{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T22:29:11Z","timestamp":1791930551758,"version":"4.3.4"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T00:00:00Z","timestamp":1756771200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hunan Provincial Department of Education Research Project","award":["23B0375"],"award-info":[{"award-number":["23B0375"]}]},{"name":"Hunan Provincial Administration of Traditional Chinese Medicine Research Project","award":["C2023021"],"award-info":[{"award-number":["C2023021"]}]},{"name":"Hunan University of Traditional Chinese Medicine University-level Research Project","award":["2024XJZC017"],"award-info":[{"award-number":["2024XJZC017"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Huanglian Wendan Decoction (HLWDD), a classical traditional Chinese medicine (TCM) formula, has shown therapeutic promise in treating metabolic disorders. However, its underlying mechanisms against non-alcoholic fatty liver disease (NAFLD) remain unclear.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>This study aimed to elucidate the pharmacological mechanisms by which HLWDD ameliorates NAFLD, focusing on its impact on lipid metabolism, gut microbiota, and amino acid regulation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A NAFLD rat model was established by administering a high-sugar, high-fat, high-salt diet for 20 weeks. The core components of HLWDD were identified and quantified using UPLC-Q-TOF-MS\/MS and HPLC, and further validated via network pharmacology and molecular docking. Therapeutic efficacy was assessed through analysis of body weight, serum lipid profiles, inflammatory cytokines, hepatic histology, and protein expression. Gut microbiota composition and liver-intestine metabolite profiles were evaluated using metagenomic sequencing and LC-MS\/MS.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Seven key constituents, including quercetin and berberine, were quantified (15.11\u2013164.37 \u03bcg\/mL) and shown to interact with lipid metabolism targets such as liver kinase B1 (LKB1), AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor alpha (PPAR\u03b1), and carnitine palmitoyltransferase 1A (CPT1A). HLWDD treatment significantly reduced body weight, hepatic lipid accumulation, and serum levels of triglycerides, total cholesterol, and low-density lipoprotein cholesterol, while increasing high-density lipoprotein cholesterol. Proinflammatory cytokines (IL-6, IL-1\u03b2, TNF-\u03b1) were notably suppressed. Mechanistically, HLWDD activated the LKB1\/AMPK signaling pathway and modulated aspartic acid metabolism in association with increased abundance of\n                      <jats:italic>\n                        <jats:italic>Akkermansia<\/jats:italic>\n                      <\/jats:italic>\n                      in the gut. Metabolomic analysis identified 13 differential metabolites, with aspartic acid showing strong correlations with\n                      <jats:italic>\n                        <jats:italic>Akkermansia<\/jats:italic>\n                      <\/jats:italic>\n                      and LKB1\/AMPK activity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec005\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      HLWDD exerts its anti-NAFLD effects by enhancing\n                      <jats:italic>\n                        <jats:italic>Akkermansia<\/jats:italic>\n                      <\/jats:italic>\n                      -mediated aspartate metabolism, thereby activating the LKB1\/AMPK axis and promoting lipid oxidation via CPT1A and PPAR\u03b1. This study provides new mechanistic insight into the gut\u2013liver axis in NAFLD and highlights HLWDD as a multi-targeted therapeutic approach for restoring metabolic balance.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0331303","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T17:31:38Z","timestamp":1756834298000},"page":"e0331303","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["Mechanism of Huanglian Wendan Decoction in ameliorating non-alcoholic fatty liver disease via modulating gut microbiota-mediated metabolic reprogramming and activating the LKB1\/AMPK 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Hepatol"},{"issue":"6","key":"pone.0331303.ref006","doi-asserted-by":"crossref","DOI":"10.1016\/j.cgh.2021.11.026","article-title":"Systematic review and meta-analysis: the role of diet in the development of nonalcoholic fatty liver disease","volume":"21","author":"E Tsompanaki","year":"2023","journal-title":"Clin Gastroenterol Hepatol"},{"issue":"3","key":"pone.0331303.ref007","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1152\/physrev.00026.2008","article-title":"LKB1 and AMPK family signaling: the intimate link between cell polarity and energy metabolism","volume":"89","author":"M Jansen","year":"2009","journal-title":"Physiol