Exendin-4 alleviates steatosis in an in vitro cell model by lowering FABP1 and FOXA1 expression via the Wnt/-catenin signaling pathway

Olfa Khalifa,Khaoula Errafii,Abdelilah Arredouani,Neyla S Al-Akl

doi:10.1038/s41598-022-06143-5

Olfa Khalifa, Khaoula Errafii + Show 2 more

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https://doi.org/10.1038/s41598-022-06143-5

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Journal: Scientific Reports	Publication Date: Feb 9, 2022
Citations: 22	License type: open-access

Affiliation: Hamad bin Khalifa University

Abstract

Non-alcoholic fatty liver disease (NAFLD) is the leading chronic liver disease worldwide. Agonists of the glucagon-like peptide-1 receptor (GLP-1R), currently approved to treat type 2 diabetes, hold promise to improve steatosis and even steatohepatitis. However, due to their pleiotropic effects, the mechanisms underlying their protective effect on NAFLD remain elusive. We aimed to investigate these mechanisms using an in vitro model of steatosis treated with the GLP-1R agonist Exendin-4 (Ex-4). We established steatotic HepG2 cells by incubating the cells with 400 µM oleic acid (OA) overnight. Further treatment with 200 nM Ex-4 for 3 h significantly reduced the OA-induced lipid accumulation (p < 0.05). Concomitantly, Ex-4 substantially reduced the expression levels of Fatty Acid-Binding Protein 1 (FABP1) and its primary activator, Forkhead box protein A1 (FOXA1). Interestingly, the silencing of β-catenin with siRNA abolished the effect of Ex-4 on these genes, suggesting dependency on the Wnt/β-catenin pathway. Additionally, after β-catenin silencing, OA treatment significantly increased the expression of nuclear transcription factors SREBP-1 and TCF4, whereas Ex-4 significantly decreased this upregulation. Our findings suggest that direct activation of GLP-1R by Ex-4 reduces OA-induced steatosis in HepG2 cells by reducing fatty acid uptake and transport via FABP1 downregulation.

Highlights

Non-alcoholic fatty liver disease (NAFLD), defined as the excessive accumulation of lipids in the liver, is the most common cause of chronic liver disease in industrialized nations[1] and the most frequent indication for liver transplantation[2,3]
By treating HepG2 cells with increasing Oleic acid (OA) concentrations for 16 h and measuring TG accumulation, we determined the optimal concentration of OA required to induce steatosis (Fig. 1A)
We have looked at the effect of Ex-4 on BODIPY staining in the absence of OA and found that it is significantly lower than OA alone (Fig. 1C,D)

Summary

Introduction

Non-alcoholic fatty liver disease (NAFLD), defined as the excessive accumulation of lipids in the liver, is the most common cause of chronic liver disease in industrialized nations[1] and the most frequent indication for liver transplantation[2,3]. Losing 5% of one’s bodyweight improves abnormal liver tests and reduces liver fat[9], whereas losing 7 to 10% of one’s body weight appears to reduce inflammation and injury to liver cells and may even reverse some fibrosis damage[10]. GLP1 regulates blood glucose levels by stimulating glucose-dependent insulin release and decreasing glucagon secretion, promotes proliferation of pancreatic b-cells, slows gastric emptying, Scientific Reports | (2022) 12:2226.

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