Czech J. Anim. Sci., 2026, 71(7):330-341 | DOI: 10.17221/102/2025-CJAS

The impact of folic acid on genome-wide DNA methylation profiles in the liver of broilers

Yuting Pang, Yujie Zhang, Yunheng Liu, Xiuling Li, Jinyi Xing ORCID...
1 School of Life Sciences, Linyi University, Linyi, P.R. China

Folic acid (FA) plays an important role as a cofactor and coenzyme in animal growth and development, and in the regulation of gene expression and methylation. However, the epigenetic mechanism by which dietary FA supplementation regulates lipid metabolism in broilers remains largely unknown. Therefore, this study aimed to investigate the effects of FA on genome-wide DNA methylation profiles in broiler liver and to identify differentially methylated genes (DMGs) and pathways associated with lipid metabolism. Reduced representation bisulphite sequencing (RRBS) was performed to analyse genome-wide DNA methylation profiles in liver tissues of broilers supplemented with 0 (control, C group), 5 (M group) or 10 mg/kg (H group) FA in basal diet for 42 days. The genome-wide methylation analysis showed that the average percentage of each sample of CpG (5'-C-phosphate-G-3') methylation efficiency was 29.47%. A total of 1 402 differentially methylated regions (DMRs) were detected between C and M groups, including 745 hyper-DMRs and 657 hypo-DMRs, Compared with the C group, 410 hyper-DMRs and 436 hypo-DMRs were identified in the H group. Additionally, 511 hyper-DMRs and 560 hypo-DMRs were found between the M and H group. Furthermore, a total of 134 DMGs exhibited DMRs in the promoter region across all three comparison groups, including 27 upregulated DMGs and 20 downregulated DMGs. Gene Ontology (GO) analysis of DMRs revealed that the associated genes are mainly involved in H3-K4 demethylation, histone lysine demethylation, peptidyl-lysine methylation, histone demethylation, lysine N-methyltransferase activity, fat cell proliferation, cellular lipid metabolic processes, fatty-acyl-CoA binding. In addition, key Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathways were primarily associated with the Toll-like receptor signalling pathway, fatty acid metabolism, butanoate metabolism, retinol metabolism. Taken together, these results demonstrated that FA supplementation affects genome-wide DNA methylation profiles in the liver of broilers, suggesting an underlying epigenetic mechanism for its regulation of hepatic lipid metabolism.

Keywords: broilers; DNA methylation; folic acid; lipid metabolism; liver

Received: July 14, 2025; Accepted: June 17, 2026; Prepublished online: July 24, 2026; Published: July 28, 2026  Show citation

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Pang Y, Zhang Y, Liu Y, Li X, Xing J. The impact of folic acid on genome-wide DNA methylation profiles in the liver of broilers. Czech Journal of Animal Science. 2026;71(7):330-341. doi: 10.17221/102/2025-CJAS.
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