非酒精性脂肪性肝纤维化小鼠模型的建立及炎症因子的表达

doi:10.3969/j.issn.1674-5817.2017.01.005

摘要/Abstract

摘要： 目的建立非酒精性脂肪性肝纤维化小鼠模型,观察其病变及炎症因子的表达情况。方法 20只雄性C57BL/6J小鼠随机分成对照组、非酒精性脂肪性肝纤维化模型组。对照组小鼠予以蛋氨酸-胆碱充足(MCS)饲料喂养,模型组小鼠予以蛋氨酸-胆碱缺乏(MCD)饲料喂养造模。于造模8周末处死小鼠。观察肝脏组织病理学变化,检测小鼠血清草氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、胆固醇(TC)、甘油三酯(TG)肿瘤坏死因子-〈(TNF-〈)、白细胞介素-6(IL-6)水平。结果造模8周末,模型组小鼠体质量显著下降。肝组织病理学观察显示严重肝脂肪变,肝细胞水肿,可见点灶状坏死及淋巴细胞浸润,局灶见界面肝炎,汇管区扩大,纤维组织增生。模型组小鼠肝脏炎症活动度得分和纤维化得分均显著高于对照组(P<0.01)。模型组小鼠血清ALT、AST、TNF-〈、IL-6与对照组比较显著升高,而TG和TC水平显著下降(P<0.01)。结论小鼠经MCD饲料喂养8周出现严重肝脂肪变、肝纤维化和炎症因子高表达,表明成功诱导小鼠非酒精性脂肪性肝纤维化模型。该方法造模简单,成模快,存活率高,适合用于非酒精性脂肪性肝纤维化发病机制和药物干预等方面的研究。

关键词: 非酒精性脂肪性肝纤维化, 蛋氨酸-胆碱缺乏(MCD), C57BL/6J小鼠, 炎症因子

Abstract: Objective To establish the non-alcoholic fatty liver fibrosis model in mice and observe the pathological changes and the expression of inflammatory factors. Methods Twenty male C57BL/6J mice were randomly divided into control group and model group. The mice in control group were fed with methionine and choline-supplement(MCS) diet, and the model group were fed with methionine choline deficiency (MCD) diet. The mice were weighed and sacrificed at after 8 weeks feeding. The histopathological changes in liver were observed. The level of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol (TC), triglyceride (TG), tumor necrosis factor -〈 (TNF-〈) and interleukin (IL-6 ) were detected. Results The body weight of mice in the model group decreased significantly at the end of the study. Severe hepatic steatosis, liver cell edema, visible focal necrosis and focal lymphocytic infiltration, interface hepatitis, periportal enlargement, hyperplasia of fibrous tissue were observed.The scores of liver inflammation activity and fibrosis score in model group were significantly higher than those in control group (P<0.01).The level of serum ALT、AST、TNF-〈、IL-6 were significantly higher in the model group than that of control group (P<0.01). Conclusion After feeding mice with MCD diet feeding for 8 weeks, non-alcoholic fatty liver fibrosis model of mice was successfully induced. The modeling method was simple, rapid, and with has high survival rate. It is suitable for the pathogenesis and drug intervention study of nonalcoholic fatty liver fibrosis.

Key words: Non-alcoholic fatty liver Fibrosis, Methionine choline deficiency(MCD), C57BL/6J mice, Inflammatory factors

中图分类号:

Q95-33

杨华, 赵亚娟, 欧强. 非酒精性脂肪性肝纤维化小鼠模型的建立及炎症因子的表达[J]. 实验动物与比较医学, 2017, 37(1): 20-24.

YANG Hua, ZHAO Ya-Juan, OU Qiang. Establishment of Nonalcoholic Fatty Liver Fibrosis Model and Expression of Inflammatory Factors in Mice[J]. Laboratory Animal and Comparative Medicine, 2017, 37(1): 20-24.

