Abstract
Background: Alcoholic liver disease (ALD) is one of the most common chronic liver diseases worldwide. However, the potential molecular mechanism in ALD development remains unclear. The objective of this work was to identify key molecules and demonstrate the underlying regulatory mechanisms.Methods: RNA-seq datasets were obtained from Gene Expression Omnibus (GEO), and key molecules in ALD development were identified with bioinformatics analysis. Alcoholic liver disease mouse and cell models were constructed using Lieber-DeCarli diets and alcohol medium, respectively. Quantitative real-time PCR and Western blotting were conducted to confirm the differential expression level. Dual-luciferase reporter assays were performed to explore the targeting regulatory relationship. Overexpression and knockdown experiments were applied to reveal the potential molecular mechanism in ALD development.Results: Between ALD patients and healthy controls, a total of 416 genes and 21 microRNAs (miRNAs) with significantly differential expression were screened. A comprehensive miRNA-mRNA network was established; within this network, the miR-182-5p/FOXO1 axis was considered a significant pathway in ALD lipid metabolism. Mouse and cell experiments validated that miR-182-5p was substantially higher in ALD than in normal livers, whereas the expression of FOXO1 was dramatically decreased by alcohol consumption (P < 0.05). Next, dual-luciferase reporter assays demonstrated that miR-182-5p directly targets the binding site of the FOXO1 3′UTR and inhibits its mRNA and protein expression. In addition, miR-182-5p was found to promote hepatic lipid accumulation via targeting the FOXO1 signaling pathway, and inhibition of the miR-182-5p/FOXO1 axis improved hepatic triglyceride (TG) deposition in ALD by regulating downstream genes involved in lipid metabolism.Conclusion: In summary, key molecules were identified in ALD development and a comprehensive miRNA–mRNA network was established. Meanwhile, our results suggested that miR-182-5p significantly increases lipid accumulation in ALD by targeting FOXO1, thereby providing novel scientific insights and potential therapeutic targets for ALD.
Highlights
Alcoholic liver disease (ALD), a chronic consequence of excessive long-term alcohol consumption, is among the most common chronic liver diseases
Gene ontology enrichment analysis (Figure 2F) further illustrated that common differentially expressed genes (DEGs) were highly involved in signal transduction, oxidation–reduction process, and extracellular matrix organization; Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the PI3K–Akt signaling pathway, focal adhesions, and ECM–receptor interaction were significantly associated with ALD development (Figure 2G)
Its inhibition had opposite effects on ALD. These results demonstrated that the miR-182-5p/FOXO1 axis participates in lipid metabolism in alcoholic liver by regulating the expression of key lipid biosynthesis and decompositionrelated genes (Figure 9)
Summary
Alcoholic liver disease (ALD), a chronic consequence of excessive long-term alcohol consumption, is among the most common chronic liver diseases. Alcohol directly stimulates the activation of innate immune cells and the secretion of inflammatory cytokines [4]. These pathologies are hallmarks of early-stage ALD, including alcoholic fatty liver disease (AFLD) and alcoholic hepatitis (AH); evidence has demonstrated that a lack of early and proper treatment for patients with these diseases markedly increases the risk of developing advanced stage ALD (liver fibrosis) and even alcoholic liver cirrhosis [5]. Alcoholic liver disease (ALD) is one of the most common chronic liver diseases worldwide. The objective of this work was to identify key molecules and demonstrate the underlying regulatory mechanisms
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
