[1]赵礼金,杨日高,程龙,等.肝内胆管上皮细胞上皮-间叶转化的实验研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(03):214-219.
 Zhao Lijin,Yang Rigao,Cheng Long,et al.Epithelial-mesenchymal transition of intrahepatic biliary epithelial cells[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(03):214-219.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第03期
页码:
214-219
栏目:
论著
出版日期:
2010-02-15

文章信息/Info

Title:
Epithelial-mesenchymal transition of intrahepatic biliary epithelial cells
作者:
赵礼金杨日高程龙王麦建江艳余德刚周廷梅王曙光
第三军医大学西南医院全军肝胆外科研究所;遵义医学院附属医院肝胆外科;解放军324医院普外科
Author(s):
Zhao Lijin Yang Rigao Cheng Long Wang Maijian Jiang Yan Yu Degang Zhou Tingmei Wang Shuguang
Department of Institute, Southwest Hospital, Third Military Medical University, Chongqing, 400038;Affiliated Hospital of Zunyi Medical College, Zunyi, Guizhou Province, 563003; Department of General Surgery, No. 324 Hospital of PLA,  Chongqing, 400020, China
关键词:
肝内胆管上皮细胞表型脂多糖类Smad蛋白类转化生长因子-β1紫杉醇RNA 小分子干扰
Keywords:
bile ducts intrahepaticepithelial cellsphenotypelipopolysaccharidesSmad proteinstransforming growth factor beta 1paclitaxel RNA small interfering
分类号:
R-33;R575.702;R394.3
文献标志码:
A
摘要:
目的  观察人肝内胆管上皮细胞(human intrahepatic biliary epithelial cells,HIBEpiC)上皮-间叶转化(epithelial-mesenchymal transitions, EMT)现象,探讨EMT在胆管周围纤维化中的作用及其可能的分子机制。  方法  HIBEpiC用脂多糖(lipopolysaccharide,LPS)处理48、72 h后,用PCR及Western blot技术检测E-cadherin、S100A4和α-SMA的表达,以及与EMT信号通路相关的TGF-β1的表达;同时用紫杉醇、siRNA smad2/3阻断TGF-β1的作用,探讨TGF-β1/smad2/3信号通路在HIBEpiC发生EMT时的可能作用。  结果  HIBEpiC经LPS处理后,TGF-β1 mRNA表达于48 h达到高峰,72 h后开始下降,但维持较高水平(P<0.01,P<0.05);EMT标志物E-cadherin mRNA和蛋白表达随培养时间的延长逐渐降低(P<0.01),而S100A4、α-SMA mRNA及蛋白表达明显上调(P<0.01,P<0.05)。紫杉醇可使HIBEpiC中LPS所诱导的TGF-β1、Smad2/Smad3 mRNA表达明显下降,特别是在48 h最明显(P<0.01)。用siRNA技术可使Smad2/3 mRNA及蛋白表达明显下调(P<0.01),同时可使与HIBEpiC共培养EMT标志物E-cadherin mRNA及蛋白质表达上调(P<0.01,P<0.05),S100A4和α-SMA表达明显下调(P<0.01)。  结论  LPS可诱导HIBEpiC中TGF-β1的表达,使胆管上皮细胞发生EMT,抑制TGF-β1或Smad2/3,可使EMT发生逆转,提示TGF-β1或Smad2/3可能成为预防胆道周围纤维化的一个潜在靶点。
Abstract:
Objective  To observe the epithelial-mesenchymal transition (EMT)of human intrahepatic biliary epithelial cells (HIBEpiC) and to study its role in bile duct fibrosis and its molecular mechanism.    Methods  Expressions of E-cadherin, S100A4, α-SMA, and EMT signal-related TGF-β1 in HIBEpiC were detected by RT-PCR and Western blotting 48 and 72 h after treatment of HIBEpiC with lipopolysaccharide (LPS). The effect of TGF-β1 was blocked with paclitaxel and siRNA smad2/3. The role of TGF-β1/ smad2/3 signal pathway in the occurrence of EMT in HIBEpiC was studied.    Results  The expression of TGF-β1 mRNA reached its  peak and began to decrease 48 and 72 h, respectively, after treatment of HIBEpiC with LPS , and still maintained a rather high level (P<0.01, 0.05). The expression level of EMT marker, E-cadherin mRNA and TGF-β1 decreased with the prolongation of culture time ( P<0.01) while the expression of S100A4, α-SMA mRNA and TGF-β1 significantly increased (P<0.01,0.05), suggesting that paclitaxel can down-regulate the LPS-induced expression of TGF-β1, Smad2 and Smad3 mRNA. siRNA could also down-regulate the LPS-induced expression of Smad2/3 mRNA, S100A4, α-SMA and TGF-β1 while up-regulate the expression of E-cadherin and TGF-β1 in HIBEpiC (P<0.01), indicating that EMT can be reversed.    Conclusion  LPS can induce expression of TGF-β1 in HIBEpiC, resulting in EMT in biliary epithelial cells, and reverse EMT by inhibiting TGF-β1 or Smad2/3, which may be a potential target for the prevention and treatment of  biliary fibrosis.

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更新日期/Last Update: 2010-01-27