Xiao Cheng,Kang Quan,Luo Qing.Establishment and evaluations of cholestatic cirrhosis model induced by bile duct ligation[J].Journal of Clinical Pediatric Surgery,2020,19(01):74-80.[doi:10.3969/j.issn.1671-6353.2020.01.015]
胆总管结扎胆汁淤积性肝硬化动物模型的建立与评价
- Title:
- Establishment and evaluations of cholestatic cirrhosis model induced by bile duct ligation
- Keywords:
- Common Bile Duct; Ligation; Liver Cirrhosis; Biliary; Model; Animal
- 分类号:
- R726;R575.7;R575.2
- 摘要:
- 目的 探讨胆总管结扎手术致胆汁淤积性肝硬化小鼠模型的建立价值，并对模型进行评价。方法 将84只balb/c小鼠随机分为实验组（采用胆总管结扎术）和对照组（采用假手术），术后观察小鼠一般情况和肝胆大体结构；采用全自动生化分析仪检测术后1 d、3 d、5 d、7 d、14 d、21 d、28 d总胆红素（total bilirubin，TBIL）、直接胆红素（direct bilirubin，DBIL）、谷氨酸转氨酶（alanine aminotransferase，ALT）、天门冬氨酸氨基转移酶（aspartate aminotransferase，AST）；采用H&E染色评估肝脏组织形态学变化；采用masson染色和免疫组织化学检测术后28 d小鼠肝脏组织切片，评估两组胶原纤维增生和a-SMA、CK-19蛋白表达情况。结果 实验组与对照组小鼠相比，术后一般情况相对更差，胆汁淤积，胆囊、胆总管体积明显增大，各项血清生化指标明显增高（P<0.05）；HE染色可见肝小叶结构改变，胆管增生；术后28 d实验组小鼠masson染色可见胶原纤维形成，免疫组织化学染色提示a-SMA、CK-19表达量显著增多（P<0.05）。结论 经腹中线小切口行胆总管结扎手术可成功构建小鼠胆汁淤积性肝硬化模型。
- Abstract:
- Objective To explore the establishment and evaluations of cholestatic cirrhosis induced by bile duct ligation in mice.Methods A total of 84 balb/c mice were randomly divided into experimental group (bile duct ligation) and control group (sham operation).General status and hepaticobiliary structures were observed postoperatively.And serum levels of total bilirubin (TBIL),alanine transaminase (ALT),direct bilirubin (DBIL) and aspartate transaminase (AST) were detected by an automatic biochemistry analyzer at Day 1/3/5/7/14/21/28 post-operation.And the histological changes of hepatic tissue were assessed by hematoxylin & eosin (H&E) stain while the hepatic expressions of collagen fiber,a-SMA and CK-19 protein at Day 28 was assessed by Masson’s stain and immunohistochemistry for evaluating the hepatic expressions of collagen fiber,a-SMA and CK-19 protein at 28 day post-operation.Results Compared with control group,mice in experimental group had worse general status,cholestasis,gallbladder & common bile duct expansion and cholestasis.Serum biochemical parameters significantly increased (P<0.05).H&E stain showed changes in hepatic lobule and bile duct proliferation.Collagen fiber formation was observed by Masson stain in experimental group at Day 28 day post-operation and immunohistochemical staining indicated that a-SMA and CK-19 increased markedly. Conclusion A murine model of cholestatic cirrhosis is established successfully by bile duct ligation through small incision of abdominal midline.
参考文献/References:
1 Galicia-Moreno M,Favari L,Muriel P.Trolox mitigates fibrosis in a bile duct ligation model[J].Fundam Clin Pharmacol,2013,27(3):308-318.DOI:10.1111/j.1472-8206.2011.01020.x.
2 Bai H,Zhang N,Xu Y,et al.Yes-associated protein regulates the hepatic response after bile duct ligation[J].Hepatology,2012,56(3):1097-1107.DOI:10.1002/hep.25769.
3 Meng F,Wang K,Aoyama T,et al.Interleukin-17 signaling in inflammatory,Kupffer cells,and hepatic stellate cells exacerbates liver fibrosis in mice[J].Gastroenterology,2012,143(3):765-776.e3.DOI:10.1053/j.gastro.2012.05.049.
4 Goncalves JO,Tannuri AC,Coelho MC,et al.Dynamic expression of desmin,alpha-SMA and TGF-beta1 during hepatic fibrogenesis induced by selective bile duct ligation in young rats[J].Braz J Med Biol Res,2014,47(10):850-857.DOI:10.1590/1414-431x20143679.
5 Weerachayaphorn J,Luo Y,Mennone A,et al.Deleterious effect of oltipraz on extrahepatic cholestasis in bile duct-ligated mice[J].J Hepatol,2014,60(1):160-166.DOI:10.1016/j.jhep.2013.08.015.
