基于药物重定位发现抗代谢相关脂肪性肝炎肝纤维化先导物甲苯咪唑OA

Aim To screen and validate lead com-pounds with anti-fibrotic effects on metabolic dysfunction-associated steatohepatitis(MASH)and to provide scientific data for the development of novel MASH therapeutic drugs based on a drug repurposing strategy.Methods We used R programming and the online bioinformatics platform Galaxy to analyze the differentially expressed genes(DEGs)between the treatment and model groups in human liver organoids and mouse liver tissues treated with the approved drug resmetirom.Potential therapeutic compounds for MASH-associated liver fibrosis were predicted using the Connectivity Map(CMap)database.Cell viability was assessed by MTT assay.Lipid accumulation in he-patocytes was evaluated by Oil Red O staining.The ex-pression of fibrosis-related proteins in hepatic stellate cells was examined by Western blotting,while lipid metabolism-related genes were quantified by qRT-PCR.The binding affinity between candidate com-pounds and thyroid hormone receptor(THR)was as-sessed using molecular docking,and THR activation was evaluated using a dual-luciferase reporter assay.Results The bioinformatics analysis showed that 366 and 627 DEGs were identified in the two datasets,re-spectively.GO and KEGG enrichment analysis re-vealed that the DEGs were associated with MASH-related liver fibrosis.CMap analysis identified five FDA-approved drugs,including mebendazole,as po-tential anti-liver fibrosis agents.Mebendazole exhib-ited favorable safety profiles in hepatocytes,signifi-cantly reduced intracellular lipid accumulation,and downregulated fibrosis-related factors induced by TGF-β and PA in LX-2 cells.Mebendazole activated THR-β,thereby upregulating the expression of FA β-oxida-tion and lipolysis related genes.Conclusion Meben-dazole could activate THR-β,modulate cellular lipid metabolism,inhibit hepatic steatosis,and suppress TGF-β and PA-induced activation of hepatic stellate cells.