基于RNA-seq探讨维生素D3改善代谢相关脂肪性肝病大鼠肝脏损伤的可能机制OA

Objective The aim of this study was to investigate the effects of vitamin D3 on hepatic pathological alterations and gene expression profiles in rats with metabolic dysfunction-associated steatotic liver disease(MASLD).Methods Rats were randomly divided into the control group(CON group,n=12)and the high-fat diet group(HFD group,n=18).After being fed a standard chow or a high-fat diet for 7 weeks,six rats from each group were euthanized to confirm that the model was successfully established.Subse-quently,the remaining HFD-fed rats were randomly assigned to the HFD group(n=6)or the HFD+VD group(n=6),with the HFD+VD group receiving vitamin D3 intervention.At the end of the intervention,all rats were euthanized,and liver tissues were col-lected for analysis.Hematoxylin and eosin(HE)staining,Oil Red O staining,and Masson's trichrome staining were used to evaluate pathological changes in the liver,lipid accumulation,and the degree of fibrosis,respectively.Transcriptome sequencing was employed to analyze the gene expression profiles in liver tissues,and differentially expressed genes(DEGs)were identified.Bioinformatics analy-ses were conducted using KEGG enrichment analysis,gene set enrichment analysis(GSEA),and the construction of protein-protein interaction(PPI)networks.The expression changes of the macrophage markers Cd86 and Cd163 in the liver were detected by immuno-histochemistry(IHC).Intergroup comparisons were performed using analysis of variance(ANOVA).Results The hepatic pathological damage,lipid droplet area,and relative collagen area in the MASLD rats of the HFD+VD group were significantly lower than those in the HFD group(P<0.05).The transcriptome analysis results showed that the differentially expressed genes in the HFD+VD group were significantly enriched in inflammation-related signaling pathways(such as the Toll-like receptor signaling pathway).The PPI net-work analysis revealed that the hub genes identified in the HFD+VD group were associated with immune regulation and signal trans-duction.The immunohistochemistry results indicated that the protein expression level of the M1 macrophage marker Cd86 in the HFD group was significantly higher than that in the CON group(P<0.05),while it was significantly lower in the HFD+VD group than in the HFD group(P<0.05).There was no significant difference in the protein expression levels of Cd163 among the three groups(P>0.05).Conclusion Vitamin D3 may alleviate hepatic injury in MASLD rats by modulating macrophage polarization through modulates inflammatory-related signaling pathways such as the Toll-like receptor signaling pathway.