{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:34Z","timestamp":1787028574257,"version":"build-2736575974"},"reference-count":30,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T00:00:00Z","timestamp":1759708800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["IJMS"],"abstract":"<jats:p>Bisphenol AF (BPAF) exposure is increasingly linked to metabolic disorders, yet the molecular initiating events (MIE) and key events (KE) leading to hepatic steatosis remain unclear. We constructed an adverse outcome pathway (AOP) to mechanistically connect BPAF-triggered macrophage\u2013hepatocyte crosstalk to liver fat accumulation. Male C57BL\/6 mice received daily oral gavage of 0, 0.5, 4, or 32 mg kg\u22121 BPAF for 90 d, and Transwell co-cultures of RAW264.7 macrophages and AML12 hepatocytes were used for in vitro validation. Targeted metabolomics, western blotting, and lipid staining quantified succinate, pathway proteins, and steatosis. BPAF dose-dependently increased serum succinate (BMD = 6901.95 nM) and hepatic triglyceride (TG) (BMD = 874.26 nM). Cryo-EM docking revealed BPAF binding to SUCNR1 at 2.9 \u00c5, disrupting the inactive-state conformation. In co-culture, BPAF-exposed macrophages released succinate that bound hepatocyte SUCNR1, suppressed Akt phosphorylation, and activated JNK. These KEs led to a 40% increase in lipid droplets and elevated TG, total cholesterol (TC), and free fatty acids (FFA) without liver weight gain. We propose the first AOP for BPAF-induced hepatic steatosis: BPAF\u2013SUCNR1 binding (MIE) \u2192 macrophage succinate release (KE1) \u2192 SUCNR1-mediated Akt inhibition\/JNK activation (KE2\u20134) \u2192 hepatic lipid accumulation (KE5) \u2192 steatosis (AO). These findings provide mechanistic insight for chemical risk assessment of BPAF and structurally related bisphenols.<\/jats:p>","DOI":"10.3390\/ijms26199720","type":"journal-article","created":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T15:05:06Z","timestamp":1759763106000},"page":"9720","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Bisphenol AF Induces Hepatic Steatosis via Succinate\u2013SUCNR1-Mediated Macrophage\u2013Hepatocyte Interactions: An Adverse Outcome Pathway Study in Male C57BL\/6 Mice"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5330-2218","authenticated-orcid":false,"given":"Ning","family":"Wang","sequence":"first","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Leng","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huimin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Xu","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoqi","family":"Yu","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelei","family":"Qian","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiqing","family":"Zheng","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengchao","family":"Ying","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gonghua","family":"Tao","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Xiao","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Hong","sequence":"additional","affiliation":[{"name":"Shanghai Municipal Center for Disease Control and Prevention, Institute of Chemical Safety Evaluation\/Key Laboratory of Environmental and Health Impact Assessment of New Pollutants, 1399 at Shenhong Road, Shanghai 201107, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, H., Gao, J., Liu, Y., Ding, Y., Guo, Y., Wang, Z., Dong, Z., and Zhang, N. 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