基础医学与临床 ›› 2016, Vol. 36 ›› Issue (10): 1388-1392.

• 研究论文 • 上一篇    下一篇

蟾蜍灵抑制TGF-β1诱导人肾小管上皮细胞-间充质细胞系转换

郭静1,李善文2,施会敏2,张爱青2   

  1. 1. 南京医科大学
    2. 南京医科大学第二附属医院
  • 收稿日期:2015-07-23 修回日期:2016-02-28 出版日期:2016-10-05 发布日期:2016-09-27
  • 通讯作者: 张爱青 E-mail:njaiqing@njmu.edu.cn
  • 基金资助:
    南京医科大学科技发展基金重点项目;南京医科大学科技发展基金重点项目;南京市医学科技发展项目

Bufalin attenuates TGF-β1-induced epithelial-mesenchymal transition in HK2 cells

  • Received:2015-07-23 Revised:2016-02-28 Online:2016-10-05 Published:2016-09-27

摘要: 目的 探讨蟾蜍灵对转化生长因子(TGF)-β1诱导的人肾小管上皮细胞-间充质细胞转换的影响。方法 将体外培养的人近端肾小管上皮细胞系(HK-2)细胞分为3组:1)对照组;2)TGF-β1组:在细胞培养基中加入TGF-β1(浓度为5 μg/L);3)蟾蜍灵+TGF-β1组:在细胞培养基中分别加入蟾蜍灵和TGF-β1(浓度为5 μg/L),按蟾蜍灵的浓度分为A、B、C 3个亚组:分别为1×10-9、1×10-8和1×10-7mol/L。72 h后,用倒置相差显微镜观察细胞形态;用实时定量PCR、蛋白印迹法和免疫荧光检测细胞α-平滑肌肌动蛋白(α-SMA)和E-钙黏蛋白(E-cadherin)的表达。结果 TGF-β1组HK-2细胞从原有典型的铺路石样上皮细胞转变为长梭形肌成纤维细胞;胞质内大量表达α-SMA,同时E-cadherin的表达明显减少(P<0.01);蟾蜍灵处理后TGF-β1诱导的细胞形态学改变减轻,胞质内α-SMA的表达减少(P<0.05),同时E-cadherin的表达明显恢复,且呈剂量依赖性。结论 蟾蜍灵能有效抑制TGF-β1诱导的HK-2细胞转换,对肾脏病治疗具有潜在应用前景。

关键词: 蟾蜍灵, 转化生长因子-β1, 上皮细胞-间充质细胞转换

Abstract: Objective To investigate the preventive effects of bufalin on transforming growth factor-beta 1(TGF-β1) induced epithelial-mesenchymal transition of human kidney cells. Methods Human kidney proximal tubular cell line (HK-2)was used as the proximal tubular cell model. Cells were divided into five groups as follows:control group, TGF-β1 (5 μg /L) group, TGF-β1 (5 μg/L) plus bufalin (1×10-9、1×10-8 and 1×10-7mol/L) groups. Epithelial to mesenchymal transition(EMT) was induced with 5 μg/L of human TGF-β1. The effects of bufalin on cell morphology were observed by phase contrast microscopy, the expression of α-smooth muscle actin (α-SMA) and E-cadherin were measured by immunofluorescent staining, real-time reverse transcription-polymerase chain reaction and western blot. Results Our results revealed that bualin not only prevented α-SMA expression(P<0.05) but also prohibited the decrease of the epithelial marker E-cadherin(P<0.05) in HK-2 cells in a dose-dependent manner. Simultaneous incubation of bufalin with TGF-β1 could protect the change to the myofibroblast phenotype and restore the epithelial morphology of the HK-2 cells. Conclusion These observations strongly suggest that bufalin is a potent inhibitor of TGF-β1-induced EMT and may be a promising agent for treating tubulointerstitial fibrosis.

Key words: bufalin, transforming growth factor-β1, epithelial-mesenchymal transition

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