聚肌苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立*

doi:10.3969/j.issn.1672-5069.2014.06.015

实用肝脏病杂志 ›› 2014, Vol. 17 ›› Issue (6): 619-623.doi: 10.3969/j.issn.1672-5069.2014.06.015

聚肌苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立^*

郭晓霞, 李良学, 武玉鹏, 贾丽莉, 刘改萍, 白崇智

030015太原市山西省中医院肝病科（郭晓霞）; 山西省中医药研究院中心实验室（武玉鹏,贾丽莉,白崇智）; 山西省中医院病理科（刘改萍）; 山西省中医药研究院硕士研究生（李良学）

收稿日期:2014-06-20 出版日期:2014-12-31 发布日期:2016-04-11
作者简介:郭晓霞,女,38岁,医学博士,副主任医师,硕士研究生导师。主要从事中医药防治慢性肝病研究。E-mail: gxxsunny@hotmail.com
基金资助:
山西省卫生厅科技攻关项目（编号：201301019）

Establishment of a mouse model of early primary biliary cirrhosis by polyinosinic-polycytidylic acid

Guo Xiaoxia, Li Liangxue, Wu Yupeng

Department of Liver Diseases,Traditional Chinese Medicine Hospital,Taiyuan 030012, Shanxi Province,China

Received:2014-06-20 Online:2014-12-31 Published:2016-04-11

摘要/Abstract

摘要： 目的探讨应用聚肌胞苷酸建立早期原发性胆汁性肝硬化动物模型。方法30只雌性C57BL/6小鼠被随机分为模型组和对照组。模型组小鼠腹腔注射聚肌胞苷酸（5 mg.kg^-1）,对照组小鼠注射等体积生理盐水, 均2次/w,于8 w、16 w和24 w分别检测血清抗线粒体抗体M2亚型、外周血CD4⁺CD25⁺Foxp³⁺计数,以及肝组织CK19表达情况。结果在24 w末,模型组AMA-M2阳性率为100%（5/5）,显著高于对照组（0/5）;在8 w、16 w和24 w,模型组小鼠外周血CD4⁺CD25⁺Foxp³⁺细胞计数分别为（3.48±0.95）%、（3.30±1.55）%和（2.67±0.97）%,均显著低于相应时间点对照组水平[（7.25±1.63）%、（6.33±1.06）%和（5.58±1.52）%,P<0.05];两组肝组织CK19表达无显著性差异;病理学检查显示模型组小鼠肝组织汇管区和胆管周围炎性细胞浸润呈进行性加重,细小胆管增生,出现胆汁性碎屑样坏死和桥接纤维化,但未出现肝硬化。结论聚肌胞苷酸注射可成功建立早期原发性胆汁性肝硬化动物模型,CD4⁺CD25⁺Foxp³⁺T细胞可能在发病中发挥重要作用。

关键词: 原发性胆汁性肝硬化, 聚肌胞苷酸, 动物模型, 小鼠

Abstract: Objective To establish an animal model of early primary biliary cirrhosis （PBC） by injection of polyinosinic-polycytidylic acid（Poly I:C）. Methods Thirty female C57BL/6 mice were randomly divided into model and control group. Mice in model group were injected with poly I:C at a dose of 5 mg.kg^-1 while animals in control with equal volume of saline for 24 weeks. The antimitochondrial antibody-M2（AMA-M2）,CD4⁺CD25⁺Foxp³⁺ cells were detected at 8,16 and 24 weeks. Histological changes of liver samples were evaluated with HE, PASM,and masson trichrom staining and CK19 was detected by anti-CD19 antibody staining at each stage. Results The positive rate of AMA-M2 in model group at week 24 was 100%（5/5）,significantly higher than in the control group（0/5）;The percentages of CD4⁺CD25⁺Foxp³⁺ Tregs in the peripheral blood in model group at week 8,16 and 24 were（3.48±0.95）%,（3.30±1.55）% and（2.67±0.97）%,all of which significantly lower than those in the control group at each corresponding time[（7.25±1.63）%,（6.33±1.06）% and（5.58±1.52）%,P<0.05];The expression of CK19 in liver tissues showed no significant difference between the two groups;Liver pathology showed progressively increased infiltration of inflammatory cells around the portal area and bile duct,small bile duct proliferation,bilious fragmental necrosis and bridging fibers in model group. Conclusions The animal model of early PBC is successfully established by injection of Poly I:C and the CD4⁺CD25⁺Foxp3⁺ T cells may play a critical role in the pathogenesis of the entity.

