IL-6在人胆管上皮细胞损伤修复中的作用

蒋桂星; 崔云甫; 曹利平; 邰升; 钟翔宇; 王志东

doi:10.3760/cma.j.issn.1673 9752.2012.05.019

摘要: 目的探讨 IL6在人胆管上皮细胞(BECs)损伤修复中的作用机制。方法 IL6按浓度分为5组:0ng/L组,10ng/L组,50ng/L组,100ng/L组,1000ng/L组,将不同浓度的 IL6分别干预体外培养的 BECs,并检测 IL6对转录激活子3(STAT3)磷酸化和三叶因子3(TFF3)表达的影响;将 BECs分为未处理组、STAT3RNAi组(细胞中转入 STAT3RNAi腺病毒)和 ControlRNAi组(细胞中转入空 RNAi腺病毒),检测行干扰后,IL6对 TFF3表达的影响;分别用未处理组、STAT3RNAi组、ControlRNAi组 BECs制备体外损伤模型,观察 IL6、TFF3对各组细胞的影响。两组数据间比较采用 Student′st检验,多组间比较采用单因素方差或 Sidak检验。结果添加 IL6的 50ng/L组、100ng/L组、1000ng/L组磷酸化 STAT3 (pSTAT3)的表达水平分别为0.240±0.052、0.714±0.124、0.327±0.069,明显强于0ng/L组的0.033± 0.011(q=5.246,17.260,7.451,P<0.05)。TFF3mRNA及其蛋白表达水平随着 IL6浓度的增高而显著增加(q=12.045,9.889,P<0.05);IL6浓度为 1000ng/L时,TFF3mRNA及其蛋白表达有所回落。行干扰后,STAT3RNAi组 BECsTFF3蛋白表达水平为 0.037±0.005,明显低于 ControlRNAi组的 0.267± 0.038和未处理组的0.301±0.042(q=12.135,13.929,P<0.05)。体外损伤模型中,IL6干预 BECs12h 后,STAT3RNAi组 BECs移行速度为(9.1±1.5)μm/h,慢于 ControlRNAi组的(25.1±3.8)μm/h(q= 7.737,P<0.05);而 STAT3RNAi组中加入外源性人重组 TFF31g/L后,BECs移行速度为(39.2±4.7)μm/h, 比 ControlRNAi组显著提高(q=14.507,P<0.05)。结论 IL6主要通过激活 STAT3,继发上调 TFF3表达,从而促进人胆管上皮细胞移行和损伤修复。

Abstract:

Objective To investigate the mechanism of interlekin 6 (IL 6) in wound healing of human biliary epithelial cells (BECs). Methods BECs were cultured in IL 6 at different concentrations: 0 ng/L (0 ng/L group), 10 ng/L (10 ng/L group), 50 ng/L (50 ng/L group), 100 ng/L (100 ng/L group), 1000 ng/L (1000 ng/L group). The effects of IL 6 on the phosphorylation of signal transducer and activator of transcription 3 (STAT3) and the expression of trefoil family factors 3 (TFF3) were detected. BECs were divided into untreated group, STAT3 RNAi group (BECs transfected with STAT3 RNAi adenovirus) and Control RNAi group (BECs transfected with vacant RNAi adenovirus). The effects of IL 6 on the expression of TFF3 were detected after RNAi of STAT3. In vitro wound models were constructed for the untreated group, STAT3 RNAi group and Control RNAi group, and the effects of IL 6 and TFF3 on BECs of the 3 groups were detected. All data were analyzed by using the Student′s t test, analysis of variance or Sidak test. Results The expressions of phosphorylated STAT3 in the 50 ng/L group, 100 ng/L group and 1000 ng/L group were 0.240±0.052, 0.714±0.124, 0.327±0.069, respectively, which were significantly higher than 0.033±0.011 of the 0 ng/L group (q=5.246, 17.260, 7.451, P<0.05). The contents of mRNA and protein of TFF3 increased as the increase of IL 6 concentration (q=12.045, 9.889, P<0.05). After RNAi of STAT3 of the BECs, the expression of TFF3 decreased when the concentration of IL 6 was 1000 ng/L. The expression of TFF3 of the STAT3 RNAi group was 0.037±0.005, which was significantly lower than 0.267±0.038 of the Control RNAi group and 0.301±0.042 in the untreated group (q=12.135, 13.929, P<0.05). In the in vitro wound model, the speed of BECs migration in the STAT3RNAi group was (9.1±1.5)μm/h, which was slower than (25.1±3.8)μm/h of the Control RNAi group after 12 hours of interference with IL 6 (q=7.737, P<0.05). The speed of BECs migration of STAT3 RNAi group was (39.2±4.7)μm/h after adding 1 g/L of recombinant TFF, which was significantly faster than that of the Control RNAi group (q=14.507, P<0.05). Conclusion IL 6 promotes cell migration and wound healing by activating STAT3 and up regulating TFF3 expression.

IL-6在人胆管上皮细胞损伤修复中的作用

Effects of interleukin 6 in wound healing of human biliary epithelial cells