DM大鼠肾组织Id2蛋白表达变化与纤维化病变发生发展关系的研究  

Study on the Relationship of Changes of Id2 Protein Expression in Diabetic Rat Kidney Tissues with Occurrence and Development of Fibrosis Lesions

作　　者：文箐颍[1] 苏博[1] 肖瑛[1] 王圆圆[1] 李霜[1] 刘丽荣[1] 石明隽[1] 郭兵[1] 

机构地区：[1]贵阳医学院病理生理学教研室,贵州贵阳550004

出　　处：《贵阳医学院学报》2015年第4期330-336,共7页Journal of Guiyang Medical College

基　　金：国家自然科学基金(81360116);贵州省科学技术基金[编号:黔科合J字(2011)2120号]

摘　　要：目的：动态观察分化抑制因子2（Id2）在糖尿病大鼠血糖控制前后肾小管上皮细胞的表达变化,探讨其在肾小管上皮细胞向间充质细胞转分化过程（EMT）中可能发挥的作用。方法：雄性SD大鼠随机分为正常对照组（N）、糖尿病组（DM）,糖尿病胰岛素治疗组（DMT）,采用链脲佐菌素（55 mg/kg）尾静脉注射复制Ⅰ型糖尿病大鼠模型,N组和DM组成模后分别于2周（2 w）、8周（8 w）、16周（16 w）、24周（24 w）时各处死6只;成模13周（13 w）起,DMT组大鼠采用胰岛素个体化治疗,以血糖控制在4-7 mmol/L,尿糖阴性为准,分别于16 w和24 w时处死6只大鼠;观察大鼠血糖、24 h尿蛋白并计算肾脏指数,HE、PAS染色光镜下观察肾组织结构的变化,免疫组化观察大鼠肾脏纤维连接蛋白（FN）的蛋白表达情况,Western blotting检测大鼠肾组织分化抑制因子（Id2）、E-钙粘素（E-cadherin）和α-平滑肌肌动蛋白（α-SMA）的蛋白变化,RT-PCR检测肾组织Id2 mRNA表达情况。结果：（1）与N组相比,不同时间点DM组血糖、24 h尿蛋白量、肾脏指数均显著升高（P〈0.01）,并出现肾组织形态学异常改变;而DMT组血糖、24 h尿蛋白量、肾脏指数均低于DM组（P〈0.05）,肾纤维化病变改善;（2）与N组比较,DM组肾组织Id2蛋白和mRNA的表达从2 w开始减少,16 w、24 w显著减少（P〈0.05）,并伴有E-cadherin的表达减少（P〈0.05）、α-SMA表达增加（P〈0.05）和FN蛋白的沉积增多（P〈0.05）,相关性分析显示Id2蛋白与FN蛋白成负相关（r=-0.931,P〈0.05）,与E-cadherin蛋白成正相关（r=0.900 0,P〈0.05）;（3）与DM组相比,DMT组大鼠肾组织Id2蛋白和mRNA表达均明显增加（P〈0.05）,且E-cadherin的表达增加（P〈0.05）、α-SMA的表达及FN在间质沉积减少（P〈0.05）。结论：胰岛素在控制血糖能上调肾小管上皮细胞Id2蛋白的表达,恢复Id2对肾纤维化的负调节作用,进而逆转�Objective：To investigate the expressions of Id2 in the renal tubules of STZ-induced dia-betic rats before and after blood glucose control,and explore their roles and relationship in the devel-opment of diabetic nephropathy.Methods：SD rats were randomly divided into control （N）,diabetes mellitus （DM）and insulin-treated DMgroups （DMT）.DMmodel was constructed by injecting strep-tozotocin （STZ）via tail vein （55 mg/kg）.48 SD rats were randomly divided into 4 groups：2 weeks, 8 weeks,16 weeks and 24 weeks group.Each group included a control subgroup and a diabetic model subgroup.In addition,for 16 weeks group and 24 weeks group,each group still included an insulin-treated subgroup.Since the 13 th week after modeling,rats in DMT group were given insulin individual- ly.The blood glucose was controlled to the level of 4 -7 mmol /L,and urine glucose remained nega-tive.The blood glucose,24 h urine protein were measured by biochemical method,and the kidney in-dex was measured as well.And the renal fibrosis was examined by HE and PAS staining sections.Im-munohistochemistry was employed to assay the expression of FN protein in the rat renal tissue.Western blotting was employed to assay the expression of Id2,E-cadherin and α-SMA protein in the rat renal tissue.In addition,the expression of Id2 mRNA was also examined by RT-PCR.Results：（1）Com-pared with group N,the blood glucose,24 h urine protein and the kidney index of group DMincreased significantly （P 〈0.01 ）,and renal tissue presented typical morphologic changes at different time points.By comparison,the blood glucose,24 h urine protein and the kidney index of group DMT were significantly lower than those in group DM（P 〈0.05）.Lesions of renal fibrosis in DMT rats were ob-viously relieved.（2）The expression levels of Id2 protein and mRNA of group DM decreased signifi-cantly than that in N group at different time points （P 〈0.05）,accompanied by the decreased expres-sion of E-cadherin （P 〈0.05）,increased expression of �

关 键 词：糖尿病肾病 纤维化 分化抑制因子2 纤维连接蛋白 E-钙黏素 Α-平滑肌肌动蛋白 大鼠 Sprague-Dawley 

分 类 号：R363[医药卫生—病理学]