Chinese Medical E-ournals Database

Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition) ›› 2009, Vol. 05 ›› Issue (04): 331 -337. doi: 10.3877/cma.j.issn.1673-5250.2009.04.101

Original Article

Expression and Co-Localization of Nephrin and Podocin in Adriamycin-Induced Nephrotic Rats

Jie WEN, Li YU, Yue-qiang WEN, Zhi-hong HAO, Li-na WANG, Rong-yan CHEN   

  1. Department of Pediatrics, Affiliated Guangzhou First Municipal People's Hospital of Guangzhou Medical College, Guangzhou 510180, China
  • Published:2009-08-01
  • Supported by:
    * Project No.5000424, supported by the Natural Science Foundation Project of Guangdong Province; project No.2006Z3–E0231, supported by the Science and Technology Research Project of Guangzhou; project No.2007–Z–78–1, supported by the Science and Technology Research Project of Panyu District
Objective

To observe the expression and distribution of nephrin and podocin in adriamycin-inducing nephrotic (ADN) rats, and explore the possible mechanisms of nephrin and podocin during the progression of nephrotic syndrome(NS).

Methods

Sixty Spraque-Dawley(SD) rats were divided into ADN model group(injected 6.5 mg/kg adriamycin in tail vein, n=30)and control group (injected 0.8 mL saline solution in tail vein, n=30). After the nephropathy model was established, rats were killed at the end of 1st, 2nd, 4th, 6th, 8th week. The changes of the following indicators were observed: ① 24-hour urinary protein, serum albumin, serum cholesterol, serum creatinine and serum urea nitrogen were detected; ② The characters of renal cortex detected by optical microscope and electron microscope at different time points were observed; ③ The expression of podocin mRNA and nephrin mRNA in cortex of kidney were examined by real time RT-PCR; ④ The co-localization of nephrin and podocin in glomerular was observed by dual-labeling immunofluorescence.

Results

①ADN model group came out massive proteinuria, hypoalbuminemia, hypercholesterolaemia, renal failure, and presented as nephrotic syndrome. ADN model group developed from minimal change diseases to focal segmental glomerulosclerosis. The ultrastructure of podocyte showed that foot process conflurence in the early stage, foot process disappeared and chromatin raritas in the end stage. ② ADN model group compared with control group: podocin mRNA expression of AND model group increased 95.8% (P<0.01)at the end of the first week, increased 252.5%( P<0.01) at the end of the second week and reached the peak value, then began to decrease 27.6%(P<0.05)at the end of the fourth week, and decreased 52.9%(P<0.01)at the end of the sixth week, and decreased 48.0%(P<0.01)at the end of the eighth week. Nephrin mRNA expression of ADN model group increased 140.8% (P<0.05) at the end of the first week and reached the peak value, then increased 80.4%( P<0.05)at the end of the second week, decreased 39.2%(P<0.01)at the end of the fourth week, decreased 47.5%(P<0.01)at the end of the sixth week, decreased 52.5%(P<0.01)at the end of the eighth week. ③In ADN model group, the distribution of nephrin and podocin gradually shifted from a linear-like pattern along the capillary loops of glomerulus to a discontinuous coarse granular pattern, and the degree of overlapping localization decreased.

Conclusion

The abnormal expression and distribution of nephrin and podocin during the progression of adriamycin nephropathy lead nephrin and podocin can not form normal complex, but it may be the important molecular mechanism of proteinuria.

图1 两组肾组织病理学变化[图A、图B、图C、图D分别为对照组及模型组第4、第6、第8周末的肾小球组织结构(光镜×400);图E、图F、图G、图H分别为对照组及模型组第4、第6、第8周末的足细胞及足突超微结构(电镜×15 000)]
图2 Podocin、nephrin和β-actin琼脂糖凝胶电泳图
图3 两组nephrin mRNA表达变化
图4 Podocin和nephrin在肾小球的共定位分布(免疫荧光×400)[图A、图D、图G、图J、图M分别为对照组,模型组第2、第4、第6、第8周末podocin和nephrin在肾小球共定位分布;图B、图E、图H、图K、图N分别为对照组,模型组第2、第4、第6、第8周末nephrin在肾小球的分布;图C、图F、图I、图L、图O分别为对照组,模型组第2、第4、第6、第8周末podocin在肾小球的分布]
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