{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T03:08:04Z","timestamp":1789787284259,"version":"4.0.1"},"reference-count":50,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Postgraduate Study Support Grant of the Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand."},{"name":"Invitation Research of the Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.","award":["IN 66077"],"award-info":[{"award-number":["IN 66077"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Methotrexate (MTX) is used in treating several malignancies. However, MTX neurotoxicity remains a significant clinical side effect, leading to cell division malformation, and neurogenesis impairment. Chrysin, a flavonoid compound found in natural products, demonstrates various biological characteristics, including neuroprotective and antioxidant properties. The purpose of this study was to investigate the ameliorative effect of chrysin on oxidative damage and neurogenesis impairment caused by MTX. Male Sprague-Dawley rats were randomly divided into four groups, including the vehicle, MTX (75 mg\/kg), chrysin (10 mg\/kg), and chrysin+MTX groups. Chrysin was orally administered for 15 days. MTX was administered intravenously on days 8 and 15. The hippocampal neural stem cells were evaluated using sex determining region Y-box 2 (sox2) and nestin immunofluorescence staining. Antioxidant enzyme expression and the levels of oxidative stress marker were assessed. Additionally, the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), brain-derived neurotrophic factor (BDNF), cAMP-response element binding (CREB), and phosphorylated CREB (pCREB) were evaluated using Western blotting. Results showed that MTX significantly decreased the activity of antioxidant enzymes and produced oxidative stress. MTX also impaired neurogenesis, evidenced by decreased sox2 and nestin-positive cells and decreased expression of Nrf2, BDNF, CREB, and pCREB in the hippocampus and prefrontal cortex. However, chrysin significantly reversed the effects of MTX on these parameters. In conclusion, chrysin exhibits neuroprotective effects against MTX-induced neurogenesis impairment by upregulating antioxidant enzyme activity, reducing oxidative stress, and improving protein expression related to neurogenesis.<\/jats:p>","DOI":"10.1371\/journal.pone.0342921","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T18:30:36Z","timestamp":1771353036000},"page":"e0342921","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Chrysin ameliorates methotrexate-induced hippocampal neurogenesis impairment by suppressing of oxidative stress and upregulating antioxidant enzyme activity in rodents"],"prefix":"10.1371","volume":"21","author":[{"given":"Tanaporn","family":"Anosri","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soraya","family":"Kaewngam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ram","family":"Prajit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kornrawee","family":"Suwannakot","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nataya","family":"Sritawan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anusara","family":"Aranarochana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanassanan","family":"Pannangrong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jariya Umka","family":"Welbat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Wigmore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6929-7709","authenticated-orcid":true,"given":"Apiwat","family":"Sirichoat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,2,17]]},"reference":[{"issue":"4","key":"pone.0342921.ref001","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1007\/s10495-005-2947-z","article-title":"Ornithine decarboxylase prevents methotrexate-induced apoptosis by reducing intracellular reactive oxygen species production","volume":"10","author":"C-C Huang","year":"2005","journal-title":"Apoptosis"},{"issue":"5","key":"pone.0342921.ref002","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1002\/art.20278","article-title":"Toward a better understanding of methotrexate","volume":"50","author":"JM Kremer","year":"2004","journal-title":"Arthritis Rheum"},{"issue":"1","key":"pone.0342921.ref003","doi-asserted-by":"crossref","first-page":"18548","DOI":"10.1038\/s41598-024-69263-0","article-title":"Potential neurotoxicity associated with methotrexate","volume":"14","author":"N Apiraksattayakul","year":"2024","journal-title":"Sci Rep"},{"key":"pone.0342921.ref004","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.bcp.2019.02.010","article-title":"Melatonin protects against methotrexate-induced memory deficit and hippocampal neurogenesis impairment in a rat model","volume":"163","author":"A Sirichoat","year":"2019","journal-title":"Biochem Pharmacol"},{"key":"pone.0342921.ref005","doi-asserted-by":"crossref","first-page":"110651","DOI":"10.1016\/j.biopha.2020.110651","article-title":"Metformin alleviates memory and hippocampal neurogenesis decline induced by methotrexate chemotherapy in a rat model","volume":"131","author":"N