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We found that acetaminophen-induced inflammation exacerbated AAI hepatotoxicity in mice and increased hepatic AAI-related protein adduction. While Mpo-knockout mice exhibited attenuated liver injury and fewer AAI-protein adduction. Further investigation revealed that MPO mediates the metabolic activation of AAI, specifically its metabolite aristolactam I (ALI). An ALI-chloride (ALI-Cl) intermediate was identified in both ALI\/leukocyte incubations and ALI\/hypochlorous acid incubations. ALI-Cl exhibited chemical reactivity toward N-acetylcysteine (NAC) and cysteine, forming ALI-NAC or ALI-cysteine conjugates when ALI was incubated with leukocytes. Additionally, ALI-Cl also reacted with sulfhydryl groups on cysteine residues of cellular protein in leukocytes. After enzymatic hydrolysis, the resultant ALI-cysteine adduct matched that identified in the liver digests of AAI-exposed mice. This study facilitates the understanding of the mechanisms underlying AAI toxicity under inflammatory conditions.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06815","type":"journal-article","created":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T13:26:38Z","timestamp":1784640398000},"page":"23827-23838","source":"Crossref","is-referenced-by-count":0,"title":["Myeloperoxidase-Mediated\nMetabolic Activation of Aristolochic\nAcid I Aggravates Liver Injury Under Inflammatory Conditions"],"prefix":"10.1021","volume":"74","author":[{"given":"Yudi","family":"Jia","sequence":"first","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. China, 110016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanjia","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. 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China, 110016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7921-3727","authenticated-orcid":true,"given":"Weiwei","family":"Li","sequence":"additional","affiliation":[{"name":"Guizhou Medical University , , ,","place":["Guiyang, P. R. China, 550004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zixia","family":"Hu","sequence":"additional","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. China, 110016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2211-7803","authenticated-orcid":true,"given":"Yang","family":"Luan","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University, School of Medicine , , ,","place":["Shanghai, P. R. China, 200025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University, School of Medicine , , ,","place":["Shanghai, P. R. China, 200025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Peng","sequence":"additional","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. China, 110016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Li","sequence":"additional","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. China, 110016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8187-2147","authenticated-orcid":true,"given":"Jiang","family":"Zheng","sequence":"additional","affiliation":[{"name":"Shenyang Pharmaceutical University , , , ,","place":["Shenyang, Liaoning, P. R. China, 110016"]},{"name":"Guizhou Medical University , , ,","place":["Guiyang, P. R. China, 550004"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,21]]},"reference":[{"key":"2026080510090701500_cit1","doi-asserted-by":"publisher","first-page":"648","DOI":"10.3389\/fphar.2019.00648","article-title":"Systematic\nOverview of Aristolochic Acids: Nephrotoxicity, Carcinogenicity, and\nUnderlying Mechanisms","volume":"10","author":"Han","year":"2019","journal-title":"Front. Pharmacol."},{"issue":"1","key":"2026080510090701500_cit2","first-page":"3","article-title":"Ancient medicinal\nuse of Aristolochia: birthwort\u2019s tradition and toxicity","volume":"53","author":"Scarborough","year":"2011","journal-title":"Pharm. Hist."},{"issue":"7150","key":"2026080510090701500_cit3","doi-asserted-by":"publisher","first-page":"148","DOI":"10.1038\/448148a","article-title":"Toxicology:\ndanger in the diet","volume":"448","author":"Lincoln","year":"2007","journal-title":"Nature"},{"issue":"10","key":"2026080510090701500_cit4","doi-asserted-by":"publisher","first-page":"576","DOI":"10.1038\/s41568-022-00494-x","article-title":"Aristolochic acid-associated cancers:\na public health risk in need of global action","volume":"22","author":"Das","year":"2022","journal-title":"Nat. Rev. Cancer"},{"issue":"13","key":"2026080510090701500_cit5","doi-asserted-by":"publisher","first-page":"7438","DOI":"10.1021\/acs.jafc.4c00751","article-title":"Comprehensive HRMS Screening and\nRisk Assessments of Aristolochic Acid Analogues in Asari Radix et\nRhizoma and Related Commercial Health Products","volume":"72","author":"Hu","year":"2024","journal-title":"J. Agric. Food Chem."