炮制前后延胡索对TGF-β1介导上皮细胞-间充质转化作用的影响

Effect of corydalis rhizoma on TGF-β1-mediated EMT by processing

目的:研究炮制前后延胡索对转化生长因子(TGF-β1)介导的人膀胱上皮细胞ECV304上皮细胞-间充质转化(EMT)作用的影响。方法:以ECV304为实验对象,使用TGF-β1诱导细胞发生EMT,通过MTT法、细胞划痕、Western-blot、RT-PCR等实验考察延胡索对EMT的影响。结果:实验使用TGF-β1成功诱导ECV304细胞发生EMT;生延胡索6μg/L干预可恢复ECV304的细胞形态,并可显著抑制ECV304细胞的迁移能力,上调E-cadherin和下调Fibronectin蛋白表达,还能显著上调CDH1和下调FN1的mRNA表达,炮制后作用减弱。结论:延胡索对TGF-β1介导的EMT有抑制作用,生品抑制作用最强,作用机制与改变E-cadherin蛋白和Fibronectin蛋白的表达,及CDH1和FN1的mRNA表达有关。

Objective： To study the effect of corydalis rhizoma on TGF-β1-mediated EMT in ECV304 cells. Methods： The human bladder epithelial cells（ECV304） were used to investigate the effect of corydalis rhizoma on epithelial mesenchymal transition（EMT）,using transforming growth factor（TGF-β1） to induce the EMT of cells. The effect of corydalis rhizoma on EMT induced by TGF-β1 was studied by MTT,cell scratches,Western-blot and RT-PCR. Results： The EMT was successfully induced by TGF-β1 in ECV304 cells. The cell morphology of ECV304 cells could recover and the migration of ECV304 cells could be inhibited significantly interfered by raw corydalis rhizoma. Raw corydalis rhizoma could up regulation of E-cadherin and down regulation of Fibronectin protein expression,and also significantly up-regulated the mRNA expression of CDH1 and down-regulation of FN1. The effect was weakened after the processing. Conclusion： The corydalis rhizoma can inhibit TGF-β1-induced epithelial mesenchymal transition,and the inhibitory effect of raw material was the strongest. The mechanism of action is related to the changes in the expression of E-cadherin and Fibronectin protein,and the expression of mRNA in CDH1 and FN1.