{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:53:17Z","timestamp":1788432797885,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Afyonkarahisar Health Sciences University Scientific Research Project Unit","award":["24. GENEL.031"],"award-info":[{"award-number":["24. GENEL.031"]}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["IJMS"],"abstract":"<jats:p>Methotrexate (MTX), an effective immunosuppressive and antiproliferative agent, is clinically restricted by its hepatotoxic potential through oxidative stress, inflammation, and apoptosis. Dapagliflozin (DAPA), a sodium\u2013glucose cotransporter 2 inhibitor, exhibits antioxidant and anti-inflammatory actions. This study investigated the hepatoprotective effects of DAPA against MTX-induced acute liver injury. Thirty-two female Wistar albino rats were divided into four groups (n = 8): Control, MTX (20 mg\/kg), MTX + DAPA (MTX + DAPA 10 mg\/kg\/day for 10 days), and DAPA. Liver samples were examined histologically, immunohistochemically (Nuclear factor NF-kappa-B p65 subunit (NF-\u03baB p65), Tumor necrosis factor alpha (TNF-\u03b1), Interleukin 1 beta (IL-1\u03b2), Caspase (Cas)-3, Vascular endothelial growth factor (VEGF)), molecularly (Reverse transcription\u2013polymerase chain for Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), Cytochrome C (Cyt-C), Apoptotic peptidase activating factor 1 (Apaf-1), Cas-9, Cas-3, Cas-12), and biochemically (total oxidant status (TOS), total antioxidant status (TAS) and oxidative stress index (OSI)). MTX induced severe hepatic injury with congestion, sinusoidal dilatation, and inflammatory infiltration, accompanied by upregulation of NF-\u03baB, TNF-\u03b1, IL-1\u03b2, Bax, Cyt-C, Apaf-1, Cas-9, Cas-3, and Cas-12 and reduced Bcl-2. DAPA co-treatment significantly restored hepatic structure, suppressed inflammatory and apoptotic markers, and normalized VEGF expression, indicating reduced pathological angiogenesis. Although DAPA did not fully reverse MTX-induced weight loss, it effectively mitigated hepatocellular damage. DAPA protects against MTX-induced liver injury by inhibiting NF-\u03baB\/TNF-\u03b1\/IL-1\u03b2-mediated inflammation, modulating Bax\/Bcl-2\u2013Cyt-C\u2013Cas-dependent apoptosis, and balancing VEGF-driven angiogenesis. DAPA may thus serve as a promising hepatoprotective adjunct in MTX therapy.<\/jats:p>","DOI":"10.3390\/ijms27052110","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T16:58:18Z","timestamp":1771952298000},"page":"2110","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Cellular Anti-Apoptotic Effects of Dapagliflozin in Methotrexate-Induced Liver Toxicity: Bax\/Bcl-2\/Cyt-C\/Cas-9\/Cas-3 Signaling Pathway"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4671-9315","authenticated-orcid":false,"given":"Emine","family":"Sarman","sequence":"first","affiliation":[{"name":"Department of Histology and Embryology, Faculty of Medicine, Afyonkarahisar Health Sciences University, Afyonkarahisar 03030, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1545-035X","authenticated-orcid":false,"given":"Halil","family":"Asci","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Faculty of Medicine, Suleyman Demirel University, Isparta 32260, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/j.hoc.2012.02.002","article-title":"The Antifolates","volume":"26","author":"Visentin","year":"2012","journal-title":"Hematol. Oncol. Clin. North Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1111\/1751-2980.12184","article-title":"A Review of Methotrexate-associated Hepatotoxicity","volume":"15","author":"Bath","year":"2014","journal-title":"J. Dig. