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Updated: May 6, 2026
Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis
Published on: March 11, 2017
Association Between Active DNA Demethylation and Liver Fibrosis in Individuals with Metabolic-Associated Steatotic
Ilaria Barchetta1, Michele Zampieri1, Flavia Agata Cimini1
1Department of Experimental Medicine, Sapienza University of Rome, 00161 Rome, Italy.
Abstract:
Metabolic-associated steatotic liver disease (MASLD) represents the most common chronic hepatopathy worldwide and an independent risk factor for cardiovascular disease and mortality, particularly when liver fibrosis occurs. Epigenetic alterations, such as DNA methylation, may influence MASLD susceptibility and progression; yet mechanisms underlying this process are limited. This study aimed to investigate whether active DNA demethylation in peripheral blood mononuclear cells (PBMCs) from individuals with MASLD, alongside the methylation and mRNA levels of inflammation- and fibrosis-related candidate genes, is associated with liver fibrosis. For this study, global demethylation intermediates (5-hydroxymethylcytosine [5hmC], 5-formylcytosine [5fC]) were quantified in PBMCs from 89 individuals with/without MASLD using ELISA. Site-specific DNA methylation of SOCS3, SREBF1, and TXNIP was analyzed by mass spectrometry-based bisulfite sequencing; mRNA expression was assessed via RT-PCR. Individuals with MASLD and moderate-to-high fibrosis risk (estimated by the fibrosis non-alcoholic steatohepatitis (NASH) index, FNI) progressively exhibited greater global 5hmC and 5fC levels. Higher FNI was associated with reduced methylation of the SOCS3 gene and increased mRNA expression of the SOCS3, TXNIP, IL-6, and MCP-1 genes. In conclusion, elevated fibrosis risk in MASLD is associated with active global DNA demethylation, as well as differential methylation and expression patterns of genes, which are key regulators of inflammation and fibrosis. These epigenetic alterations in PBMCs may mirror DNA methylation changes in the liver, which may potentially contribute to liver fibrogenesis and represent novel biomarkers for MASLD progression toward fibrosis.
Insights
Metabolic-associated steatotic liver disease (MASLD) with fibrosis risk shows increased DNA demethylation in blood cells. Epigenetic changes in these cells may reflect liver alterations and serve as biomarkers for MASLD progression.
Area of Science:
- Hepatology
- Epigenetics
- Molecular Biology
Background:
- Metabolic-associated steatotic liver disease (MASLD) is a prevalent liver condition linked to cardiovascular risks, especially with fibrosis.
- Epigenetic modifications, including DNA methylation, are implicated in MASLD but underlying mechanisms remain unclear.
- Investigating epigenetic changes in peripheral blood mononuclear cells (PBMCs) may offer insights into liver fibrosis development in MASLD.
Purpose of the Study:
- To explore the association between active DNA demethylation in PBMCs and liver fibrosis in individuals with MASLD.
- To examine DNA methylation and mRNA levels of inflammation- and fibrosis-related genes in relation to fibrosis risk in MASLD.
- To determine if epigenetic alterations in PBMCs can serve as biomarkers for MASLD progression.
Main Methods:
- Quantified global demethylation intermediates (5-hydroxymethylcytosine [5hmC], 5-formylcytosine [5fC]) in PBMCs from 89 participants using ELISA.
- Analyzed site-specific DNA methylation of SOCS3, SREBF1, and TXNIP via mass spectrometry-based bisulfite sequencing.
- Assessed mRNA expression of candidate genes using RT-PCR and correlated findings with the fibrosis non-alcoholic steatohepatitis (NASH) index (FNI).
Main Results:
- Individuals with MASLD and higher fibrosis risk (FNI) showed elevated global 5hmC and 5fC levels in PBMCs.
- Increased FNI correlated with reduced SOCS3 gene methylation.
- Higher FNI was associated with increased mRNA expression of SOCS3, TXNIP, IL-6, and MCP-1.
Conclusions:
- Elevated fibrosis risk in MASLD is linked to active global DNA demethylation in PBMCs.
- Differential methylation and expression of inflammation- and fibrosis-related genes in PBMCs may mirror liver changes.
- These epigenetic alterations in PBMCs could serve as potential biomarkers for MASLD progression and liver fibrogenesis.
