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J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Acetaminophen (APAP) represents a major cause of drug-induced liver injury (DILI), and effective pharmacological interventions remain limited. Annexin A5 (AnxA5), a Ca2+-dependent phospholipid-binding protein, participates in diverse biological processes related to tissue repair. In this study, we evaluated AnxA5 expression in APAP-challenged mouse livers and clinical samples from patients with liver injury. Using hepatic cell lines AML12 and HepG2, we performed overexpression-based functional assays to assess the cytoprotective effects of AnxA5 against APAP toxicity. Co-immunoprecipitation assays were applied to characterize protein interactions, and mitochondrial functional parameters were measured to dissect the underlying molecular mechanism. We found that AnxA5 was significantly upregulated in both APAP exposed mice and APAP DILI patients. Cellular functional assays showed that AnxA5 overexpression mitigated APAP triggered cytotoxicity in AML12 and HepG2 cells. Mechanistically, AnxA5 bound to voltage dependent anion channel 1 (VDAC1), restrained VDAC1 mediated mitochondrial Ca2+ influx, and suppressed VDAC1 oligomerization, which further inhibited mitochondrial permeability transition pore (mPTP) opening. In an APAP-induced liver injury mouse model, exogenous recombinant AnxA5 treatment maintained mitochondrial integrity and ameliorated hepatic inflammation and liver damage. Collectively, our data reveal AnxA5 as an endogenous mitochondrial protective factor and support its therapeutic potential against APAP-induced liver injury.<\/jats:p>","DOI":"10.3390\/ijms27177771","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:48:36Z","timestamp":1788256116000},"page":"7771","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Annexin A5 Maintains Mitochondrial Integrity by Inhibiting mPTP Opening to Protect Against Acetaminophen-Induced Acute Liver Injury"],"prefix":"10.3390","volume":"27","author":[{"given":"Xiaowen","family":"Zhang","sequence":"first","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Wenwei","family":"Li","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Luqi","family":"Li","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"}]},{"given":"Wei","family":"Tang","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"software","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3733-8104","authenticated-orcid":false,"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"conceptualization","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Zichun","family":"Hua","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China"},{"name":"Changzhou High-Tech Research Institute of Nanjing University, Changzhou 213164, China"},{"name":"Faculty of Pharmaceutical Sciences, Xinxiang Medical University, Xinxiang 453002, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"R1147","DOI":"10.1016\/j.cub.2017.09.019","article-title":"The liver","volume":"27","author":"Trefts","year":"2017","journal-title":"Curr. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1016\/j.bbadis.2018.08.037","article-title":"Animal models of drug-induced liver injury","volume":"1865","author":"McGill","year":"2019","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_3","first-page":"131","article-title":"Acetaminophen-Induced Hepatotoxicity: A Comprehensive Update","volume":"4","author":"Yoon","year":"2016","journal-title":"J. Clin. Transl. Hepatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3377","DOI":"10.1007\/s00204-021-03142-9","article-title":"Delayed administration of N-acetylcysteine blunts recovery after an acetaminophen overdose unlike 4-methylpyrazole","volume":"95","author":"Akakpo","year":"2021","journal-title":"Arch. Toxicol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"760","DOI":"10.1111\/liv.15822","article-title":"The integrated stress response-related expression of CHOP due to mitochondrial toxicity is a warning sign for DILI liability","volume":"44","author":"Vlasveld","year":"2024","journal-title":"Liver Int."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1093\/toxsci\/kfae055","article-title":"Drug-induced impairment of mitochondrial fatty acid oxidation and steatosis: Assessment of causal relationship with 45 pharmaceuticals","volume":"200","author":"Buron","year":"2024","journal-title":"Toxicol. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"101115","DOI":"10.1016\/j.gendis.2023.101115","article-title":"Mitochondrial dysfunction: A promising therapeutic target for liver diseases","volume":"11","author":"Chen","year":"2024","journal-title":"Genes. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1146\/annurev-pathmechdis-051122-094016","article-title":"Acetaminophen Hepatotoxicity: Paradigm for Understanding Mechanisms of Drug-Induced Liver Injury","volume":"19","author":"Jaeschke","year":"2024","journal-title":"Annu. Rev. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2034","DOI":"10.1074\/mcp.RA118.000875","article-title":"Reactive metabolite-induced protein glutathionylation: A potentially novel mechanism underlying acetaminophen hepatotoxicity","volume":"17","author":"Chan","year":"2018","journal-title":"Mol. Cell. Proteom."