6 Tag CG,Sauer-Lehnen S,Weiskirchen S,et al.Bile duct ligation in mice:induction of inflammatory liver injury and fibrosis by obstructive cholestasis[J].J Vis Exp,2015,(96).DOI:10.3791/52438.
7 Tag CG,Weiskirchen S,Hittatiya K,et al.Induction of experimental obstructive cholestasis in mice[J].Lab Anim,2015,49(1 Suppl):70-80.DOI:10.1177/0023677214567748.
8 Zhang Y,Hong JY,Rockwell CE,et al.Effect of bile duct ligation on bile acid composition in mouse serum and liver[J].Liver Int,2012,32(1):58-69.DOI:10.1111/j.1478-3231.2011.02662.x.
9 Heinrich S,Georgiev P,Weber A,et al.Partial bile duct ligation in mice:a novel model of acute cholestasis[J].Surgery,2011,149(3):445-451.DOI:10.1016/j.surg.2010.07.046.
10 Kirkland JG,Godfrey CB,Garrett R,et al.Reversible surgical model of biliary inflammation and obstructive jaundice in mice[J].J Surg Res,2010,164(2):221-227.DOI:10.1016/j.jss.2009.08.010.
11 肖颜玉,胥飚,崔鲂,等.小鼠胆总管结扎致肝纤维化模型的建立与评价[J].重庆医科大学学报,2008,33(4):390-393.DOI:10.13406/j.cnki.cyxb.2008.04.012. Xiao YY,Xu B,Cui F,et al.Establishment and evaluation of liver fibrosis model induced by bile duct ligation in mouse[J].Journal of Chongqing Medical University,2008,33(4):390-393.DOI:10.13406/j.cnki.cyxb.2008.04.012.
12 罗丹,文阳安,董玉芳,等.胆总管结扎引起胆汁淤积对小鼠肝脏细胞增殖的影响[J].第四军医大学学报,2009,30(2):134-137.DOI:10.3321/j.issn:1000-2790.2009.02.011. Luo D,Wen YA,Dong YF,et al.Effect of bile duct ligation induced biliary cholestasis on hepatocytes proliferation[J].Journal of Fourth Military Medical University,2009,30(2):134-137.DOI:10.3321/j.issn:1000-2790.2009.02.011.
13 Huss S,Schmitz J,Goltz D,et al.Development and evaluation of an open source Delphi-based software for morphometric quantification of liver fibrosis[J].Fibrogenesis Tissue Repair,2010,3(1):10.DOI:10.1186/1755-1536-3-10.
14 宣佶,田耀洲,曹鹏,等.胆汁淤积性肝硬化大鼠模型的改良[J].中国比较医学杂志,2014,24(4):57-61.DOI:10.3969/j.issn.1671.7856.2014.004.013. Xuan J,Tian YZ,Cao P,et al.Improvement in the rat model of liver fibrosis induced by bile duct ligation[J].Chinese Journal of Comparative Medicine,2014,(4):57-61.DOI:10.3969/j.issn.1671.7856.2014.004.013.
15 Yuan S,Jiang T,Zheng R,et al.Effect of bone marrow mesenchymal stem cell transplantation on acute hepatic failure in rats[J].Exp Ther Med,2014,8(4):1150-1158.DOI:10.3760/cma.j.issn.1007-3418.2018.10.004.
16 Gon?alves JO,Tannuri AC,Coelho MC,et al.Dynamic expression of desmin,α-SMA and TGF-β1 during hepatic fibrogenesis induced by selective bile duct ligation in young rats[J].Braz J Med Biol Res,2014,47(10):850-857.DOI:10.1590/1414-431x20143679.
17 Deng YH,Pu CL,Li YC,et al.Analysis of biliary epithelial-mesenchymal transition in portal tract fibrogenesis in biliary atresia[J].Dig Dis Sci,2011,56(3):731-740.DOI:10.1007/s10620-010-1347-6.
18 Gressner AM,Weiskirchen R.Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets[J].J Cell Mol Med,2006,10(1):76-99.DOI:10.1111/j.1582-4934.2006.tb00292.x.
19 Liu WH,Ren LN,Chen T,et al.Stages based molecular mechanisms for generating cholangiocytes from liver stem/progenitor cells[J].World J Gastroenterol,2013,19(41):7032-7041.DOI:10.3748/wjg.v19.i41.7032.
20 Zhou J,Zhao L,Qin L,et al.Epimorphin regulates bile duct formation via effects on mitosis orientation in rat liver epithelial stem-like cells[J].PLoS One,2010,5(3):e9732.DOI:10.1371/journal.pone.0009732.
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备注/Memo
收稿日期:2017-08-25。
基金项目:国家自然科学基金资助项目（编号：NSFC81172545）；重庆市科委基础与前沿研究计划资助项目（编号：cstc2013jcyA10026）
通讯作者:康权,Email:564799351@qq.com