重组结缔组织生长因子对大鼠血管外膜成纤维细胞表型转化的影响

目的:克隆和构建带大鼠结缔组织生长因子(CTGF)基因真核表达载体pcDNA3.1(+)/CTGF,并观察其对血管外膜成纤维细胞(AF)表型的影响。 方法:以AF总RNA为模板，进行逆转录-聚合酶链反应(RT-PCR)，获得CTGF的cDNA，克隆入pcDNA3.1(+)载体，酶切和测序鉴定重组子。将构建好的pcDNA3.1(+)/CTGF用脂质体法转入AF细胞。Western blotting观察CTGF的表达。同时观察转染pcDNA3.1(+)/CTGF对细胞表型的影响。 结果:成功构建大鼠CTGF基因的真核表达载体pcDNA3.1(+)/CTGF．并证明其能在真核细胞内表达。转染pcDNA3.1(+)/CTGF可以诱导AF表型转化。 结论:重组结缔组织生长因子可诱导大鼠血管外膜成纤维细胞表型转化。

Effect of recombinant connective tissue growth factor on the phenotype transition of rat adventitia fibroblasts

AIM:To construct pcDNA3.1(+)/connective tissue growth factor (CTGF) eukaryotic expression plasmid and to investigate its role in the promotion of phenotypic transition in adventitia fibroblasts (AF). METHODS:The expression vector pcDNA3.1(+)/CTGF was constructed by routine molecular biological method. The expression vector pcDNA3.1(+)/CTGF was confirmed by restriction enzyme digestion and sequencing method. The expression vector pcDNA3.1(+)/CTGF was transfected into AF and the exogenous expression was observed. The expression of the α-SM 〖JP+1〗actin was examined by Western blotting. RESULTS:The eukaryotic expression vector of CTGF was successfully constructed, which was transfected into AF, the expressed CTGF promoted phenotype transition in AF. CONCLUSION:The pcDNA3.1(+)/CTGF plasmid was constructed and transfected into AF, the expressed CTGF promoted phenotype transition in AF.