Rev"},{"issue":"12","key":"pone.0331303.ref008","doi-asserted-by":"crossref","first-page":"1617","DOI":"10.1038\/cr.2012.108","article-title":"LKB1-AMPK axis revisited","volume":"22","author":"F Kottakis","year":"2012","journal-title":"Cell 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Reagan-Shaw","year":"2008","journal-title":"FASEB J"},{"issue":"5","key":"pone.0331303.ref021","article-title":"Traditional Chinese medicine for premature ventricular contraction caused by obstructive sleep apnea: a case report and literature review","volume":"104","author":"Q-Q Yue","year":"2025","journal-title":"Medicine (Baltimore)"},{"issue":"1","key":"pone.0331303.ref022","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1007\/s40520-024-02753-3","article-title":"The impact of the Mediterranean diet on immune function in older adults","volume":"36","author":"F Ecarnot","year":"2024","journal-title":"Aging Clin Exp Res"},{"issue":"12","key":"pone.0331303.ref023","doi-asserted-by":"crossref","first-page":"1596","DOI":"10.1038\/s42255-021-00501-9","article-title":"Non-alcoholic fatty liver disease: the interplay between metabolism, microbes and immunity","volume":"3","author":"H Tilg","year":"2021","journal-title":"Nat Metab"},{"key":"pone.0331303.ref024","doi-asserted-by":"crossref","first-page":"119890","DOI":"10.1016\/j.jep.2025.119890","article-title":"p-Synephrine ameliorates non-alcoholic fatty liver disease by regulating liver-adipose axis via AMPK\/NF-kappa B pathway","volume":"348","author":"W-F Cai","year":"2025","journal-title":"J Ethnopharmacol"},{"issue":"11","key":"pone.0331303.ref025","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1016\/j.cmet.2022.09.017","article-title":"Gut-liver axis: Pathophysiological concepts and clinical implications","volume":"34","author":"H Tilg","year":"2022","journal-title":"Cell Metab"},{"issue":"12","key":"pone.0331303.ref026","doi-asserted-by":"crossref","first-page":"5769","DOI":"10.1002\/ptr.8020","article-title":"Natural compounds against nonalcoholic fatty liver disease: a review on the involvement of the LKB1\/AMPK signaling pathway","volume":"37","author":"N Omidkhoda","year":"2023","journal-title":"Phytother Res"},{"issue":"2","key":"pone.0331303.ref027","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1007\/s10555-023-10156-5","article-title":"Regulation and targeting of SREBP-1 in hepatocellular carcinoma","volume":"43","author":"F Su","year":"2024","journal-title":"Cancer Metastasis Rev"},{"issue":"18","key":"pone.0331303.ref028","doi-asserted-by":"crossref","first-page":"10391","DOI":"10.1021\/acs.jafc.4c00590","article-title":"Molecular mechanisms of fucoxanthin in alleviating lipid deposition in metabolic associated fatty liver disease","volume":"72","author":"S Yang","year":"2024","journal-title":"J Agric Food Chem"},{"issue":"5","key":"pone.0331303.ref029","doi-asserted-by":"crossref","first-page":"1020","DOI":"10.1136\/gutjnl-2021-326789","article-title":"Gut microbiome and health: mechanistic insights","volume":"71","author":"WM de Vos","year":"2022","journal-title":"Gut"},{"issue":"10","key":"pone.0331303.ref030","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1016\/j.clnu.2022.08.029","article-title":"Recent findings in Akkermansia muciniphila-regulated metabolism and its role in intestinal diseases","volume":"41","author":"M-J Liu","year":"2022","journal-title":"Clin Nutr"},{"issue":"1","key":"pone.0331303.ref031","first-page":"1","article-title":"Gut Akkermansia muciniphila ameliorates metabolic dysfunction-associated fatty liver disease by regulating the metabolism of L-aspartate via gut-liver axis","volume":"13","author":"Y Rao","year":"2021","journal-title":"Gut Microbes"},{"key":"pone.0331303.ref032","doi-asserted-by":"crossref","first-page":"1829","DOI":"10.1038\/ncomms2852","article-title":"The gut microbiota suppresses insulin-mediated fat accumulation via the short-chain fatty acid receptor GPR43","volume":"4","author":"I Kimura","year":"2013","journal-title":"Nat Commun"},{"issue":"1","key":"pone.0331303.ref033","doi-asserted-by":"crossref","first-page":"1755","DOI":"10.1038\/s41467-020-15573-6","article-title":"p53-mediated control of aspartate-asparagine homeostasis dictates LKB1 activity and modulates cell survival","volume":"11","author":"L Deng","year":"2020","journal-title":"Nat Commun"},{"issue":"33","key":"pone.0331303.ref034","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2100295118","article-title":"SLC15A4 mediates M1-prone metabolic shifts in macrophages and guards immune cells from metabolic stress","volume":"118","author":"T Kobayashi","year":"2021","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"5","key":"pone.0331303.ref035","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1016\/j.jhep.2017.01.022","article-title":"NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice","volume":"66","author":"AR Mridha","year":"2017","journal-title":"J 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