Smad7基因转染拮抗TGF-β_1对培养肾小管细胞的影响

Transfection and expression of Smad7 inhibits transforming growth factor-β_1 effects on renal tubular cells

目的 探讨Smad7基因转染对TGF β1 诱导的小管细胞生长阻滞 ,细胞凋亡和FN分泌的影响。方法 采用Tfx 5 0脂质体将小鼠Smad7转染入原代培养的肾小管细胞 ,用MTT法观测小管细胞的增殖、流式细胞仪观测其细胞周期的变化 ;ELISA法测定小管细胞FN的分泌。结果 培养基添加TGF β1 ( 10ng ml) 48h后 ,小管细胞增殖能力明显下降 ,停滞于G1 期的细胞数增多 ,细胞分泌的FN量也明显增高 ,相反 ,Smad7基因转染后可明显拮抗TGF β1 对小管细胞的上述作用。结论 Smad7基因转染可明显拮抗TGF β1 对小管细胞的影响 。

Objective To explore the effect of Smad7 gene transfer on tubular cell cycle arrest, tubular apoptosis and fibronectin (FN) synthesis by transforming growth factor β 1(TGF β 1) induced. Methods Mouse Smad7 gene was transfected into renal tubular cells in primary cell culture by using Tfx 50 cationic liposome. Tubular cell proliferation was measured by MTT and cell cycle was observed by flow cytometry. The level of FN secretion in the supernatant was determined by ELISA. Results At 48 h after administration of 10 ng/ml TGF β 1 to renal tubular cells, cell proliferation declined, G 0/G 1 arrested in cell cycle, and cell FN secretion increased significantly. These abnormalities were attenuated by liposome mediated Smad7 gene transfection. Conclusion Transfection and expression of Smad7 can markedly inhibit TGF β 1 responses in renal tubular cells, which is helpful for further studies of Smad7 gene therapy in vivo .