【摘要】：目的探討TOLL樣受體4(TLR4)在脂多糖(LPS)誘導的大鼠肝內膽管組織損傷中的作用及機制。方法將48只SD大鼠隨機均分為四組,LPS+siRNA轉染組和LPS+陰性病毒組經尾靜脈分別注入siRNA-TLR4純化腺病毒、陰性對照病毒,對照組和LPS組均經尾靜脈注射等量生理鹽水。給藥后24 h,LPS組、LPS+siRNA轉染組和LPS+陰性病毒組均經膽總管內緩慢注入LPS,對照組注入等量生理鹽水。制模后48、72 h各組分別取6只大鼠,采集肝門部包含膽管的肝組織標本。采用HE染色觀察肝門部肝內膽管組織炎性細胞浸潤情況及膽管形態(tài)等病理變化。采用免疫組化法檢測膽管上皮細胞TLR4、細胞角蛋白19(CK19)、波形蛋白(Vimentin)蛋白表達,采用RT-PCR法檢測膽管組織TLR4、CK19、Vimentin mRNA表達。結果 LPS組造模48、72 h時膽管周圍炎性細胞浸潤且膽管間質水腫,伴周圍纖維結締組織輕度增生;LPS+陰性病毒組表現(xiàn)與LPS組相似;LPS+siRNA轉染組造模48 h時膽管周圍僅有少量炎性細胞,造模72 h時膽管周圍炎性細胞稍增多,但較LPS組顯著減輕。與對照組比較,其他三組造模48、72 h時膽管上皮細胞CK19蛋白及mRNA表達均降低,TLR4、Vimentin蛋白及mRNA表達均升高,組間比較P均0.01;LPS+siRNA轉染組造模48、72 h時膽管上皮細胞CK19蛋白及mRNA表達均高于LPS組、LPS+陰性病毒組,TLR4、Vimentin蛋白及mRNA表達均低于LPS組、LPS+陰性病毒組,組間比較P均0.01。結論 LPS誘發(fā)肝內膽管組織發(fā)生炎癥反應及纖維化損傷過程中產生大量TLR4,TLR4可導致肝膽管上皮細胞發(fā)生上皮間質轉化,從而加重纖維化損傷程度。
[Abstract]:Aim to investigate the role and mechanism of TOLL-like receptor 4 (TLR4) in lipopolysaccharide (LPS)-induced intrahepatic bile duct injury in rats. Methods Forty-eight SD rats were randomly divided into four groups:, LPS siRNA transfection group and LPS negative virus group. SiRNA-TLR4 purified adenovirus was injected into the tail vein, and the negative control virus was injected into the tail vein respectively. The control group and the LPS group were injected with the same amount of normal saline through the tail vein. At 24 h after administration, LPS siRNA-transfected group and LPS-negative virus group were injected with the same amount of normal saline into the common bile duct by slow injection into the common bile duct of LPS, control group. At 48 h and 72 h after model, 6 rats in each group were taken and the liver tissue specimens containing bile duct in the hilum were collected. HE staining was used to observe the infiltration of inflammatory cells in the intrahepatic bile duct and the morphological changes of the bile duct. The expression of cytokeratin 19 (CK19) and vimentin (Vimentin) in bile duct epithelial cells (TLR4,) was detected by immunohistochemical method, and the expression of TLR4,CK19,Vimentin mRNA in bile duct tissue was detected by RT-PCR method. Results at 48 h and 72 h after LPS, inflammatory cells infiltrated around the bile duct and edema of the bile duct stroma, with slight proliferation of the surrounding fibrous connective tissue, and the expression of LPS-negative virus group was similar to that of the LPS group. There were only a few inflammatory cells around the bile duct in the LPS siRNA transfection group at 48 h, and a little more inflammatory cells in the bile duct around the bile duct at 72 h after the model was made, but decreased significantly than that in the LPS group. Compared with the control group, the expression of CK19 protein and mRNA in bile duct epithelial cells decreased and the expression of TLR4,Vimentin protein and mRNA increased in the other three groups (P < 0.01). The expression of CK19 protein and mRNA in bile duct epithelial cells of LPS siRNA transfection group was higher than that of LPS group, LPS negative virus group, TLR4,Vimentin protein and mRNA expression of LPS negative virus group were lower than that of LPS group, LPS negative virus group at 72 h after transfection, and the expression of TLR4,Vimentin protein and mRNA was significantly higher than that of LPS negative virus group (P < 0.01). Conclusion the inflammatory reaction of intrahepatic bile duct tissue induced by LPS and the production of a large number of TLR4,TLR4 in the process of fibrosis injury can lead to epithelial stroma transformation of hepatobiliary epithelial cells, thus aggravating the degree of fibrosis injury.
【作者單位】： 河南省省立醫(yī)院;遵義醫(yī)學院附屬醫(yī)院;
【基金】：國家自然科學基金資助項目(81260085)
【分類號】：R575.62

