摘要
目的探讨表没食子儿茶素没食子酸酯(EGCG)对肾小管上皮细胞转分化的作用机制。方法培养大鼠肾小管上皮细胞株NRK-52E,将其分成4组。1组:正常对照组(N),以DMEM培养液为空白对照。2组:转化生长因子(TGF-β1)诱导组;3组:EGCG200μg/L干预组;4组:EGCG400μg/L干预组。各细胞组在作用48 h后,应用免疫组化检测平滑肌肌动蛋白(α-SMA)和角蛋白(CK-18),Western-blot分析胞浆蛋白细胞周期素D1(Cyclin D1)、基质金属蛋白酶9(MMP9)含量,实时荧光定量PCR法测Cyclin D1、MMP9mRNA的相对表达量。结果 NRK细胞被TGF-β1诱导48 h后,胞浆中出现α-SMA蛋白高表达,CK-18蛋白表达明显减少;Cyclin D1、MMP9蛋白及其mRNA出现了高表达;EGCG干预组上述改变明显减轻。结论 EGCG以剂量依赖方式抑制TGF-β1诱导的NRK细胞的转分化,其机制可能是通过抑制Cyclin D1、MMP9蛋白及mRNA表达而实现的。
Objective To explore the effect and mechanism of epigallocatechin-3-gallate（EGCG）on the transdifferentiation of renal tubular epithelial cells.Methods Tubular epithelial cell of rat（NRK-52E）was cultured and then were divided into four groups.Group one：Normal control（N）,Group two：TGF-β1 induction group,Group three：200 μg/L EGCG experimental group,Group four：400 μg/L EGCG experimental group.The protein expression of α-SMA,CK-18 were detected by immunostaining,cyclin D1,MMP9 were detected by Western-blotting,and the mRNA expression of cyclin D1,MMP9 from cell line were detected by real-time PCR at 48 hours after treatment.Results When the NRK cells were induced by TGF-β1 for 48 hours,the cytoplasmic protein α-SMA began expression and the expression of CK-18 reduced,the protein and mRNA expression of cyclin D1,MMP9 were up-regulated,such changes in the EGCG group were slight.Conclusion EGCG can inhibit the transdifferentiation induced by TGF-β1 of NRK cells in a concentration-dependent manner.The mechanism may be the inhibition of protein and mRNA expression of MMP9 and cyclin D1.
出处
《实用药物与临床》
CAS
2011年第3期185-188,共4页
Practical Pharmacy and Clinical Remedies
基金
沈阳市科学技术计划项目(F10-205-1-29)
中国医科大学附属盛京医院科研基金
