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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 observe the changes of fibronectin (FN) and type Ⅳ collagen (Col Ⅳ) expression in rat mesangial cells (MsC) transfected with connective tissue growth factor (CTGF) The role of ball hardening occurs. Methods Recombinant plasmids containing CTGF were transfected into MsC by lipofectamine and identified by G418 selection, Western blot and semiquantitative reverse transcription polymerase chain reaction (RT - PCR). The positive clones FN and Col Ⅳ protein and mRNA were detected Changes in expression. Results Positive MsC clones (MCT-1, 2) highly expressing CTGF were successfully established and their expression of FN and Col IV were up-regulated in positive MsC clones compared with the control group. The expression of FN protein and its mRNA were increased by 2.9 Fold and 3.2 fold, respectively. Col Ⅳ protein and mRNA expression increased 2.4 fold and 3.8 fold, respectively. Conclusion CTGF can promote the expression of FN and Col Ⅳ in MsC, which indicates that CTGF can promote the occurrence of glomerulosclerosis.