Sritawan","year":"2020","journal-title":"Biomed Pharmacother"},{"issue":"4","key":"pone.0342921.ref006","doi-asserted-by":"crossref","first-page":"936","DOI":"10.3390\/nu11040936","article-title":"Hesperidin Alleviates Methotrexate-Induced Memory Deficits via Hippocampal Neurogenesis in Adult Rats","volume":"11","author":"S Naewla","year":"2019","journal-title":"Nutrients"},{"issue":"3","key":"pone.0342921.ref007","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1523\/JNEUROSCI.22-03-00612.2002","article-title":"Neurogenesis in the adult brain","volume":"22","author":"FH Gage","year":"2002","journal-title":"J Neurosci"},{"issue":"22","key":"pone.0342921.ref008","doi-asserted-by":"crossref","first-page":"3385","DOI":"10.1007\/s00213-017-4720-8","article-title":"Saikosaponin D relieves unpredictable chronic mild stress induced depressive-like behavior in rats: involvement of HPA axis and hippocampal neurogenesis","volume":"234","author":"H-Y Li","year":"2017","journal-title":"Psychopharmacology (Berl)"},{"key":"pone.0342921.ref009","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1159\/000082134","article-title":"SOX2, a persistent marker for multipotential neural stem cells derived from embryonic stem cells, the embryo or the adult","volume":"26","author":"P Ellis","year":"2004","journal-title":"Dev Neurosci"},{"issue":"8","key":"pone.0342921.ref010","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1369\/jhc.2010.955609","article-title":"The neural stem\/progenitor cell marker nestin is expressed in proliferative endothelial cells, but not in mature vasculature","volume":"58","author":"S Suzuki","year":"2010","journal-title":"J Histochem Cytochem"},{"issue":"11","key":"pone.0342921.ref011","doi-asserted-by":"crossref","first-page":"20913","DOI":"10.3390\/ijms151120913","article-title":"Chrysin protects against focal cerebral ischemia\/reperfusion injury in mice through attenuation of oxidative stress and inflammation","volume":"15","author":"Y Yao","year":"2014","journal-title":"Int J Mol Sci"},{"issue":"3","key":"pone.0342921.ref012","doi-asserted-by":"crossref","first-page":"2802","DOI":"10.1016\/j.foodchem.2013.05.102","article-title":"Buckwheat honeys: screening of composition and properties","volume":"141","author":"F Pasini","year":"2013","journal-title":"Food Chem"},{"key":"pone.0342921.ref013","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.phytochem.2017.09.016","article-title":"Chrysin: Sources, beneficial pharmacological activities, and molecular mechanism of action","volume":"145","author":"R Mani","year":"2018","journal-title":"Phytochemistry"},{"issue":"3","key":"pone.0342921.ref014","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1007\/s11418-014-0840-1","article-title":"Evaluation of the neuroprotective effect of chrysin via modulation of endogenous biomarkers in a rat model of spinal cord injury","volume":"68","author":"AD Kandhare","year":"2014","journal-title":"J Nat Med"},{"key":"pone.0342921.ref015","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.ejphar.2012.01.025","article-title":"Chrysin improves cognitive deficits and brain damage induced by chronic cerebral hypoperfusion in rats","volume":"680","author":"X-L He","year":"2012","journal-title":"Eur J Pharmacol"},{"issue":"6","key":"pone.0342921.ref016","doi-asserted-by":"crossref","first-page":"1607","DOI":"10.1007\/s00213-020-05482-3","article-title":"Chrysin attenuates traumatic brain injury-induced recognition memory decline, and anxiety\/depression-like behaviors in rats: Insights into underlying mechanisms","volume":"237","author":"M Rashno","year":"2020","journal-title":"Psychopharmacology (Berl)"},{"issue":"2","key":"pone.0342921.ref017","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1007\/s11011-019-00527-9","article-title":"Protective effects of Chrysin against memory impairment, cerebral hyperemia and oxidative stress after cerebral hypoperfusion and reperfusion in rats","volume":"35","author":"MK Shooshtari","year":"2020","journal-title":"Metab Brain Dis"},{"issue":"4","key":"pone.0342921.ref018","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.3390\/nu12041100","article-title":"Chrysin Protects against Memory and Hippocampal Neurogenesis Depletion in D-Galactose-Induced Aging in Rats","volume":"12","author":"R Prajit","year":"2020","journal-title":"Nutrients"},{"key":"pone.0342921.ref019","doi-asserted-by":"crossref","first-page":"112280","DOI":"10.1016\/j.biopha.2021.112280","article-title":"Effect of metformin treatment on memory and hippocampal neurogenesis decline correlated with oxidative stress induced by methotrexate in rats","volume":"144","author":"N Sritawan","year":"2021","journal-title":"Biomed Pharmacother"},{"key":"pone.0342921.ref020","doi-asserted-by":"crossref","first-page":"114083","DOI":"10.1016\/j.bcp.2020.114083","article-title":"Neuroprotective effects of hesperidin against methotrexate-induced changes in neurogenesis and oxidative stress in the adult rat","volume":"178","author":"JU Welbat","year":"2020","journal-title":"Biochem