Podocin与nephrin在多柔比星诱导肾病大鼠模型肾皮质部的表达及共定位分布  

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

作　　者：温捷[1] 于力[1] 温跃强[1] 郝志宏[1] 王丽娜[1] 陈蓉燕[1] 

机构地区：[1]广州医学院附属广州市第一人民医院儿科,广州510180

出　　处：《中华妇幼临床医学杂志（电子版）》2009年第4期1-5,共5页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：广东省自然科学基金项目(No.5000424);广州市科技攻关计划项目资助(2006Z3-E0231);番禺区科技计划项目(2007-Z-78-1)~~

摘　　要：目的观察podocin与nephrin在多柔比星诱导肾病(adriamycin-inducing nephrotic,ADN)大鼠模型病变过程中的表达及共定位分布,探讨podocin和nephrin在肾病病变过程中的作用机制。方法将60只雄性Spraque-Dawley(SD)大鼠分为模型组(尾静脉注射6.5mg/kg多柔比星,n=30)和对照组(尾静脉注射0.8mL生理盐水,n=30)。分别于造模后的第1、第2、第4、第6和第8周末处死大鼠,并观察各项指标的变化:①24h尿蛋白定量,血浆白蛋白、血胆固醇、血肌酐和血尿素氮水平;②两组上述各时间点取肾皮质进行光镜和电镜检测;③采用双标准曲线法进行荧光定量PCR检测肾皮质部podocin mRNA和nephrin mRNA表达;④应用双重免疫荧光法观察肾小球podocin和nephrin的共定位分布。结果①模型组大鼠出现大量蛋白尿、低白蛋白血症、高脂血症,肾功能减退,表现为典型的肾病综合征;模型组病理改变由微小病变逐渐发展至局灶节段性肾小球硬化,电镜下可见足细胞在病变早期出现足突增宽、融合,晚期足突消失、细胞核固缩;②模型组podocin mRNA表达第1周末时升高95.8%(P<0.01),第2周末时升高252.5%(P<0.01)且达高峰,第4周末时开始下降27.6%(P<0.05),第6和第8周末分别下降52.9%和48.0%,与对照组比较,差异有显著意义(P<0.01)。模型组nephrin mRNA表达第1周末时升高140.8%(P<0.05)且达高峰,第2周末时持续升高80.4%(P<0.05),第4、第6和第8周末时分别下降39.2%,47.5%和52.5%,与对照组比较,差异有显著意义(P<0.01)。③模型组免疫荧光染色可见podocin与nephrin在大鼠肾小球内分布,且随病变进展而增加,由正常的沿肾小球血管襻呈均匀线性分布转变为呈不连续分布,而且重叠程度减少。结论Podocin和nephrin在多柔比星肾病大鼠肾病进展过程中出现表达及分布异常,导致两者不能形成正常的复合体结构,这可能是蛋白尿发生的重要分子机制。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 iime 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 （1） ADN 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

关 键 词：PODOCIN NEPHRIN 多柔比星诱导肾病 肾病综合征 蛋白尿 

分 类 号：R692.9[医药卫生—泌尿科学]