TNF-冄激活大鼠胆管上皮细胞Jagged-1的表达并诱导其上皮-间质转化  

Tumor necrosis factoralpha up-regulates expression of Jagged-1 and induces epithelialmesenchymal transition in rat cholangiocytes in vitro

作　　者：李潼[1] 郭兵[1] 高杨[1] 于奇宏 李锦锦[1] 鲜文静 蒋帅[1] 郑启昌[1] 张勇[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院肝胆外科,湖北省武汉市430030 [2]华中科技大学同济医学院附属协和医院麻醉科,湖北省武汉市430030

出　　处：《世界华人消化杂志》2016年第12期1806-1811,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金面上项目;No.81570566;2015年度湖北省留学人员科技活动项目择优资助经费~~

摘　　要：目的:检测肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)对体外培养的原代大鼠肝内胆管上皮细胞Jagged-1表达的影响以及诱导其间质-上皮转化的作用.方法:原代胆管上皮细胞分为对照组,TNF-α处理组(10 ng/mL),TNF-α加核转录因子-κB(nuclear factor kappa B,NF-κB)抑制剂PDTC(pyrrolidine dithiocarbamate)(50冚mol/L)处理组及PDTC单独处理组,72 h后Western blot法检测各组细胞Jagged-1蛋白、间质细胞标记成纤维细胞特异性蛋白-1(fibroblast specific protein 1,FSP-1)、波形蛋白(Vimentin)、α-平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)和胆管细胞特异性标记角蛋白19片段(cytokeratin 19,CK19)的蛋白的表达,凝胶迁移或电泳迁移率实验(EMSA)检测NF-κB蛋白结合活性.倒置显微镜观察各组胆管细胞的形态变化,Transwell小室法检测细胞的迁移能力.结果:TNF-α处理组胆管上皮细胞NF-κB的活性增强,Jagged-1、FSP-1、Vimentin和α-SMA蛋白的表达水平上调,CK19的表达下调,细胞迁移能力增强,细胞形态由鹅卵石样向梭形样转化.TNF-α+PDTC与TNF-α组相比其NF-κB活性明显减弱,Jagged-1,FSP-1,Vimentin和α-SMA蛋白水平降低,CK19表达升高,细胞迁移能力减弱,细胞形态向鹅卵石样转化.结论:TNF-α可通过激活NF-κB信号上调Jagged-1蛋白表达的同时诱导大鼠胆管上皮细胞向间质细胞转化.AIM： To investigate the role of tumor necrosis factor-alpha （TNF-α in the epithelial-mesenchymal transition （EMT） process in rat cholangiocytes in vitro. METHODS： Primary rat cholangiocytes were treated with TNF-α （10 ng/mL） alone, TNF-α plus nuclear factor kappa B （NF-κB） inhibitor PDTC （50 μmol/L）, or PDTC alone for 72 h. The expression of Jagged-I, mesenchymal markers [fibroblast-specific protein-1 （FSP-1）, Vimentin and α-SMA] as well as epithelial marker CK19 was detected by Western blot. NF-κB binding activity was measured by EMSA. Migration ability and morphological changes of cholangiocytes were also examined. RESULTS： In the TNF-α alone group, the protein levels of Jagged-I, FSP-1, Vimentin and α-SMA were up-regulated compared to control cells, whereas the expression of CK19 was down-regulated. The migration ability of cholangiocytes was increased and their shape changed from stone-like to fiber-like. For the TNF-α plus PTCD group and the PTCD alone group, no significant changes in EMT markers as well as migration ability were observed compared to control cells. CONCLUSION： TNF-α is able to increase the expression of Jagged-1 and induce EMT in rat cholangiocytes in vitro possibly through activation of NF-κB signaling.

关 键 词：肿瘤坏死因子-冄 胆管上皮细胞 上皮-间质转化 

分 类 号：R57[医药卫生—消化系统]