Pharmacol"},{"key":"pone.0342921.ref021","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.neuro.2022.06.010","article-title":"Neuroprotective properties of chrysin on decreases of cell proliferation, immature neurons and neuronal cell survival in the hippocampal dentate gyrus associated with cognition induced by methotrexate","volume":"92","author":"A Sirichoat","year":"2022","journal-title":"Neurotoxicology"},{"key":"pone.0342921.ref022","doi-asserted-by":"crossref","first-page":"1596362","DOI":"10.1155\/2022\/1596362","article-title":"Melatonin Attenuates Methotrexate-Induced Reduction of Antioxidant Activity Related to Decreases of Neurogenesis in Adult Rat Hippocampus and Prefrontal Cortex","volume":"2022","author":"K Suwannakot","year":"2022","journal-title":"Oxid Med Cell Longev"},{"issue":"6","key":"pone.0342921.ref023","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.jpha.2019.06.001","article-title":"Analytical methodologies for determination of methotrexate and its metabolites in pharmaceutical, biological and environmental samples","volume":"9","author":"F Karami","year":"2019","journal-title":"J Pharm Anal"},{"issue":"1","key":"pone.0342921.ref024","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.bcp.2011.03.020","article-title":"Neurotoxicity of methotrexate to hippocampal cells in vivo and in vitro","volume":"82","author":"M Yang","year":"2011","journal-title":"Biochem Pharmacol"},{"issue":"3","key":"pone.0342921.ref025","doi-asserted-by":"crossref","first-page":"305","DOI":"10.4252\/wjsc.v6.i3.305","article-title":"Sox2, a key factor in the regulation of pluripotency and neural differentiation","volume":"6","author":"S Zhang","year":"2014","journal-title":"World J Stem Cells"},{"key":"pone.0342921.ref026","doi-asserted-by":"crossref","first-page":"114613","DOI":"10.1016\/j.biopha.2023.114613","article-title":"Protective effect of metformin on methotrexate induced reduction of rat hippocampal neural stem cells and neurogenesis","volume":"162","author":"N Sritawan","year":"2023","journal-title":"Biomed Pharmacother"},{"key":"pone.0342921.ref027","doi-asserted-by":"crossref","first-page":"115329","DOI":"10.1016\/j.biopha.2023.115329","article-title":"Hesperidin ameliorates impairment in hippocampal neural stem cells related to apoptosis induced by methotrexate in adult rats","volume":"166","author":"S Naewla","year":"2023","journal-title":"Biomed Pharmacother"},{"issue":"9","key":"pone.0342921.ref028","doi-asserted-by":"crossref","first-page":"852","DOI":"10.3390\/antiox9090852","article-title":"Oxidative Stress: Concept and Some Practical Aspects","volume":"9","author":"H Sies","year":"2020","journal-title":"Antioxidants (Basel)"},{"issue":"8","key":"pone.0342921.ref029","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1038\/sj.cdd.4401458","article-title":"Redox regulation in disease and ageing","volume":"11","author":"M Maccarrone","year":"2004","journal-title":"Cell Death Differ"},{"issue":"2","key":"pone.0342921.ref030","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.conb.2009.05.001","article-title":"Redox dysregulation, neurodevelopment, and schizophrenia","volume":"19","author":"KQ Do","year":"2009","journal-title":"Curr Opin Neurobiol"},{"issue":"5","key":"pone.0342921.ref031","doi-asserted-by":"crossref","first-page":"439","DOI":"10.3390\/antiox9050439","article-title":"A bibliometric review of publications on oxidative stress and chemobrain: 1990-2019","volume":"9","author":"T McElroy","year":"2020","journal-title":"Antioxidants"},{"key":"pone.0342921.ref032","first-page":"453","article-title":"Malondialdehyde (MDA) as a lipid peroxidation marker","volume":"57","author":"S Gawe\u0142","year":"2004","journal-title":"Wiad Lek"},{"issue":"12","key":"pone.0342921.ref033","doi-asserted-by":"crossref","first-page":"2345","DOI":"10.3390\/antiox11122345","article-title":"Nrf2 and Oxidative Stress: A General Overview of Mechanisms and Implications in Human Disease","volume":"11","author":"V Ngo","year":"2022","journal-title":"Antioxidants (Basel)"},{"issue":"13","key":"pone.0342921.ref034","doi-asserted-by":"crossref","first-page":"4777","DOI":"10.3390\/ijms21134777","article-title":"NRF2, a Transcription Factor for Stress Response and Beyond","volume":"21","author":"F He","year":"2020","journal-title":"Int J Mol Sci"},{"key":"pone.0342921.ref035","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.lfs.2019.04.027","article-title":"Chrysin prevents cognitive and hippocampal long-term potentiation deficits and inflammation in rat with cerebral hypoperfusion and reperfusion injury","volume":"226","author":"A Sarkaki","year":"2019","journal-title":"Life Sci"},{"key":"pone.0342921.ref036","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.pbb.2015.04.010","article-title":"Flavonoid Chrysin prevents age-related cognitive decline via attenuation of oxidative stress and modulation of BDNF levels in aged mouse brain","volume":"134","author":"LC Souza","year":"2015","journal-title":"Pharmacol Biochem