A high-fat diet and multiple administration of carbon tetrachloride induces liver injury and pathological features associated with non-alcoholic steatohepatitis in mice
A high-fat diet and multiple administration of carbon tetrachloride induces liver injury and pathological features associated with non-alcoholic steatohepatitis in mice
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DOI：
10.1111/1440-1681.12102
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发表时间：
2013-07-01
影响因子：
2.9
通讯作者：
Ishikawa, Fumiyasu
中科院分区：
文献类型：
--
作者：
Kubota, Norihiro;Kado, Shoichi;Ishikawa, Fumiyasu
本研究的目的是建立进行性脂肪性肝炎小鼠模型，因为很少有报道的非酒精性脂肪性肝炎（NASH）动物模型显示从脂肪肝到脂肪性肝炎的进展。C57 BL/6 N小鼠喂食高脂饮食（HFD）以产生肥胖，并给予四氯化碳（CCl 4）八次（0.05mL/kg，皮下注射，一次，随后0.1 mL/kg，s.c.，7次）或不。检测血清学指标和肝组织病理学检查。在一项单独的实验中，向HFD喂养的肥胖小鼠皮下注射0至8次CCl 4，以研究进行性变化。分析了氧化应激、炎症和细胞凋亡以及肝脏组织病理学变化的标志物。HFD喂养的肥胖小鼠显示脂肪肝，但没有脂肪性肝炎。相比之下，HFD喂养的小鼠给予CCl 4 8次，表现出脂肪性肝炎的组织病理学特征（脂肪肝、炎症、肝细胞气球样变和纤维化）和血清丙氨酸氨基转移酶水平升高。然而，向肥胖小鼠多次给予四氯化碳降低了还原型谷胱甘肽与氧化型谷胱甘肽的比率、超氧化物歧化酶活性和线粒体DNA拷贝数，导致慢性氧化应激的发展、凋亡细胞数量增加以及肿瘤坏死因子和转化生长因子mRNA水平升高。由此产生的炎症导致肝脏中羟脯氨酸含量增加和纤维化。本研究表明，多次给予HFD喂养的肥胖小鼠CCl 4可诱导慢性氧化应激，引发炎症和细胞凋亡，并导致肝脏纤维化的发展，导致脂肪肝进展为脂肪性肝炎。该模型可用于肝脏疾病的研究。
The aim of the present study was to establish a progressive steatohepatitis mouse model because few reported animal models of non-alcoholic steatohepatitis (NASH) show the progression from fatty liver to steatohepatitis. C57BL/6N mice were fed a high-fat diet (HFD) to develop obesity and were either administered carbon tetrachloride (CCl4) eight times (0.05mL/kg, s.c., once, followed by 0.1mL/kg, s.c., seven times) or not. Serum parameters and hepatic histopathology were examined. In a separate experiment, CCl4 was administered subcutaneously from 0 to eight times to HFD-fed obese mice to investigate progressive changes. Markers of oxidative stress, inflammation and apoptosis, as well as histopathological changes in the liver, were analysed. The HFD-fed obese mice showed fatty liver but not steatohepatitis. In contrast, HFD-fed mice administered CCl4 eight times showed histopathological features of steatohepatitis (fatty liver, inflammation, hepatocellular ballooning and fibrosis) and increased serum alanine aminotransferase levels. However, the multiple administration of CCl4 to obese mice reduced the ratio of reduced glutathione to oxidized glutathione, superoxide dismutase activity and mitochondrial DNA copy number, leading to the development of chronic oxidative stress, increased numbers of apoptotic cells and increased levels of both tumour necrosis factor- and transforming growth factor- mRNA. The resulting inflammation led to increased hydroxyproline content in the liver and fibrosis. The present study demonstrates that multiple administration of CCl4 to HFD-fed obese mice induces chronic oxidative stress that triggers inflammation and apoptosis and leads to the development of fibrosis in the liver, resulting in progression from fatty liver to steatohepatitis. This murine model will be useful in the research of hepatic disorders.