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.1038\/s41375-021-01189-2","article-title":"Folate Metabolism: A Re-Emerging Therapeutic Target in Haematological Cancers","volume":"35","author":"Zarou","year":"2021","journal-title":"Leukemia"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"152840","DOI":"10.1016\/j.tox.2021.152840","article-title":"Hepatotoxic Potentials of Methotrexate: Understanding the Possible Toxicological Molecular Mechanisms","volume":"458","author":"Ezhilarasan","year":"2021","journal-title":"Toxicology"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Hasan Khudhair, D., Al-Gareeb, A.I., Al-kuraishy, H.M., El-Kadem, A.H., Elekhnawy, E., Negm, W.A., Saber, S., Cavalu, S., Tirla, A., and Alotaibi, S.S. (2022). Combination of Vitamin C and Curcumin Safeguards Against Methotrexate-Induced Acute Liver Injury in Mice by Synergistic Antioxidant Effects. Front. Med., 9.","DOI":"10.3389\/fmed.2022.866343"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Schmidt, S., Messner, C.J., Gaiser, C., H\u00e4mmerli, C., and Suter-Dick, L. (2022). Methotrexate-Induced Liver Injury Is Associated with Oxidative Stress, Impaired Mitochondrial Respiration, and Endoplasmic Reticulum Stress In Vitro. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232315116"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"21","DOI":"10.2147\/OTT.S178510","article-title":"Methotrexate-Induced Apoptosis in Human Ovarian Adenocarcinoma SKOV-3 Cells via ROS-Mediated Bax\/Bcl-2-Cyt-c Release Cascading","volume":"12","author":"AlBasher","year":"2018","journal-title":"Onco. Targets. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3479","DOI":"10.1007\/s11033-023-08286-8","article-title":"Morin Ameliorates Methotrexate-Induced Hepatotoxicity via Targeting Nrf2\/HO-1 and Bax\/Bcl2\/Caspase-3 Signaling Pathways","volume":"50","author":"Caglayan","year":"2023","journal-title":"Mol. Biol. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1038\/s41392-023-01570-w","article-title":"Endoplasmic Reticulum Stress: Molecular Mechanism and Therapeutic Targets","volume":"8","author":"Chen","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Pu, S., Pan, Y., Zhang, Q., You, T., Yue, T., Zhang, Y., and Wang, M. (2023). Endoplasmic Reticulum Stress and Mitochondrial Stress in Drug-Induced Liver Injury. Molecules, 28.","DOI":"10.3390\/molecules28073160"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4887","DOI":"10.1007\/s00018-019-03148-8","article-title":"Proinflammatory NFkB Signalling Promotes Mitochondrial Dysfunction in Skeletal Muscle in Response to Cellular Fuel Overloading","volume":"76","author":"Nisr","year":"2019","journal-title":"Cell Mol. Life Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e14299","DOI":"10.7717\/peerj.14299","article-title":"Hypoxia-Induced Factor and Its Role in Liver Fibrosis","volume":"10","author":"Hai","year":"2022","journal-title":"PeerJ"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Darmadi, D., Habriel Ruslie, R., and Pakpahan, C. (2022). Vascular Endothelial Growth Factor (VEGF) in Liver Disease. Tumor Angiogenesis and Modulators, IntechOpen.","DOI":"10.5772\/intechopen.103113"},{"key":"ref_14","first-page":"561","article-title":"Dapagliflozin Prevents Reproductive Damage Caused by Acute Systemic Inflammation through Antioxidant, Anti-inflammatory, and Antiapoptotic Mechanisms","volume":"135","author":"Topsakal","year":"2024","journal-title":"Basic Clin. Pharmacol. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"865","DOI":"10.33594\/000000178","article-title":"Empagliflozin and Dapagliflozin Reduce ROS Generation and Restore NO Bioavailability in Tumor Necrosis Factor \u03b1-Stimulated Human Coronary Arterial Endothelial Cells","volume":"53","author":"Uthman","year":"2019","journal-title":"Cell Physiol. Biochem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1186\/s12933-024-02481-y","article-title":"Dapagliflozin Mitigates Cellular Stress and Inflammation through PI3K\/AKT Pathway Modulation in Cardiomyocytes, Aortic Endothelial Cells, and Stem Cell-Derived \u03b2 Cells","volume":"23","author":"Alsereidi","year":"2024","journal-title":"Cardiovasc. Diabetol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"116991","DOI":"10.1016\/j.taap.2024.116991","article-title":"Dapagliflozin