Combination of CCl₄ with alcoholic and metabolic injuries mimics human liver fibrosis

Abstract

Metabolic and alcoholic liver injuries result in nonalcoholic (NAFLD) or alcoholic (ALD) fatty liver disease, respectively. In particular, presence of fibrosis in NAFLD and ALD requires treatment, but development of drugs is hampered by the lack of suitable models with significant fibrosis. The carbon tetrachloride (CCl4) liver fibrosis model does not reflect human NAFLD or ALD, but CCl4 may serve as a fibrosis accelerator in addition to another injury. Ethanol in drinking water (16%) or Western diet (WD) were administered for 7 wk in mice either alone or in combination with CCl4 intoxications. Extent of fibrosis, steatosis, and inflammation was assessed by histology, transcription, and biochemistry. Furthermore, transcription of fibrosis, proliferation, and inflammation-related genes was studied on human liver samples with fibrosis resulting from hepatitis C virus infection ( n = 7), NAFLD ( n = 8), or ALD ( n = 7). WD or ethanol alone induced only mild steatosis and inflammation. Combination of CCl4 and WD induced the most severe steatosis together with significant liver fibrosis and moderate inflammation. Combination of CCl4 and ethanol induced the strongest inflammation, with significant liver fibrosis and moderate steatosis. The relationship pattern between fibrosis, proliferation, and inflammation of human ALD was mostly similar in mice treated with CCl4 and ethanol. The combination of CCl4 intoxication with WD validates previous data suggesting it as an appropriate model for human nonalcoholic steatohepatitis. Especially, CCl4 plus ethanol for 7 wk induces ALD in mice, providing a model suitable for further basic research and drug testing. NEW & NOTEWORTHY Alcoholic fatty liver disease with significant fibrosis is generated within 7 wk using carbon tetrachloride as a fibrosis accelerator and administering gradually ethanol (up to 16%) in mice. The similarity in the pattern of steatosis, inflammation, and fibrosis involved in alcoholic fatty liver disease to those of the human condition renders this mouse model suitable as a preclinical model for drug development.

Systemic MCP-1 Levels Derive Mainly From Injured Liver and Are Associated With Complications in Cirrhosis

“…As previously described and published (12), male wild-type (WT, C57Bl6/J) mice (12 weeks old) were purchased (Charles River Laboratories Research Model and Services Germany, Sulzfeld, Germany). A total of 11 mice were used for this study.…”

1H-NMR Metabolomics Analysis of The Intervention Effects of Ruangan Xiaoji Decoction on CCl4 Induced Liver Fibrosis in Rats

“…The formation of free radicals and the chain peroxidation reaction caused by CCl 4 are the main mechanism of liver damage caused by CCl 4 . Liver cells are attacked by free radicals, and the permeability of the cell membrane increases, leading to the release of intracellular enzymes and increasing serum enzymes [18]. This experiment replicated the acute liver injury model of SD rats induced by CCl 4 .…”

Recent Advances in Practical Methods for Liver Cell Biology: A Short Overview

“…Alcoholic liver disease [6,7,[31][32][33][34] NASH/HCC (WD + CCl 4 ) Knock-out mice HFD HF-HC MCD Non-alcoholic liver disease [1,7,9,[35][36][37][38][39][40][41][42][43][44] Xenograft Isograft DEN DEN + CCL 4 NASH/HCC (WD + CCl 4 ) Hepatocarcinoma [7,9,[45][46][47][48][49] Hemodynamic assays Monitoring protocols HVPG CT-or MRI-based cross imaging Portal hypertension [39,[50][51][52] Knock-out mice, retroviral, lentiviral, sh/siRNA knock-down, CRISPR/Cas9-based method Angiogenesis [53][54][55][56] A-DW: alcohol in drinking water; APAP: acetaminophen; HF-HC: high fat/high cholesterol; HFD: high-fat diet; LPS: lipopolysaccharide; MACS: magnetic-activated cell sorting; MCD: methionine choline deficient; NIAAA: mouse model of chronic and binge ethanol feeding.…”

“…Using in vitro models of primary human liver cells from different human donors, co-culture systems can simulate additional pathophysiological liver cell interactions, being complementary to animal models in preclinical analysis. Furthermore, development of animal models to experimentally mimic alcoholic liver disease (ALD), such as the liquid Lieber-DeCarli (LDC) diet, intragastric ethanol infusion and administration of carbon tetrachloride (CCl 4 ) combined with ethanol in drinking water, can help to understand critical pathophysiological steps of human ALD, such as inflammation and fibrosis during ALD progression [6,7]. Moreover, HCC models are receiving increased attention, not only for the fast xenograft model, but also for application of N-nitrosodiethylamine (DEN) followed by repeated administration of CCl 4 (Table 1) [8], as well as the model of non-alcoholic steatohepatitis (NASH) combined with a HCC model described recently by using a Western diet (WD) combined with CCl 4 (Table 1) [7,9].…”