【相似文獻】

相關期刊論文 前9條

1 萬春,陳孝平;兔肝細胞膽管上皮細胞對射頻熱效應的敏感性[J];腹部外科;2005年02期

2 趙禮金;楊日高;程龍;王麥建;江艷;余德剛;周廷梅;王曙光;;肝內膽管上皮細胞上皮-間葉轉化的實驗研究[J];第三軍醫(yī)大學學報;2010年03期

3 朱列偉,俞健,曾紀驊,時毓民,陳蓮,周麗君;小膽管上皮細胞體積量化在實驗性黃疸模型中意義的探討[J];中國實驗動物學報;2000年02期

4 鄭兆齡;張啟發(fā);;肝機能不全[J];江西醫(yī)藥;1963年12期

5 李敏;田雨;吳碩東;李勇男;;脂多糖對肝內膽管上皮細胞MUC5AC表達的影響[J];世界華人消化雜志;2013年22期

6 王兵;劉平;龍愛華;陸雄;顧宏圖;胡義揚;劉成海;徐列明;;膽管上皮細胞增生在膽管結扎大鼠膽汁性肝纖維化形成中的作用[J];肝臟;2007年04期

7 鄔善敏,戴植本;表皮生長因子受體在梗阻性黃疸肝內膽管上皮細胞的表達及意義[J];中華實驗外科雜志;1997年04期

8 陳輝星;尹路;彭承宏;;人_β-2防御素在人胃腸膽管上皮細胞中的免疫作用[J];國際外科學雜志;2006年02期

9 ;[J];;年期

相關會議論文 前3條

1 王兵;劉平;;膽管上皮細胞與肝病[A];第十三次全國中西醫(yī)結合肝病學術會議論文匯編[C];2004年

2 方泰石;朱廣勇;邰升;;建立膽管上皮細胞H_2O_2損傷模型的心得體會[A];中國抗癌協(xié)會膽道腫瘤專業(yè)委員會成立大會暨第一屆全國膽道腫瘤學術會議論文集[C];2009年

3 楊再興;仲人前;;線粒體M2蛋白通過TLR4通路對人肝內膽管上皮細胞的作用研究[A];全國臨床免疫檢驗研討會暨第六屆全國臨床免疫學術會議論文匯編[C];2009年

相關碩士學位論文 前3條

1 韓瓏;脂多糖對人正常肝內膽管上皮細胞TLR4-NF-κB通路的影響[D];安徽醫(yī)科大學;2015年

2 王芳;LPS通過激活TLR4誘導人肝內膽管上皮細胞上皮—間質轉化[D];遵義醫(yī)學院;2016年

3 顧進;TLR4-snail信號通路在LPS誘導的人膽管上皮細胞EMT中作用初步研究[D];遵義醫(yī)學院;2015年

，

本文編號：2474452