},{"issue":"5","key":"2026080510090701500_cit6","doi-asserted-by":"publisher","first-page":"522","DOI":"10.1021\/acsagscitech.1c00144","article-title":"Determination\nof Aristolochic\nAcids in Soil, Water, and Herbal Plants in Medicinal Plant Cultivation\nAreas: An Emerging Environmental Contaminant Worth Concerning","volume":"1","author":"Chan","year":"2021","journal-title":"ACS Agric. Sci. Technol."},{"key":"2026080510090701500_cit7","volume-title":"Pharmacopoeia of the People\u2019s Republic\nof China (Vol. I)","author":"Chinese Pharmacopoeia Commission.","year":"2025"},{"key":"2026080510090701500_cit8","doi-asserted-by":"publisher","first-page":"134111","DOI":"10.1016\/j.chemosphere.2022.134111","article-title":"Aristolochic acid\nI as an emerging biogenic contaminant\ninvolved in chronic kidney diseases: A comprehensive review on exposure\npathways, environmental health issues and future challenges","volume":"297","author":"Lukinich-Gruia","year":"2022","journal-title":"Chemosphere"},{"issue":"1","key":"2026080510090701500_cit9","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1021\/acs.jafc.5b05089","article-title":"Uptake and\nAccumulation of Nephrotoxic and Carcinogenic\nAristolochic Acids in Food Crops Grown in Aristolochia clematitis-Contaminated\nSoil and Water","volume":"64","author":"Li","year":"2016","journal-title":"J. Agric. 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Res."},{"issue":"2","key":"2026080510090701500_cit16","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1093\/carcin\/11.2.313","article-title":"Aristolochic acid binds covalently\nto the exocyclic amino group of purine nucleotides in DNA","volume":"11","author":"Pfau","year":"1990","journal-title":"Carcinogenesis"},{"issue":"1","key":"2026080510090701500_cit17","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1021\/acs.chemrestox.0c00454","article-title":"Quantitation of Protein\nAdducts of Aristolochic Acid I by Liquid Chromatography-Tandem Mass\nSpectrometry: A Novel Method for Biomonitoring Aristolochic Acid Exposure","volume":"34","author":"Chan","year":"2021","journal-title":"Chem. Res. Toxicol."},{"issue":"7","key":"2026080510090701500_cit18","doi-asserted-by":"publisher","first-page":"1429","DOI":"10.1038\/s41401-023-01052-3","article-title":"Skullcapflavone\nII, a novel NQO1 inhibitor, alleviates aristolochic acid I-induced\nliver and kidney injury in mice","volume":"44","author":"Dong","year":"2023","journal-title":"Acta Pharmacol.\nSin."},{"issue":"7","key":"2026080510090701500_cit19","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1093\/carcin\/bgw045","article-title":"Sulfotransferase-1A1-dependent\nbioactivation of aristolochic\nacid I and N-hydroxyaristolactam I in human cells","volume":"37","author":"Hashimoto","year":"2016","journal-title":"Carcinogenesis"},{"key":"2026080510090701500_cit20","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.toxlet.2023.02.009","article-title":"Aristolochic\nacid induces an inflammatory response with prostaglandin E2 production\nand apoptosis in NRK-52E proximal tubular cells","volume":"378","author":"Komatsu","year":"2023","journal-title":"Toxicol. Lett."},{"issue":"7","key":"2026080510090701500_cit21","doi-asserted-by":"publisher","first-page":"1578","DOI":"10.1002\/ijc.31544","article-title":"Herbal\nmedicine containing aristolochic acid and the risk of hepatocellular\ncarcinoma in patients with hepatitis B virus infection","volume":"143","author":"Chen","year":"2018","journal-title":"Int. J. Cancer"},{"issue":"1","key":"2026080510090701500_cit22","doi-asserted-by":"publisher","first-page":"132","DOI":"10.3390\/antiox13010132","article-title":"The Roles of Neutrophil-Derived\nMyeloperoxidase (MPO) in Diseases: The New Progress","volume":"13","author":"Lin","year":"2024","journal-title":"Antioxidants (Basel)"},{"issue":"12","key":"2026080510090701500_cit23","doi-asserted-by":"publisher","first-page":"2459","DOI":"10.1021\/acs.chemrestox.9b00280","article-title":"Metabolic Activation of Tofacitinib\nMediated by Myeloperoxidase in Vitro","volume":"32","author":"Guo","year":"2019","journal-title":"Chem.\nRes. Toxicol."},{"issue":"7","key":"2026080510090701500_cit24","doi-asserted-by":"publisher","first-page":"1448","DOI":"10.1161\/ATVBAHA.119.312725","article-title":"Inhibition of MPO (Myeloperoxidase) Attenuates Endothelial\nDysfunction in Mouse Models of Vascular Inflammation and Atherosclerosis","volume":"39","author":"Cheng","year":"2019","journal-title":"Arterioscler Thromb Vasc Biol."