转染结缔组织生长因子基因的大鼠肾系膜细胞纤连蛋白和Ⅳ型胶原表达的改变

Changes of fibronectin and type Ⅳ collagen expression in cultured rat mesangial cells transfected with connective tissue growth factor expression vector

目的观察大鼠肾系膜细胞(MsC)转染结缔组织生长因子(CTGF)基因转染阳性细胞克隆中纤连蛋白(FN)和Ⅳ型胶原(Col Ⅳ)表达的改变,探讨 CTGF 在肾小球硬化发生中的作用。方法经脂质体介导将含有 CTGF 的重组表达质粒转染至大鼠 MsC,用 G418筛选及 Western blot 和半定量逆转录聚合酶链反应作鉴定;检测阳性克隆 FN、Col Ⅳ蛋白及其 mRNA 表达的改变。结果成功建立高表达 CTGF 的阳性 MsC 克隆(MCT-1,2),并证实其与对照组相比,阳性 MsC 克隆 FN、Col Ⅳ的表达均有上调,其 FN 蛋白及其 mRNA 表达分别增高2.9倍和3.2倍,Col Ⅳ蛋白和 mRNA 表达分别增高为2.4倍和3.8倍。结论 CTGF 可促进 MsC 的 FN、Col Ⅳ的表达,表明其在肾小球硬化发生中起促进作用。

Objective To study the role of connective tissue growth factor （CTGF） in the development of glomerulosclerosis by experimental alteration of fibronectin （FN） and Type Ⅳ collagen （ Col Ⅳ） expression in cultured rat mesangial cells （MsC）. Methods CTGF expression vector was transfected into MsC by Lipofectimine method. Protein and mRNA expression levels of CTGF, FN and Col Ⅳ were studied by Western blot and reverse transcription-polymerase chain reaction （RT-PCR） respectively. Results Two of MsC clones （ MCT-1 and MCT-2） with CTGF overexpression were successfully established and found to have significant increases of FN and Col Ⅳ at both protein and mRNA levels. Compared with the controls, the expression of FN protein and mRNA in the two clones were 3.2 times （ P 〈 0.05 ） and 2.9 times （ P 〈 0. 05 ） higher respectively. The expression of Col 1V protein and mRNA was 3.8 times （ P 〈 0. 01 ） and 2. 4 times （ P 〈 0. 01 ） higher respectively. Conclusion CTGF up-regulates FN and Col Ⅳ expression in MsC and may play an important role in the development of glomerulosclerosis.