Dampens Liver Fibrosis Induced by Common Bile Duct Ligation in Rats Associated with the Augmentation of the Hepatic Sirt1\/AMPK\/PGC1\u03b1\/FoxO1 Axis","volume":"489","author":"Hassan","year":"2024","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"101821","DOI":"10.1016\/j.toxrep.2024.101821","article-title":"Accidental Methotrexate Overdose Leading to Multisystem Toxicity: A Case Report","volume":"13","author":"Asaduzzaman","year":"2024","journal-title":"Toxicol. Reports"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"9990692","DOI":"10.1155\/jt\/9990692","article-title":"Mitigation of Methotrexate-Induced Intestinal Mucositis in Male Wistar Rats by Gallic Acid: The Role of HGF and C-Met Genes","volume":"2025","author":"Norouzirad","year":"2025","journal-title":"J. Toxicol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1038\/s41467-024-46114-0","article-title":"Inhibition of Mitochondrial Folate Metabolism Drives Differentiation through MTORC1 Mediated Purine Sensing","volume":"15","author":"Zarou","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"103455","DOI":"10.1016\/j.redox.2024.103455","article-title":"Macrophage Dvl2 Deficiency Promotes NOD1-Driven Pyroptosis and Exacerbates Inflammatory Liver Injury","volume":"79","author":"Qu","year":"2025","journal-title":"Redox Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.cotox.2017.11.002","article-title":"NF-\u039aB in Oxidative Stress","volume":"7","author":"Lingappan","year":"2018","journal-title":"Curr. Opin. Toxicol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhang, Z., Yang, H., Han, F., and Guo, P. (2025). Reactive Oxygen Species as Key Molecules in the Pathogenesis of Alcoholic Fatty Liver Disease and Nonalcoholic Fatty Liver Disease: Future Perspectives. Curr. Issues Mol. Biol., 47.","DOI":"10.3390\/cimb47060464"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"102010","DOI":"10.1016\/j.trim.2024.102010","article-title":"Protective Effects and Mechanisms of Dapagliflozin on Renal Ischemia\/Reperfusion Injury","volume":"84","author":"Li","year":"2024","journal-title":"Transpl. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"108503","DOI":"10.1016\/j.intimp.2021.108503","article-title":"Dapagliflozin Improves Steatohepatitis in Diabetic Rats via Inhibition of Oxidative Stress and Inflammation","volume":"104","author":"Hazem","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Paul, M., Hemshekhar, M., Thushara, R.M., Sundaram, M.S., NaveenKumar, S.K., Naveen, S., Devaraja, S., Somyajit, K., West, R. (2015). Methotrexate Promotes Platelet Apoptosis via JNK-Mediated Mitochondrial Damage: Alleviation by N-Acetylcysteine and N-Acetylcysteine Amide. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127558"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"45120","DOI":"10.1074\/jbc.M106386200","article-title":"Tumor Necrosis Factor-\u03b1 Induces Bax-Bak Interaction and Apoptosis, Which Is Inhibited by Adenovirus E1B 19K","volume":"276","author":"Sundararajan","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2349","DOI":"10.1101\/gad.272278.115","article-title":"Atomic Structure of the Apoptosome: Mechanism of Cytochrome c and DATP-Mediated Activation of Apaf-1","volume":"29","author":"Zhou","year":"2015","journal-title":"Genes Dev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1089\/ars.2014.5851","article-title":"Endoplasmic Reticulum Stress and Oxidative Stress in Cell Fate Decision and Human Disease","volume":"21","author":"Cao","year":"2014","journal-title":"Antioxid. Redox Signal."