},{"issue":"7","key":"2026080510090701500_cit25","doi-asserted-by":"publisher","first-page":"3433","DOI":"10.1021\/acs.jafc.2c07939","article-title":"Inflammation Intensifies Monocrotaline-Induced Liver\nInjury","volume":"71","author":"Gong","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"2026080510090701500_cit26","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1007\/978-1-4615-3796-0_10","article-title":"Myeloperoxidase:\nLocalization, Structure, and Function","volume-title":"Blood Cell Biochemistry Vol. 3: Lymphocytes and Granulocytes","author":"Tobler","year":"1991"},{"issue":"3","key":"2026080510090701500_cit27","doi-asserted-by":"publisher","first-page":"1476","DOI":"10.1016\/S0022-3565(25)12207-6","article-title":"Clozapine is oxidized\nby activated human neutrophils\nto a reactive nitrenium ion that irreversibly binds to the cells","volume":"275","author":"Liu","year":"1995","journal-title":"J. Pharmacol. Exp. Ther."},{"issue":"6","key":"2026080510090701500_cit28","doi-asserted-by":"publisher","first-page":"999","DOI":"10.1016\/S0026-895X(24)13269-5","article-title":"A comparison\nof the covalent binding of clozapine and olanzapine to human neutrophils\nin vitro and in vivo","volume":"53","author":"Gardner","year":"1998","journal-title":"Mol. Pharmacol."},{"issue":"6","key":"2026080510090701500_cit29","doi-asserted-by":"publisher","first-page":"991","DOI":"10.1016\/S0026-895X(24)13268-3","article-title":"A comparison of\nthe oxidation of clozapine and olanzapine to reactive metabolites\nand the toxicity of these metabolites to human leukocytes","volume":"53","author":"Gardner","year":"1998","journal-title":"Mol. Pharmacol."},{"issue":"3","key":"2026080510090701500_cit30","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1016\/S0022-3565(25)22554-X","article-title":"Metabolism\nof vesnarinone by activated\nneutrophils: implications for vesnarinone-induced agranulocytosis","volume":"270","author":"Uetrecht","year":"1994","journal-title":"J. Pharmacol. Exp. Ther."},{"issue":"2","key":"2026080510090701500_cit31","doi-asserted-by":"publisher","first-page":"226","DOI":"10.1021\/tx00044a007","article-title":"Oxidation\nof aminopyrine by hypochlorite to a reactive dication: possible implications\nfor aminopyrine-induced agranulocytosis","volume":"8","author":"Uetrecht","year":"1995","journal-title":"Chem.\nRes. Toxicol."},{"key":"2026080510090701500_cit32","doi-asserted-by":"publisher","first-page":"114642","DOI":"10.1016\/j.jep.2021.114642","article-title":"The genus Asarum: A\nreview on phytochemistry, ethnopharmacology,\ntoxicology and pharmacokinetics","volume":"282","author":"Liu","year":"2022","journal-title":"J. Ethnopharmacol."},{"issue":"6","key":"2026080510090701500_cit33","doi-asserted-by":"publisher","first-page":"971","DOI":"10.1093\/oxfordjournals.jbchem.a022713","article-title":"Role of myeloperoxidase in the neutrophil-induced\noxidation of low\ndensity lipoprotein as studied by myeloperoxidase-knockout mouse","volume":"127","author":"Noguchi","year":"2000","journal-title":"J. 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Neuroinflammation"},{"issue":"8090","key":"2026080510090701500_cit40","doi-asserted-by":"publisher","first-page":"747","DOI":"10.1038\/s41586-025-09523-9","article-title":"Myeloperoxidase\ntransforms chromatin into neutrophil extracellular traps","volume":"647","author":"Burn","year":"2025","journal-title":"Nat."},{"issue":"2","key":"2026080510090701500_cit41","doi-asserted-by":"publisher","first-page":"1243","DOI":"10.3390\/ijms24021243","article-title":"The Role of\nMyeloperoxidase in Clozapine-Induced Inflammation:\nA Mechanistic Update for Idiosyncratic Drug-Induced Agranulocytosis","volume":"24","author":"Sernoskie","year":"2023","journal-title":"Int. J. Mol. Sci."},{"issue":"Pt 2","key":"2026080510090701500_cit42","doi-asserted-by":"publisher","first-page":"122113","DOI":"10.1016\/j.saa.2022.122113","article-title":"Comparison\nof the degradation mechanisms of diclofenac in the presence of iron\noctacarboxyphthalocyanine and myeloperoxidase","volume":"287","author":"Nackiewicz","year":"2023","journal-title":"Spectrochim Acta A Mol. Biomol Spectrosc"},{"issue":"32","key":"2026080510090701500_cit43","doi-asserted-by":"publisher","first-page":"18155","DOI":"10.1021\/acs.jafc.4c03508","article-title":"Positive Feedback Mechanism in Aristolochic\nAcid I Exposure-Induced\nAnemia and DNA Adduct Formation: Implications for Balkan Endemic Nephropathy","volume":"72","author":"Ham","year":"2024","journal-title":"J. Agric. Food Chem."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c06815","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/30\/23827\/66084715\/acs.jafc.6c06815.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/30\/23827\/66084715\/acs.jafc.6c06815.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T14:12:46Z","timestamp":1785939166000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/30\/23827\/5224536\/Myeloperoxidase-Mediated-Metabolic-Activation-of"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,21]]},"references-count":43,"journal-issue":{"issue":"30","published-online":{"date-parts":[[2026,7,21]]},"published-print":{"date-parts":[[2026,8,5]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c06815","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,21]]}}}