Behav"},{"issue":"2","key":"pone.0342921.ref037","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.abb.2008.04.003","article-title":"(-)-Epigallocatechin gallate induces Nrf2-mediated antioxidant enzyme expression via activation of PI3K and ERK in human mammary epithelial cells","volume":"476","author":"H-K Na","year":"2008","journal-title":"Arch Biochem Biophys"},{"key":"pone.0342921.ref038","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/j.neuroscience.2014.12.048","article-title":"Chronic unpredictable mild stress decreases BDNF and NGF levels and Na(+),K(+)-ATPase activity in the hippocampus and prefrontal cortex of mice: antidepressant effect of chrysin","volume":"289","author":"CB Filho","year":"2015","journal-title":"Neuroscience"},{"issue":"6153","key":"pone.0342921.ref039","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1038\/331261a0","article-title":"Brain-derived neurotrophic factor prevents neuronal death in vivo","volume":"331","author":"MM Hofer","year":"1988","journal-title":"Nature"},{"key":"pone.0342921.ref040","doi-asserted-by":"crossref","first-page":"26","DOI":"10.3389\/fnins.2016.00026","article-title":"Regulation of Neurogenesis by Neurotrophins during Adulthood: Expected and Unexpected Roles","volume":"10","author":"M Vilar","year":"2016","journal-title":"Front Neurosci"},{"issue":"6","key":"pone.0342921.ref041","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.5114\/aoms.2015.56342","article-title":"Brain-derived neurotrophic factor and its clinical implications","volume":"11","author":"S Bathina","year":"2015","journal-title":"Arch Med Sci"},{"issue":"1","key":"pone.0342921.ref042","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/ng882","article-title":"Disruption of CREB function in brain leads to neurodegeneration","volume":"31","author":"T Mantamadiotis","year":"2002","journal-title":"Nat Genet"},{"issue":"4","key":"pone.0342921.ref043","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/j.mcn.2008.07.019","article-title":"Regulation of neural migration by the CREB\/CREM transcription factors and altered Dab1 levels in CREB\/CREM mutants","volume":"39","author":"C D\u00edaz-Ruiz","year":"2008","journal-title":"Mol Cell Neurosci"},{"issue":"7","key":"pone.0342921.ref044","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1523\/JNEUROSCI.0367-19.2019","article-title":"CREB Family Transcription Factors Are Major Mediators of BDNF Transcriptional Autoregulation in Cortical Neurons","volume":"40","author":"E-E Esvald","year":"2020","journal-title":"J Neurosci"},{"issue":"20","key":"pone.0342921.ref045","doi-asserted-by":"crossref","first-page":"11024","DOI":"10.1073\/pnas.191352298","article-title":"Calcium regulation of neuronal gene expression","volume":"98","author":"AE West","year":"2001","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"4","key":"pone.0342921.ref046","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/S0896-6273(00)81010-7","article-title":"Ca2+ influx regulates BDNF transcription by a CREB family transcription factor-dependent mechanism","volume":"20","author":"X Tao","year":"1998","journal-title":"Neuron"},{"issue":"3","key":"pone.0342921.ref047","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.3892\/mmr.2016.5470","article-title":"Increased protein expression levels of pCREB, BDNF and SDF-1\/CXCR4 in the hippocampus may be associated with enhanced neurogenesis induced by environmental enrichment","volume":"14","author":"XQ Zhang","year":"2016","journal-title":"Mol Med Rep"},{"issue":"35","key":"pone.0342921.ref048","doi-asserted-by":"crossref","first-page":"27986","DOI":"10.1039\/C4RA13315C","article-title":"Free radicals, natural antioxidants, and their reaction mechanisms","volume":"5","author":"SB Nimse","year":"2015","journal-title":"RSC Adv"},{"key":"pone.0342921.ref049","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/j.neuroscience.2014.12.048","article-title":"Chronic unpredictable mild stress decreases BDNF and NGF levels and Na(+),K(+)-ATPase activity in the hippocampus and prefrontal cortex of mice: antidepressant effect of chrysin","volume":"289","author":"CB Filho","year":"2015","journal-title":"Neuroscience"},{"issue":"11","key":"pone.0342921.ref050","doi-asserted-by":"crossref","first-page":"3551","DOI":"10.3390\/molecules27113551","article-title":"Pharmacological, Neurochemical, and Behavioral Mechanisms Underlying the Anxiolytic- and Antidepressant-like Effects of Flavonoid Chrysin","volume":"27","author":"JF Rodr\u00edguez-Landa","year":"2022","journal-title":"Molecules"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342921","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T18:30:44Z","timestamp":1771353044000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342921"}},"subtitle":[],"editor":[{"given":"Stephen D.","family":"Ginsberg","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,2,17]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2,17]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0342921","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,17]]}}}