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5534","DOI":"10.1021\/bi992779z","article-title":"Annexin V binding perturbs the cardiolipin fluidity gradient in isolated mitochondria. Can it affect mitochondrial function?","volume":"39","author":"Megli","year":"2000","journal-title":"Biochemistry"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"110591","DOI":"10.1016\/j.biopha.2020.110591","article-title":"Roles of Annexin A protein family in autophagy regulation and therapy","volume":"130","author":"Xi","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_12","first-page":"3413","article-title":"Annexin A5 controls VDAC1-dependent mitochondrial Ca2+ homeostasis and determines cellular susceptibility to apoptosis","volume":"44","author":"Trenker","year":"2025","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8054","DOI":"10.1073\/pnas.132598099","article-title":"DT40 cells lacking the Ca2+-binding protein annexin 5 are resistant to Ca2+-dependent apoptosis","volume":"99","author":"Hawkins","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2469","DOI":"10.1074\/jbc.M113.450163","article-title":"Role of annexin A5 in cisplatin-induced toxicity in renal cells: Molecular mechanism of apoptosis","volume":"289","author":"Jeong","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Frosteg\u00e5rd, A., and Haegerstrand, A. (2024). New Therapeutic Strategies in Retinal Vascular Diseases: A Lipid Target, Phosphatidylserine, and Annexin A5-A Future Theranostic Pairing in Ophthalmology. Pharmaceuticals, 17.","DOI":"10.3390\/ph17080979"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"424","DOI":"10.2119\/molmed.2016.00026","article-title":"Annexin A5 increases survival in murine sepsis model by inhibiting HMGB1-mediated pro-inflammation and coagulation","volume":"22","author":"Park","year":"2016","journal-title":"Mol. Med."},{"key":"ref_17","unstructured":"Zhu, Z., He, J., Liu, Q., Guo, J., Liu, J., Xu, S., Chen, X., and Wu, D. (2025). Annexin A5 in Ischemic Stroke: Molecular Mechanisms and Translational Therapy Strategies. Aging Dis., Online ahead of print."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Zhao, S., Feng, Y., Zhang, J., Zhang, Q., Wang, J., and Cui, S. (2024). Comparative analysis of gene expression between mice and humans in acetaminophen-induced liver injury by integrating bioinformatics analysis. BMC Med. Genom., 17.","DOI":"10.1186\/s12920-024-01848-0"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1002\/1873-3468.14078","article-title":"Calcium transfer between endoplasmic reticulum and mitochondria in liver diseases","volume":"595","author":"Jin","year":"2021","journal-title":"FEBS Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.ceca.2017.06.007","article-title":"VDAC1 functions in Ca2+ homeostasis and cell life and death in health and disease","volume":"69","author":"Krelin","year":"2018","journal-title":"Cell Calcium"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4665530","DOI":"10.1155\/2022\/4665530","article-title":"The Potential Role of Voltage-Dependent Anion Channel in the Treatment of Parkinson\u2019s Disease","volume":"2022","author":"He","year":"2022","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e56297","DOI":"10.15252\/embr.202256297","article-title":"MIRO-1 interacts with VDAC-1 to regulate mitochondrial membrane potential in Caenorhabditis elegans","volume":"24","author":"Ren","year":"2023","journal-title":"EMBO Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"52623","DOI":"10.1074\/jbc.M409009200","article-title":"Cell surface-expressed phosphatidylserine and annexin A5 open a novel portal of cell entry","volume":"279","author":"Kenis","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1038\/s41392-021-00626-z","article-title":"Phosphatidylserine externalized on the colonic capillaries as a novel pharmacological target for IBD therapy","volume":"6","author":"Zhang","year":"2021","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1080\/03602532.2020.1832112","article-title":"The development and hepatotoxicity of acetaminophen: Reviewing over a century of progress","volume":"52","author":"McGill","year":"2020","journal-title":"Drug Metab. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.redox.2018.04.019","article-title":"Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions","volume":"17","author":"Yan","year":"2018","journal-title":"Redox Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"143","DOI":"10.14715\/cmb\/2024.70.7.20","article-title":"99mTc-HYNIC-Annexin V detection of apoptosis in the rabbit model of liver cancer","volume":"70","author":"Shi","year":"2024","journal-title":"Cell Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Cheng, Q., Lu, L., Grafstr\u00f6m, J., Olofsson, M.H., Thorell, J.-O., Sam\u00e9n, E., Johansson, K., Ahlz\u00e9n, H.-S., Stone-Elander, S., and Linder, S. (2012). Combining [11C]-AnxA5 PET imaging with serum biomarkers for improved detection in live mice of modest cell death in human solid tumor xenografts. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0042151"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2859","DOI":"10.1007\/s00018-007-7297-2","article-title":"Annexin A5: Shifting from a diagnostic towards a therapeutic realm","volume":"64","author":"Kenis","year":"2007","journal-title":"Cell Mol. Life Sci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Grewal, T., Rentero, C., Enrich, C., Wahba, M., Raabe, C.A., and Rescher, U. (2021). Annexin Animal Models-From Fundamental Principles to Translational Research. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073439"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"215857","DOI":"10.1016\/j.canlet.2022.215857","article-title":"Annexin A5 as a targeting agent for cancer treatment","volume":"547","author":"Woodward","year":"2022","journal-title":"Cancer Lett."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7771\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T04:41:18Z","timestamp":1788496878000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7771"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,30]]},"references-count":31,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27177771"],"URL":"https:\/\/doi.org\/10.3390\/ijms27177771","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,30]]}}}