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/s41392-025-02176-0","article-title":"The BCL2 Family: From Apoptosis Mechanisms to New Advances in Targeted Therapy","volume":"10","author":"Vogler","year":"2025","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e129","DOI":"10.2500\/ajra.2011.25.3618","article-title":"Effects of Methotrexate on Vascular Endothelial Growth Factor, Angiopoietin 1, and Angiopoietin 2 in Nasal Polyps","volume":"25","author":"Park","year":"2011","journal-title":"Am. J. Rhinol. Allergy"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/S0002-9440(10)64525-7","article-title":"Overexpression of Vascular Endothelial Growth Factor (VEGF) in the Retinal Pigment Epithelium Leads to the Development of Choroidal Neovascularization","volume":"157","author":"Spilsbury","year":"2000","journal-title":"Am. J. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"109686","DOI":"10.1016\/j.intimp.2023.109686","article-title":"Gastroprotective Effect of Dapagliflozin in Ethanol-Induced Gastric Lesions in Rats: Crosstalk between HMGB1\/RAGE\/PTX3 and TLR4\/MyD88\/VEGF\/PDGF Signaling Pathways","volume":"115","author":"Salama","year":"2023","journal-title":"Int. Immunopharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"715","DOI":"10.17219\/acem\/68967","article-title":"Protective Effects of Tumor Necrosis Factor Alpha Inhibitors on Methotrexate-Induced Pancreatic Toxicity","volume":"27","author":"Mercantepe","year":"2018","journal-title":"Adv. Clin. Exp. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13","DOI":"10.2147\/DDDT.S181550","article-title":"Cytoprotective Effects of Molsidomine against Methotrexate-Induced Hepatotoxicity: An Experimental Rat Study","volume":"13","author":"Huz","year":"2018","journal-title":"Drug Des. Devel. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"112628","DOI":"10.1016\/j.biopha.2022.112628","article-title":"The Dynamic Interplay between AMPK\/NF\u03baB Signaling and NLRP3 Is a New Therapeutic Target in Inflammation: Emerging Role of Dapagliflozin in Overcoming Lipopolysaccharide-Mediated Lung Injury","volume":"147","author":"Saber","year":"2022","journal-title":"Biomed. Pharmacother."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1007\/s00210-020-01843-z","article-title":"Protective Effects of Hesperidin against MTX-Induced Hepatotoxicity in Male Albino Rats","volume":"393","author":"Abdelaziz","year":"2020","journal-title":"Naunyn. Schmiedebergs. Arch. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"621","DOI":"10.33549\/physiolres.935338","article-title":"Can Alpha-Pinene Prevent Methotrexate-Induced Cardiac and Hepatic Damage?","volume":"73","author":"Cirrik","year":"2024","journal-title":"Physiol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5129","DOI":"10.1007\/s00210-024-03567-w","article-title":"Dabigatran Attenuates Methotrexate-Induced Hepatotoxicity by Regulating Coagulation, Endothelial Dysfunction, and the NF-KB\/IL-1\u03b2\/MCP-1 and TLR4\/NLRP3 Signaling Pathways","volume":"398","author":"Alzokaky","year":"2025","journal-title":"Naunyn. Schmiedebergs. Arch. Pharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1080\/01480545.2024.2425994","article-title":"Cannabidiol Mitigates Methotrexate-Induced Hepatic Injury via SIRT-1\/P53 Signaling and Mitochondrial Pathways: Reduces Oxidative Stress and Inflammation","volume":"48","author":"Ilhan","year":"2025","journal-title":"Drug Chem. Toxicol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/j.clinbiochem.2003.11.015","article-title":"A Novel Automated Direct Measurement Method for Total Antioxidant Capacity Using a New Generation, More Stable ABTS Radical Cation","volume":"37","author":"Erel","year":"2004","journal-title":"Clin. Biochem."}],"updated-by":[{"DOI":"10.3390\/ijms27073158","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000}}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2110\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T13:36:08Z","timestamp":1774964168000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2110"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,24]]},"references-count":41,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["ijms27052110"],"URL":"https:\/\/doi.org\/10.3390\/ijms27052110","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,24]]}}}