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Although the clinical efficacy of berberine (BBR) in treating NAFLD has been established, the mechanism remains uncertain. This study is to evaluate the effects of BBR on hepatic LDs and investigate the underlying mechanisms. Using high\u2010fat diet\u2010induced obese (DIO) mice as the model for NAFLD, BBR was administered daily by gavage for 4\u2009weeks. Liver tissue was examined for changes in lipid deposition and histology. Transcriptomics was performed to screen differently expressed genes. The potential targets of BBR against NAFLD were then determined by Western Blot and immunostaining. In oleic acid (OA)\u2010induced HepG2 cells, the link between BBR and potential targets was further elucidated through the activation or antagonism of PPAR\u03b1. The binding of BBR to potential targets was predicted using molecular docking. BBR significantly reduced hepatic steatosis by decreasing LD size rather than number. Transcriptomics with validation demonstrated that BBR modulated the expression of LD\u2010associated proteins CIDEA and PLIN4 in the liver. Further investigations revealed that BBR reversed the abnormal elevation of BSCL2 and PLIN2 in steatotic livers. Finally, we found that BBR reduced LD size in OA\u2010induced HepG2 cells by regulating BSCL2 and PPAR\u03b1\u2010mediated CIDEA\/PLIN4\/PLIN2. Notably, BBR could bind well to PPAR\u03b1 and BSCL2. BBR can attenuate hepatic steatosis in DIO mice by reducing LD size through the regulation of LD\u2010associated proteins.<\/jats:p>","DOI":"10.1111\/jcmm.70524","type":"journal-article","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T23:48:24Z","timestamp":1744069704000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Berberine Attenuates Nonalcoholic Hepatic Steatosis by Regulating Lipid Droplet\u2010Associated Proteins: In\u00a0Vivo, In\u00a0Vitro and Molecular Evidence"],"prefix":"10.1111","volume":"29","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-0248-4465","authenticated-orcid":false,"given":"Hongzhan","family":"Wang","sequence":"first","affiliation":[{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Tongji Hospital Tongji Medical College, Huazhong University of Science and Technology  Wuhan 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