基于RNA测序技术探究中性粒细胞在酒精性肝病中的作用及机制OA

Aim To explore the role of hepatic neutro-phils in the pathogenesis and progression of alcoholic liver disease(ALD)using RNA sequencing(RNA-seq),and to identify the potential therapeutic targets for ALD prevention and treatment.Methods An alcohol-induced liver injury model was established in mice using the Gao-binge method.Hepatic neutrophils were isolated via in situ perfusion and analyzed by RNA-seq.Bioinformatic analyses,including differen-tial gene expression,gene ontology(GO),Kyoto ency-clopedia of genes and genomes(KEGG),and protein-protein interaction(PPI)network analyses,were per-formed.Quantitative real-time PCR(qRT-PCR)and Western blot were employed to validate key genes.Im-munohistochemistry and immunofluorescence were used to assess neutrophil infiltration and gene expres-sion in liver tissue.Results Compared with controls,neutrophils from ALD mice exhibited significant tran-scriptomic alterations,with 662 differentially ex-pressed genes(419 upregulated and 243 downregu-lated).These genes were mainly enriched in meta-bolic and regulatory pathways.PPI analysis identified S100A4 as a central hub gene,whose mRNA and pro-tein levels markedly increased in ALD neutrophils.Conclusions RNA-seq and integrative bioinformat-ics analyses reveal profound transcriptomic reprogram-ming of hepatic neutrophils in ALD.S100A4 is signifi-cantly upregulated and may contribute to ALD patho-genesis,providing a potential molecular target for its prevention and therapy.