Genistein对增生性瘢痕成纤维细胞TGF-β1表达及细胞内游离钙的影响

目的 观察5,7,4'-三羟基异黄酮(Genistein)对增生性瘢痕成纤维细胞TCF-β1的表达及细胞内钙离子浓度的影响,探讨Genistein抗纤维化作用的机制.方法 体外培养人增生性瘢痕成纤维细胞,以不同浓度Genistein(25、50、100μmol/L)处理,RT-PCR法检测药物作用48h后细胞TGF-β1mRNA的表达,Western Blot法检测细胞TGF-β1蛋白表达量的变化;激光共聚焦显微镜动态观测Genistein处理后成纤维细胞内Ca2+浓度以及Genistein预处理后再加bFGF刺激的细胞Ca2+浓度的动态变化.结果 Genistein作用后增生性瘢痕成纤维细胞表达TGF-β1的mRNA与蛋白量均显著下调,并具有剂量依赖性;bFGF可使培养的成纤维细胞内游离Ca2+浓度升高,经Genistein预处理的细胞内Ca2+的浓度明显下降.结论 Genistein能抑制增生性瘢痕成纤维细胞TGF-β1,的表达,拮抗生长因子促细胞内游离Ca2+浓度增加的效应,可能是其抗纤维化作用的机制之一.

Effects of genistein on expression of TGF-β1and Intracellular free CO2+Concentration in fibroblasts derived from human hypertrophic scar

Objective To observethe effects of Genistein on TGF-β1 expression and the intracellular free Ca2+ concentration in human hyperu'ophic scar fibroblasts,and to discuss the mechanism of the anti-fibrosis effect.Methods Fibroblasts were derired from human hypertrophic scar tissue and cultured in vitro.Genistein in different concentrations(25、50、100 μmol/L)was administrated to the fibmblasts,respectively.After 48 hours of co-culture,the expression of TGF-β1 mBNA and protein were examined by RT-PCR and Western-Blot assay respectively.The intracetlular free Ca2+ concentration in hypertrophic gear fibmblasts pretreated by Genistein was determined by laser confocal scanning microscopy with or without the stimulation of bFGF.Results Genistein inhibited the expression of TGF-β1 in hypertrophic scar fibroblasts on a concentration-dependent manner.bFGF significantly elevated the intracellular free Ca2+ concentration.however its stimulating effect was remarkably alleviated when the flbroblasts were pro-treated by Genistein.Conclusions Genistein can reduce the expression of TCF-β1 and block the accumulation of intmcellular free calcium induced by growth factors.It maybe one of the possible mechanisms of Genistein's antifibrosis effect.