾组织的podocin的表达。结果 ①PAN注射后第 3天 ,大鼠 2 4h尿蛋白的排泄量逐渐增加 ,第 10天达高峰 ,较对照组差异有显著性 (P <0 .0 1) ;第 2 0天 ,模型组大鼠 2 4h尿蛋白排泄逐渐恢复 ,但仍高于对照组 (P <0 .0 5 )。②PAN肾病模型第 3、10天 ,透射电镜显示足细胞足突融合。③与对照组比较 ,肾小球podocin的表达在PAN注射后第 1天出现下调 ,第 3、10天显著下调 ,第 2 0天podocin的表达逐渐恢复 ,但仍低于对照组 (P <0 .0 1)。④Podocin在正常大鼠肾小球沿毛细血管襻 ,呈均匀连续的线样分布。肾病模型第 1天 podocin的分布变得不均匀 ,局部呈颗粒状分布 ;第 3天 podocin的分布呈现弥散性的颗粒状 ;第 10天podocin呈粗大的颗粒状分布。第 2 0天podocin的分布逐渐恢复为线样。⑤肾病模型第 1、3、10天PodocinmRNA水平较对照组表达略有增强 ,第 2 0天恢复
Objective To observe the expression and distribution of podocin in rat model of puromycin nephropathy and to explore the possible role of podocin in the pathogenesis of proteinuria. Methods A rat model of nephropathy was established by intraperitoneal injection of puromycin (PAN). Six rats were killed at 1, 3, 10 and 20 days after injection respectively. The control group was injected with the same amount of saline. The pathological changes of kidney were observed with light microscope and electron microscope. The expression of podocin in renal tissues was detected by immunofluorescence staining combined with image analysis and semi-quantitative RT-PCR. Results ① On the third day after PAN injection, urinary protein excretion increased 24 hrs and reached the peak on the 10th day, which was significantly different from the control group (P <0.01). On the 20th day, 24 h urinary protein excretion gradually recovered, but still higher than the control group (P <0. 05). ②PAN nephropathy model 3,10 days, transmission electron microscopy showed podocyte foot process fusion. ③Compared with the control group, the expression of podocin in glomerulus was down-regulated on the first day after PAN injection, significantly down-regulated on the 3rd and 10th days, podocin expression gradually recovered on the 20th, but still lower than that of the control group (P <0 .0 1). ④ Podocin in normal rat glomerular capillary loop, was uniform and continuous line-like distribution. On the first day, the distribution of podocin in the model of nephropathy became uneven, and the distribution of podocin was locally granular. The distribution of podocin was diffuse granular on the third day and the coarse granular distribution of podocin on the 10th day. On day 20, the distribution of podocin gradually recovered to a linear pattern. (5) The expression of Podocin mRNA on the 1st, 3rd and 10th day in nephropathy model increased slightly compared with the control group, and recovered on the 20th day