Key words: Primary biliary cirrhosis, Polyinosinic-polycytidylic acid, Mouse, Animal model

郭晓霞, 李良学, 武玉鹏, 贾丽莉, 刘改萍, 白崇智. 聚肌苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立^*[J]. 实用肝脏病杂志, 2014, 17(6): 619-623.

Guo Xiaoxia, Li Liangxue, Wu Yupeng. Establishment of a mouse model of early primary biliary cirrhosis by polyinosinic-polycytidylic acid[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2014, 17(6): 619-623.

参考文献

[1] Talwalkar JA,Souto E,Jorgensen RA,et al. Natural history of pruritus in primary biliary cirrhosis. Clin Gastroenterol Hepatol, 2003,1（4）:297-302.
[2] Hirschfield GM. Diagnosis of primary biliary cirrhosis. Best Pract Res Clin Gastroenterol,2011,25（6）:701-712.
[3] 段维佳,贾继东.原发性胆汁性肝硬化的诊断和治疗.胃肠病学,2009,14（4）:209-213.
[4] 马欢,王邦茂. 原发性胆汁性肝硬化动物模型研究进展. 国际消化病杂志,2012,32（2）:93-95.
[5] Concepcion AR,Medina JF. Approaches to the pathogenesis of primary biliary cirrhosis through animal models. Clinics Res Hepatol Gastroenterol,2012,36（1）:21-28.
[6] 许良中,杨文涛.免疫组织化学反应结果的判断标准.中国癌症杂志,1996,6（4）:229-231.
[7] Soslow RA,Dannenberg AJ,Rush D,et al. Cox-2 is expressed in human pulmonary,colonic,and mammary tumors. Cancer,2000,89（12）:2637-2645.
[8] Lleo A,Invernizzi P,Mackay IR,et al. Etiopathogenesis of primary biliary cirrhosis. World J Gastroenterol,2008,14（21）:3328-3337.
[9] McNally RJ,James PW,Ducker S,et al. Seasonal variation in the patient diagnosis of primary biliary cirrhosis:further evidence for an environmental component to etiology. Hepatology,2011,54（6）:2099-2103.
[10] Invernizzi P,Selmi C,Gershwin ME. Update on primary biliary cirrhosis. Dig Liver Dis,2010,42（6）:401-408.
[11] 刘然,陆伦根. 抗线粒体抗体对原发性胆汁性肝硬化诊断价值研究进展. 实用肝脏病杂志,2014,17（2）:218-221.
[12] Braun D,Geraldes P,Demengeot J. Type I interferon controls the onset and severity of 232232autoimmune manifestations in lpr mice. J Autoimmun,2003,20（1）:15-25.
[13] 自身免疫性肝病诊断和治疗指南.中华医学会风湿病学分会.中华风湿病学杂志,2011,15（8）:556-558.
[14] Hsia CC,Evarts RP,Nakatsukasa H,et al. Occurrence of oval-type cells in hepatitis B virus-associated human hepatocarcinogenesis. Hepatology,1992,16（6）:1327-1333.
[15] Kaplan MM,Gershwin ME. Primary biliary cirrhosis. N Engl J Med,2005,353（12）:1261-1273.
[16] Bernuzzi F,Fenoglio D,Battaglia F,et al. Phenotypical and functional alterations of CD8 regulatory T cells in primary biliary cirrhosis. J Autoimmun,2010,35（3）:176-180.
[17] 刘娟,曹雪涛. 2012年度免疫学研究重要进展. 中国免疫学杂志,2013,29（1）:3-13.
[18] Ramsdell F. Foxp3 and natural regulatory T cells:key to a cell lineage. Immunity,2003,19（2）:165-168.
[19] 许茵,杨小娟,蒯守刚. 原发性胆汁性肝硬化患者外周血CD4 + CD25 + Foxp 3+ 调节性T细胞检测及其临床意义. 中国实验诊断学,2014,18（4）:592-594.
[20] Muratori L,Longhi MS. The interplay between regulatory and effector T cells in autoimmune hepatitis:Implications for innovative treatment strategies. J Autoimmun,2013,46（17）:74-80.

聚肌苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立^*

Establishment of a mouse model of early primary biliary cirrhosis by polyinosinic-polycytidylic acid

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