Ethanol with thioacetamide murine model of alcoholic liver disease identifies hepatic pathways as targets for the human disease

Ashi Mittal; Nishu Choudhary; Sudrishti Chaudhary; Anupama Kumari; Archana Rastogi; Guresh Kumar; Jaswinder Singh Maras; Shiv K Sarin; Shvetank Sharma

doi:10.1016/j.aohep.2024.101565

Ethanol with thioacetamide murine model of alcoholic liver disease identifies hepatic pathways as targets for the human disease

Ann Hepatol. 2024 Sep 12:101565. doi: 10.1016/j.aohep.2024.101565. Online ahead of print.

Authors

Ashi Mittal¹, Nishu Choudhary¹, Sudrishti Chaudhary¹, Anupama Kumari¹, Archana Rastogi², Guresh Kumar³, Jaswinder Singh Maras¹, Shiv K Sarin⁴, Shvetank Sharma⁵

Affiliations

¹ Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences, New Delhi, India.
² Department of Histopathology, Institute of Liver and Biliary Sciences, New Delhi, India.
³ Department of Biostatistics, Institute of Liver and Biliary Sciences, New Delhi, India.
⁴ Department of Hepatology, Institute of Liver and Biliary Sciences, New Delhi, India.
⁵ Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences, New Delhi, India. Electronic address: shvetanks@gmail.com.

PMID: 39276982
DOI: 10.1016/j.aohep.2024.101565

Abstract

Introduction and objectives: Hepatic proteome and gut microbiota alterations are known in alcohol-associated hepatitis (AAH). Current animal models sparsely mimic human AAH. We aimed to develop an murine model that closely resembled human AAH.

Materials and methods: Male C57BL/6N mice were pair-fed control/incremental ethanol Lieber-DeCarli diets and thioacetamide (TAA) for 12-weeks to induce AAH. Hepatic proteome was analyzed using LC-MS/MS. Gut-bacteria was determined using 16s-rRNA sequencing.

Results: Mice exposed to EtOH+TAA displayed higher expression of liver triglycerides (1.5-fold, p=0.001), pro-inflammatory (IL6, 1.5-fold, p=0.002 and TNFα, 1.7-fold, p=0.01), fibrotic (TGF-β, 2.7-fold, p=0.01 and Col1α1, 2-fold, p=0.01) and oxidative markers (GSH and SOD (-1.5 fold, p=0.004 & 0.005 respectively)) as compared to EtOH alone. Histology of EtOH+TAA liver displayed pericellular liver fibrosis, increased steatosis, and neutrophil infiltration, which resembled human AAH. In the 12wk EtOH+TAA group, Desulfobacteria, Campylobacteria, and Patescibacteria increased by 2-fold (p=0.02). Pathway combined score (CS, log10) in EtOH+TAA treatment showed upregulated hepatic ethanol oxidation (CS=1.93), fatty acid biosynthesis (CS=2.48), necrosis (CS=1.59), collagen formation (CS=1.28) and hypoxia (CS=0.68)and downregulated fatty acid beta-oxidation (CS=2.37), PPAR signaling (CS=1.35) fatty acid degradation (CS=2.35), bile acid metabolism (CS=1.87), and oxidative phosphorylation (CS=1.50), as observed in human disease.

Conclusions: Using an ethanol-thioacetamide combination in mice results in a faster establishment of AAH with fibrosis than previously known models. Differential protein expression strongly correlates with pathways found altered in human AAH, thus making the model mimic human disease better than other known models., respectively. Thioacetamide (TAA) was administered to enhance liver fibrosis and mimic human AAH.

Keywords: Alcoholic associated hepatitis; Gut microbiome; Hepatic proteome; Liver fibrosis; Mice model.