回输转染饰胶蛋白聚糖基因的肾系膜细胞对大鼠抗thy-1血清性肾炎的拮抗作用  

The antagonistic effect on anti-thy-1 serum-induced nephritis of rats injected by decorin-transfected mesangial cells vector

作　　者：王慧君[1] 张志刚[1] 刘学光[1] 张秀荣[1] 陈琦[1] 郭慕依[1] 

机构地区：[1]复旦大学上海医学院病理学系,200032

出　　处：《中华病理学杂志》2003年第5期444-448,共5页Chinese Journal of Pathology

基　　金：国家自然科学基金资助项目 ( 31 0 70 4 31 );教育部博士点基金资助项目 ( 2 0 31 );上海市教委重点学科基金资助项目 (B990 80 2 )

摘　　要：目的　以转染饰胶蛋白聚糖 (decorin)基因的肾系膜细胞 (MsC)为载体 ,经肾动脉回输入抗thy 1血清性肾炎大鼠肾小球 ,观察其存活情况及对模型大鼠肾小球病变的影响及意义。方法　用脂质体介导法将载有目的基因 (decorin)片段转入正常MsC株而获得阳性表达之细胞克隆 ;经尾静脉注射兔抗大鼠thy 1血清 (ATS)复制其抗thy 1系膜增生性肾炎模型 ;经左肾动脉将转基因MsC回输入thy 1肾炎大鼠肾小球 ,并经该鼠MsC原代培养、BrdU和decorin蛋白免疫组织化学染色结合图像分析观察其生存情况 ;应用转化生长因子 β1(TGF β1)、纤连蛋白和Ⅳ型胶原免疫组织化学结合图像分析研究对thy 1系膜增生性肾炎动物模型肾小球病变的影响。 结果　经制备兔抗大鼠ATS ,将其从尾静脉注入大鼠体内成功复制thy 1系膜增生性肾炎模型。经左肾动脉直接注入转染decorin基因的MsC克隆 (D A6) ,经原代培养证实其生长活跃 ,用免疫组织化学法发现其细胞核和细胞质均可分别表达BrdU和decorin蛋白。与未经注射对侧肾相比 ,回输侧肾的肾小球TGF β1、纤连蛋白和Ⅳ型胶原的表达均降低 ,分别为TGF β14d时P <0 .0 5和纤连蛋白、Ⅳ型胶原 1～ 2d时P <0 0 1。结论　通过肾动脉直接输入法将转染decorin基因的细胞克隆回输入大鼠抗thy 1血清性肾炎肾小?Objectives To inject decorin-transfected mesangial cells (MsC) vector into the kidneys of rats with anti-thy-1 serum-induced nephritis via left renal artery and observe the survival condition of MsC vector and its influence on glomerular lesions in rats with anti-thy-1 serum induced nephritis. Methods Rat mesangio-proliferative glomerulonephritis was established by tail intravenous injection with rabbit anti-thy-1 serum (ATS). Decorin-transfected MsC was injected into ra t kidneys via left renal artery. Primary culture, immunostaining for BrdU and decorin of transfected MsC lines were performed to observe their survival. Immunohistochemistry with image analysis was performed to detect the expression of BrdU, α-SMA, decorin, TGF-β1、FN and ColⅣ in diseased glomeruli. Results Rat anti-thy-1 serum-induced nephritis identified by pathological examination was successfully established by injecting rabbit ATS, and decorin transfected MsC vector was transfused to rat glomeruli via left renal artery. The active growth and positive expressions of BrdU and decorin proteins on the nuclei and cytoplasms of ex vivo MsC were observed respectively. TGF-β1, FN, ColⅣ expressions in diseased glomeruli of rats with ATS nephritis were decreased significantly at day 4(TGF- β1 , P<0.05) and day 2 (FN and ColⅣ, P<0.01) respectively, compared to uninjected kidneys. Conclusions MsC vector is successfully transferred to the glomeruli of experimental rats via left renal artery injection with no affect on cell survival. Decorin protein is expressed on the transfected MsC and shows antagonistic effect on the glomerular lesions of ATS rats. It suggests that the use of ex vivo MsC vector system can provide useful experimental basis for gene therapy of kidney disease in animal model.

关 键 词：大鼠 回输 肾小球 肾炎 血清 转染 肾动脉 细胞克隆 目的基因 原代培养 

分 类 号：R692.3[医药卫生—泌尿科学]