TGFβ1反义RNA对腹膜间皮细胞分泌细胞外基质的影响

目的：研究转化生长因子β1（TGFβ1）反义RNA对体外培养的人腹膜间皮细胞（HPMC）细胞外基质合成与分泌的影响。 方法：将反义TGFβ1基因真核表达载体转染HPMC，用ELISA法检测转染细胞上清液中纤维连接蛋白（FN）和Ⅰ型纤溶酶原激活物抑制剂（PAI-1）的蛋白质水平；用半定量逆转录多聚酶链反应（RT-PCR）法检测转染细胞内FN，PAI-1和胶原Ⅰ（ColⅠ）的mRNA表达，并与未转染HPMC对照组和转染空载体组比较其变化。 结果：TGFβ1反义RNA转染HPMC后，与对照组相比，转染24 h后，FN，ColⅠ，PAI-1 mRNA分别下调17％，26％，9.6％；转染48ｈ后 FN，PAI-1蛋白含量亦明显下降（P<0.05）。 结论：人反义TGFβ1基因真核表达载体可抑制HPMC合成细胞外基质，在腹膜纤维化的研究及防治中有潜在的应用价值。

Effect of TGFβ1 antisense RNA expression vector on synthesis and secretion of extracellular matrix in cultured human peritoneal mesothelial cells

OBJECTIVE: To determine the effect of transforming growth factor beta1 (TGFbeta1) antisense RNA on the synthesis and secretion of extracellular matrix in human peritoneal mesothelial cells (HPMC). METHODS: The recombinant human TGFbeta1 antisense eukaryotic expression vector was transfected into HPMC. Fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) were detected by ELISA method. The expression of FN, collagen I (col I) and PAI-1 mRNA were detected by RT-PCR method. RESULTS: The mRNA of FN, Col I,nd PAI-1 in HPMC were repressed by TGFbeta1 antisense RNA 24 hours after the transfection in comparison with the control group, which decreased 17%, 26%, and 9.6%, respectively. The protein of FN and PAI-1 were inhibited 48 hours after the transfection (P < 0.05). CONCLUSION: Recombinant human TGFbeta1 antisense eukaryotic expression vector can inhibit the synthesis and secretion of extracellular matrix in HPMC which may be effective in gene therapy for peritoneal fibrosis.