摘要
目的观察胆总管结扎诱导的梗阻性黄疸大鼠成纤维细胞活化的影响。方法雄性无特定病原体(SPF)级大鼠(第二军医大学实验动物中心提供)24只随机分为假手术组与实验组,每组大鼠各12只。所有大鼠均进行胆总管结扎。实验组大鼠用3-0丝线双重结扎且于结间切断胆总管。两组大鼠分别于术后2 d和7 d取材。采用酶联免疫吸附试验(ELISA法)检测人转化生长因子-β1(TGF-β1)含量;采用免疫组织化学法检测TGF-β1蛋白表达;采用实时荧光定量聚合酶链反应(FQ-PCR)法检测TGF-β1 mRNA表达;采用蛋白质印迹法(Western blot)检测TGF-β1蛋白表达。应用SPSS 22.0统计软件分析,计量资料采用t检验。结果假手术组大鼠肝小叶结构清晰,肝索排列整齐;实验组大鼠肝小叶结构紊乱不清,并且肝索解离。实验组血清TGF-β1水平术后2 d[(689.42±58.13)ng/L]和术后7 d[(867.53±45.62)ng/L]高于假手术组[(519.92±32.41)ng/L和(536.52±41.29)ng/L,t=8.822、18.635,P<0.05],差异有统计学意义。实验组肝组织中TGF-β1蛋白表达术后2 d[(6.53±1.29)%]和术后7 d[(8.97±1.85)%]高于假手术组[(1.17±0.24)%和(1.54±0.39)%,t=14.151、13.613,P<0.05],差异有统计学意义。实验组肝组织中TGF-β1 mRNA表达术后2 d[(0.89±0.14)%]和术后7 d[(0.95±0.19)%]高于假手术组[(0.52±1.32)%和(0.31±1.76)%,t=14.693、18.316,P<0.05],差异有统计学意义。实验组肝组织中TGF-β1蛋白表达灰度值术后2 d(0.62±0.13)和术后7 d(0.78±0.10)高于假手术组(0.21±0.05和0.23±0.06,t=10.197、16.337,P<0.05),差异有统计学意义。结论胆总管结扎诱导的梗阻性黄疸大鼠TGF-β1表达明显上调,胆管上皮细胞增殖诱导成纤维细胞活化。
Objective To investigate the effect of common bile duct ligation on fibroblast activation in rats with obstructive jaundice.Methods Twenty-four male specific pathogen free(SPF)rats were randomly divided into sham-operated group and experimental group,with 12 rats in each group.All rats underwent common bile duct ligation.Rats The common bile duct in the experimental group was ligated with 3-0 silk thread and cut off.The rats in the two groups were sampled 2 and 7 days after operation.Enzyme-linked immunosorbent assay(ELISA)was used to detect the content of transforming growth factor-β1(TGF-β1).Immunohistochemistry was used to detect the expression of TGF-β1 protein.The real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)was used to detect the expression of TGF-β1 gene.Western blotting was used to detect the expression of TGF-β1 protein.Results In sham-operated group,the structure of hepatic lobules was intact,and the hepatic sinuses were normal without congestion.In experimental group,the structure of hepatic lobules was disordered and the hepatic cords were dissociated.The level of serum TGF-β1 in the experimental group at 2nd day[(689.42±58.13)ng/L]and 7th day[(867.53±45.62)ng/L]was significantly higher than that in the sham-operated group[(519.92±32.41)ng/L and(536.50±41.29)ng/L,t=8.822 and 18.635,P<0.05].The expression of TGF-β1 protein in liver tissues of experimental group at 2nd day[(6.53±1.29)%]and 7th day[(8.97±1.85)%]was significantly higher than that of sham-operated group[(1.17±0.24)%and(1.54±0.39)%,t=8.822 and 18.635,P<0.05].The expression of TGF-β1 in liver tissues of experimental group at 2nd day[(0.89±0.14)%]and 7th day[(0.95±0.19)%]was significantly higher than that of sham-operated group[(0.52±1.32)%and(0.31±1.76)%,t=14.693 and 18.316,P<0.05].The gray value of TGF-β1 protein expression in liver tissue of experimental group at 2nd day after operation(0.62±0.13)and 7th day after operation(0.78±0.10)was significantly higher than that of sham-operated group(0.21±0.05 and 0.23±0.06,t=10.197 and 16.337,P<0.05).Conclusion The expression of TGF-β1 in rats with obstructive jaundice induced by common bile duct ligation is significantly up-regulated,and the proliferation of biliary duct epithelial cells induces fibroblast activation.
作者
张清华
欧贵雪
王瑞华
余振兴
刘景丰
Zhang Qinghua;Ou Guixue;Wang Ruihua;Yu Zhenxing;Liu Jingfeng(Department of General Surgery,Mindong Hospital Affiliated to Fujian Medical University,Fu’an 355000,China;Department of Hepatobiliary Surgery,Mengchao Hepatobiliary Hospital of Fujian Medical University,Fuzhou 350025,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第1期60-62,共3页
Chinese Journal of Experimental Surgery
基金
福建省自然科学基金(2016J01660)。
关键词
胆总管结扎
梗阻性黄疸
成纤维活化
Common bile duct ligation
Obstructive jaundice
Fibroblast activation
