转染人TGF-β1基因对大鼠系膜细胞基质表达的影响  

Expression of ECM Components Protein of Cultured Rat Mesangial Cell by Transfecting Human TGF-β1 Gene

作　　者：刘国元[1] 蒋涛[1] 曾文姣[1] 刘学光[1] 赵仲华[1] 张农[1] 

机构地区：[1]复旦大学上海医学院病理学系,上海200032

出　　处：《复旦学报（医学版）》2004年第1期9-11,F002,共4页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(30170430)

摘　　要：目的 通过对大鼠系膜细胞(MsC)转染人TGF-β1基因,观察该基因对MsC表达细胞外基质成分的影响。方法 采用脂质体介导法转染人TGF-β1基因至大鼠MsC,Western blot法鉴定;细胞爬片ABC免疫酶标法观察纤维连接蛋白(FN)、Ⅳ型胶原、Ⅰ型胶原、层黏蛋白(LM)的表达,并对结果定量分析。结果 重组质粒pcDNA 3.0-TGF-β1成功转染至大鼠MsC,Western blot及免疫组化检测均证实TGF-β1蛋白表达增强;与正常的MsC相比,TGF-β1转染的MsC表达FN、Ⅳ型胶原增强,其差异有显著性意义;而Ⅰ型胶原、LM表达改变不明显。结论 TGF-β1可通过对大鼠MsC ECM成分表达的影响,而在肾小球硬化的发生发展过程起着重要作用。Purpose: To study the effect of transforming growth factor-β1 (TGF-β1) on protein expression of extracellular matrix components of mesangial cell by transfecting human TGF-β1 gene. Methods: Human TGF-β1 gene was transferred into mesangial cells (MsC) by lipofectin and confirmed by Western blot. Immunohistochemistry and image analysis were performed to detect the expression of type IV collagen, FN (fibronectin), type I collagen, LM (laminin). Results: Reconstructed plasmid pc-DNA3. 0-TGF-β1 was successfully transfected into MsC, Western blot and immunohistochemistry detection showed the protein of TGF-β1 was highly expressed in transfected cells. Immunohistochemistry and image analysis showed that the expression of type IV collagen and FN protein in transfected cells were much higher than that in the control group (P<0.01), but the difference of type I collagen and LM expression is not significant. Conclusions: TGF-β1 might play an important role in the processing of glomerulosclerosis through increasing synthesis of extracellular matrix(ECM) components protein in MsC.

关 键 词：基因转染 大鼠 系膜细胞 基因表达 转化生长因子Β1 纤维连接蛋白 Ⅳ型胶原 肾小球硬化 细胞外基质 

分 类 号：R692.6[医药卫生—泌尿科学] R394[医药卫生—外科学]