Adult mouse model of early hepatocellular carcinoma promoted by alcoholic liver disease
Adult mouse model of early hepatocellular carcinoma promoted by alcoholic liver disease
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DOI：
10.3748/wjg.v22.i16.4091
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发表时间：
2016-04-28
影响因子：
4.3
通讯作者：
Szabo, Gyongyi
中科院分区：
文献类型：
--
作者：
Ambade, Aditya;Satishchandran, Abhishek;Szabo, Gyongyi
目的：方法：成年C57 BL/6雄性小鼠，给予化学致癌物二乙基亚硝胺（DEN）多次给药，随后给予4% Lieber-DeCarli饲料喂养7 wk，建立酒精性肝细胞癌（HCC）小鼠模型。检测血清丙氨酸氨基转移酶（ALT）、甲胎蛋白（AFP）和肝脏Cyp 2 e1。检测巨噬细胞F4/80、CD 68和中性粒细胞Ly 6 G、MPO、E-选择素的表达。通过IL-1 β/iNOS（M1）和Arg-1/IL-10/CD 163/CD 206（M2）表达确定巨噬细胞极化。油红O染色法检测肝脂肪变性，天狼星红染色法检测肝纤维化。通过磁共振成像监测HCC的发展，并通过组织学证实。结果：酒精-DEN小鼠在整个酒精喂养过程中均表现出比配对喂养的DEN小鼠更高的ALT，而体重没有增加。与配对喂养的对照组相比，酒精喂养导致ALT、肝脏脂肪变性和炎症增加。酒精-DEN小鼠脂肪变性减少，纤维化增加，表明晚期肝病。分子特征显示，酒精DEN肝脏中的中性粒细胞和巨噬细胞标志物水平最高。重要的是，M2巨噬细胞在酒精DEN肝脏中主要较高。磁共振成像显示肝内囊肿数量增加，肝脏组织学证实酒精DEN小鼠与所有其他组相比存在早期HCC。这与酒精-DEN小鼠中血清甲胎蛋白（HCC的标志物）升高相关。PCNA免疫组化显示显着增加肝细胞增殖的肝脏酒精DEN相比，对喂食DEN或alcohol-fed mice.CONCLUSION：我们描述了一个新的12周肝癌模型在成年小鼠中，酒精性肝炎的肝脏发展，并定义ALD作为辅助因子在肝癌。
AIM: to establish a mouse model of alcohol-driven hepatocellular carcinoma (HCC) that develops in livers with alcoholic liver disease (ALD).METHODS: Adult C57BL/6 male mice received multiple doses of chemical carcinogen diethyl nitrosamine (DEN) followed by 7 wk of 4% Lieber-DeCarli diet. Serum alanine aminotransferase (ALT), alpha fetoprotein (AFP) and liver Cyp2e1 were assessed. Expression of F4/80, CD68 for macrophages and Ly6G, MPO, E-selectin for neutrophils was measured. Macrophage polarization was determined by IL-1 beta/iNOS (M1) and Arg-1/IL-10/CD163/CD206 (M2) expression. Liver steatosis and fibrosis were measured by oil-red-O and Sirius red staining respectively. HCC development was monitored by magnetic resonance imaging, confirmed by histology. Cellular proliferation was assessed by proliferating cell nuclear antigen (PCNA).RESULTS: Alcohol-DEN mice showed higher ALTs than pair fed-DEN mice throughout the alcohol feeding without weight gain. Alcohol feeding resulted in increased ALT, liver steatosis and inflammation compared to pair-fed controls. Alcohol-DEN mice had reduced steatosis and increased fibrosis indicating advanced liver disease. Molecular characterization showed highest levels of both neutrophil and macrophage markers in alcohol-DEN livers. Importantly, M2 macrophages were predominantly higher in alcohol-DEN livers. Magnetic resonance imaging revealed increased numbers of intrahepatic cysts and liver histology confirmed the presence of early HCC in alcohol-DEN mice compared to all other groups. This correlated with increased serum alpha-fetoprotein, a marker of HCC, in alcohol-DEN mice. PCNA immunostaining revealed significantly increased hepatocyte proliferation in livers from alcohol-DEN compared to pair fed-DEN or alcohol-fed mice.CONCLUSION: We describe a new 12-wk HCC model in adult mice that develops in livers with alcoholic hepatitis and defines ALD as co-factor in HCC.