中华消化外科杂志
中華消化外科雜誌
중화소화외과잡지
CHINESE JOURNAL OF DIGESTIVE SURGERY
2012年
5期
471-475
,共5页
蒋桂星%崔云甫%曹利平%邰升%钟翔宇%王志东
蔣桂星%崔雲甫%曹利平%邰升%鐘翔宇%王誌東
장계성%최운보%조리평%태승%종상우%왕지동
胆管上皮细胞%损伤修复%IL-6%信号转导子和转录激活子3%三叶因子3
膽管上皮細胞%損傷脩複%IL-6%信號轉導子和轉錄激活子3%三葉因子3
담관상피세포%손상수복%IL-6%신호전도자화전록격활자3%삼협인자3
Biliary epithelial cells%Wound healing%Interleukin-6%Signal transducer and activator of transcription 3%Trefoil family factor 3
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:0 ng/L组,10 ng/L组,50 ng/L组,100 ng/L组,1000 ng/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STAT3 RNAi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's t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50 ng/L组、100 ng/L组、1000 ng/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0 ng/L组的0.033±0.011(q=5.246,17.260,7.451,P＜0.05).TFF3 mRNA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1000 ng/L时,TFF3 mRNA及其蛋白表达有所回落.行干扰后,STAT3-RNAi组BECs TFF3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BECs 12 h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TFF31 g/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.
目的 探討IL-6在人膽管上皮細胞(BECs)損傷脩複中的作用機製.方法 IL-6按濃度分為5組:0 ng/L組,10 ng/L組,50 ng/L組,100 ng/L組,1000 ng/L組,將不同濃度的IL-6分彆榦預體外培養的BECs,併檢測IL-6對轉錄激活子3(STAT3)燐痠化和三葉因子3(TFF3)錶達的影響；將BECs分為未處理組、STAT3-RNAi組(細胞中轉入STAT3 RNAi腺病毒)和Control-RNAi組(細胞中轉入空RNAi腺病毒),檢測行榦擾後,IL-6對TFF3錶達的影響；分彆用未處理組、STAT3-RNAi組、Control-RNAi組BECs製備體外損傷模型,觀察IL-6、TFF3對各組細胞的影響.兩組數據間比較採用Student's t檢驗,多組間比較採用單因素方差或Sidak檢驗.結果 添加IL-6的50 ng/L組、100 ng/L組、1000 ng/L組燐痠化STAT3( p-STAT3)的錶達水平分彆為0.240±0.052、0.714 ±0.124、0.327± 0.069,明顯彊于0 ng/L組的0.033±0.011(q=5.246,17.260,7.451,P＜0.05).TFF3 mRNA及其蛋白錶達水平隨著IL-6濃度的增高而顯著增加(q=12.045,9.889,P ＜0.05)；IL-6濃度為1000 ng/L時,TFF3 mRNA及其蛋白錶達有所迴落.行榦擾後,STAT3-RNAi組BECs TFF3蛋白錶達水平為0.037±0.005,明顯低于Control-RNAi組的0.267±0.038和未處理組的0.301 ±0.042(q=12.135,13.929,P＜0.05).體外損傷模型中,IL-6榦預BECs 12 h後,STAT3-RNAi組BECs移行速度為(9.1±1.5)μm/h,慢于Control-RNAi組的(25.1±3.8)μm/h(q=7.737,P＜0.05)；而STAT3-RNAi組中加入外源性人重組TFF31 g/L後,BECs移行速度為(39.2±4.7)μm/h,比Control-RNAi組顯著提高(q=14.507,P＜0.05).結論 IL-6主要通過激活STAT3,繼髮上調TFF3錶達,從而促進人膽管上皮細胞移行和損傷脩複.
목적 탐토IL-6재인담관상피세포(BECs)손상수복중적작용궤제.방법 IL-6안농도분위5조:0 ng/L조,10 ng/L조,50 ng/L조,100 ng/L조,1000 ng/L조,장불동농도적IL-6분별간예체외배양적BECs,병검측IL-6대전록격활자3(STAT3)린산화화삼협인자3(TFF3)표체적영향；장BECs분위미처리조、STAT3-RNAi조(세포중전입STAT3 RNAi선병독)화Control-RNAi조(세포중전입공RNAi선병독),검측행간우후,IL-6대TFF3표체적영향；분별용미처리조、STAT3-RNAi조、Control-RNAi조BECs제비체외손상모형,관찰IL-6、TFF3대각조세포적영향.량조수거간비교채용Student's t검험,다조간비교채용단인소방차혹Sidak검험.결과 첨가IL-6적50 ng/L조、100 ng/L조、1000 ng/L조린산화STAT3( p-STAT3)적표체수평분별위0.240±0.052、0.714 ±0.124、0.327± 0.069,명현강우0 ng/L조적0.033±0.011(q=5.246,17.260,7.451,P＜0.05).TFF3 mRNA급기단백표체수평수착IL-6농도적증고이현저증가(q=12.045,9.889,P ＜0.05)；IL-6농도위1000 ng/L시,TFF3 mRNA급기단백표체유소회락.행간우후,STAT3-RNAi조BECs TFF3단백표체수평위0.037±0.005,명현저우Control-RNAi조적0.267±0.038화미처리조적0.301 ±0.042(q=12.135,13.929,P＜0.05).체외손상모형중,IL-6간예BECs 12 h후,STAT3-RNAi조BECs이행속도위(9.1±1.5)μm/h,만우Control-RNAi조적(25.1±3.8)μm/h(q=7.737,P＜0.05)；이STAT3-RNAi조중가입외원성인중조TFF31 g/L후,BECs이행속도위(39.2±4.7)μm/h,비Control-RNAi조현저제고(q=14.507,P＜0.05).결론 IL-6주요통과격활STAT3,계발상조TFF3표체,종이촉진인담관상피세포이행화손상수복.
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 STAT3-RNAi 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.