参考文献

[1] WGO(World Gastroenterolopy Organiastion). World Gastroenterolopy Organiastion Global Guidelines Nonalcoholic Fatty Liver Disease and Nonalcoholic Steatohepatitis[J]. J Clin Gastroenterol, 2014, 48(6):467-473.
[2] Angulo P.Long-term mortality in nonalcoholic fatty liver disease: is liver histology of any prognostic significance[J].Hepatology, 2010, 51(2):373-375.
[3] Mishina M, Sakimura K.Conditional gene targeting on the pure C57BL/6 genetic background[J]. Neurosci Res, 2007, 58(2):105-112.
[4] Duval C, Teixeira-Clerc F, Leblanc AF, et al.Chronic exposure to low doses of Dioxin promotes liver fibrosis development in the C57BL6/J diet-induced obesity mouse?model[J].Environ Health Perspect, 2016 . [Epub ahead of print]. DOI: 10.1289/EHP316.
[5] Chen YJ, Myracle AD, Wallig MA, et al.Dietary broccoli protects against fatty liver development but not against progression of liver cancer in mice pretreated with diethylnitrosamine[J]. J Funct Foods, 2016, 24(1):57-62
[6] Yun-Hee L, Sou Hyun K, Sang-Nam K,et al.Sex-specific metabolic interactions between liver and adipose tissue in MCD diet-induced non-alcoholic fatty liver disease[J].Oncotarget, 2016, 7(30):46959-46971.
[7] 王振, 杨涛, 李世朋, 等. 脂肪肝动物模型研究进展[J]. 实用器官移植电子杂志, 2016, 4(2):105-108.
[8] Engstler AJ, Aumiller T, Degen C, et al.Insulin resistance alters hepatic ethanol metabolism: studies in mice and children with non-alcoholic fatty liver disease[J]. Gut, 2016, 65(9):1564-1571.
[9] Bavia L, Cogliati B,Dettoni JB,et al.The complement component C5 promotes liver steatosis and inflammation in murine non-alcoholic liver disease model[J]. Immunol Lett , 2016, 177(1):53-61.
[10] Di Pasqua L,Berardo C,Rizzo V,et al.MCD diet-induced steatohepatitis is associated with alterations in asymmetric dimethylarginine (ADMA) and its transporters[J]. Mol Cell Biochem, 2016, 419(2):147-155.
[11] Yamada T, Obata A, Kashiwagi Y, et a1.Gd-EOB-DTPA-enhanced-MR imaging in the inflammation stage of nonalcoholic steatohepatitis (NASH) in mice[J]. Magnetic Resonance Imaging, 2016, 34(6):724-729.
[12] 吴睿, 陈征, 徐亚运, 等. 高脂饲料诱导的非酒精性脂肪肝大鼠模型建立及其血浆nesfatin-1的变化[J]. 卫生研究,2016, 5(3):452-457.
[13] Anstee QM, Goldin RD.Mouse models in non-alcoholic fatty liver disease and steatohepatitis research[J]. Int J Exp Pathol,2006, 87(1):1-16.
[14] Boga S, Koksal AR, Alkim H, et a1. Plasma pentraxin 3 differentiates nonalcoholic steatohepatitis (NASH) from Non-NASH[J]. Metab Syndr Relat Disord, 2015, 13(9):393-399.
[15] 齐慧慧, 陈岳祥. 非酒精性脂肪性肝病常用动物模型的研究进展[J]. 国际消化病杂志, 2012, 23(3):145-147.
[16] Caballero F, Fernandez A, Matias N,et al.Specific contribution of methionine andcholine in nutritional nonalcoholic steatohepatitis: impact on mitochondrial S-adenosyl-L-methionine and glutathione[J]. J Biol Chem, 2010, 285(24):18528-18536.
[17] Kirsch R,Clarkson V,Shephard EG,et al.Rodent nutritional model of non-alcoholic steatohepatitis: species, strain and sex difference studies[J]. J Gastroenterol Hepatol, 2003, 18(11):272-1282.
[18] Yasuka M, Yusuke N, Toshiaki F, et al.LUBAC formation is impaired in the livers of mice with MCD-dependent nonalcoholic steatohepatitis[J]. Mediators Inflamm, 2015,2015:125380.
[19] 刘晓琳, 明雅南, 茅益民. 非酒精性脂肪性肝病及其伴随疾病动物模型的建立[J]. 肝脏, 2014, 15(9):374-376.
[20] Marcolin E, Forgiarini L, Tieppo J, et al.Methione-and choline-deficient diet induces hepatic changes characteristic of non-alcoholic steatohepatitis[J]. Arquivos De Gastroenterologia, 2011, 48(1):72-79.