{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T05:18:22Z","timestamp":1783401502064,"version":"3.54.6"},"reference-count":26,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T00:00:00Z","timestamp":1783123200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Vitamin K is a well-established cofactor for \u03b3-glutamyl carboxylase in vertebrates, yet its physiological functions in insects remain poorly understood. Warfarin, a vitamin K antagonist, inhibits vitamin K epoxide reductase, thereby blocking the vitamin K cycle. We have previously demonstrated that, in Drosophila melanogaster, vitamin K3 (menadione sodium bisulfite, MSB) is converted into endogenous vitamin K2, and that warfarin interferes with this cycle, leading to reduced oxidative stress. In the present study, we investigated the effects of chronic exposure to MSB (3.5 mM) and\/or warfarin (1 mM and 10 mM) during larval development on adult survival, lifespan, mitochondrial ATP hydrolysis rate, and the transcriptional expression of target genes. Warfarin at 1 mM did not affect development or egg laying, whereas 10 mM caused significant postembryonic lethality. Transcriptional analysis revealed marked sex-specific effects. In females, warfarin downregulated the levy gene (encoding a complex IV subunit) and the ATPsynB gene (encoding an ATP synthase subunit B), while MSB suppressed cell cycle genes (cmet, sti, mcm2). In contrast, males exhibited upregulation of COQ7 (encoding a key enzyme in ubiquinone biosynthesis) in response to warfarin and increased sti expression in response to MSB, suggesting a more robust compensatory response. Notably, the combined application of MSB and warfarin increased mitochondrial ATP hydrolysis, thereby offsetting the negative physiological effects despite persistent transcriptional suppression. In females, either MSB or warfarin alone significantly reduced median lifespan; however, when applied together, female lifespan was restored to control levels. In males, the effects were considerably weaker. Thus, this study reveals a novel, sex-specific interaction between vitamin K and its antagonist in D. melanogaster, providing new insights into mitochondrial adaptation and sexual dimorphism in metabolic stress responses.<\/jats:p>","DOI":"10.3390\/ijms27136026","type":"journal-article","created":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T13:43:14Z","timestamp":1783345394000},"page":"6026","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Sex-Related Effect of Chronic Doses of Warfarin and Menadione on Drosophila melanogaster"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-7217-6667","authenticated-orcid":false,"given":"Anna","family":"Lavrenova","sequence":"first","affiliation":[{"name":"Faculty of Biology, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Kozlova","sequence":"additional","affiliation":[{"name":"Faculty of Biology, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0383-9237","authenticated-orcid":false,"given":"Oleg","family":"Klychnikov","sequence":"additional","affiliation":[{"name":"Faculty of Biology, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lidia","family":"Nefedova","sequence":"additional","affiliation":[{"name":"Faculty of Biology, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bus, K., and Szterk, A. (2021). Relationship between Structure and Biological Activity of Various Vitamin K Forms. Foods, 10.","DOI":"10.3390\/foods10123136"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7001","DOI":"10.3390\/cimb46070418","article-title":"Beyond the Coagulation Cascade: Vitamin K and Its Multifaceted Impact on Human and Domesticated Animal Health","volume":"46","author":"Sadler","year":"2024","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1873","DOI":"10.1111\/j.1538-7836.2005.01419.x","article-title":"The Vitamin K Cycle","volume":"3","author":"Stafford","year":"2005","journal-title":"J. Thromb. Haemost."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Majiene, D., Kuseliauskyte, J., Stimbirys, A., and Jekabsone, A. (2019). Comparison of the Effect of Native 1,4-Naphthoquinones Plumbagin, Menadione, and Lawsone on Viability, Redox Status, and Mitochondrial Functions of C6 Glioblastoma Cells. Nutrients, 11.","DOI":"10.3390\/nu11061294"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/j.taap.2012.04.030","article-title":"Rapid Generation of Mitochondrial Superoxide Induces Mitochondrion-Dependent but Caspase-Independent Cell Death in Hippocampal Neuronal Cells That Morphologically Resembles Necroptosis","volume":"262","author":"Fukui","year":"2012","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3635","DOI":"10.1016\/0006-2952(82)90587-1","article-title":"A Comparison of Vitamin K Antagonism by Warfarin, Difenacoum and Brodifacoum in the Rabbit","volume":"31","author":"Park","year":"1982","journal-title":"Biochem. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1038\/nsmb.3338","article-title":"Warfarin and Vitamin K Compete for Binding to Phe55 in Human VKOR","volume":"24","author":"Czogalla","year":"2017","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1111\/j.1538-7836.2009.03630.x","article-title":"Warfarin Use and the Risk of Valvular Calcification","volume":"7","author":"Lerner","year":"2009","journal-title":"J. Thromb. Haemost."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1182\/blood-2018-01-830901","article-title":"Warfarin and Vitamin K Epoxide Reductase: A Molecular Accounting for Observed Inhibition","volume":"132","author":"Wu","year":"2018","journal-title":"Blood"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"490","DOI":"10.5858\/2009-0474-OA.1","article-title":"International Normalized Ratio Versus Plasma Levels of Coagulation Factors in Patients on Vitamin K Antagonist Therapy","volume":"135","author":"Gulati","year":"2011","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2825","DOI":"10.1111\/jth.15519","article-title":"Overall Survival with Warfarin vs. Low-molecular-weight Heparin in Cancer-associated Thrombosis","volume":"19","author":"Chiasakul","year":"2021","journal-title":"J. Thromb. Haemost."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11748","DOI":"10.1038\/s41598-025-96681-5","article-title":"Warfarin Use and Vestibular Dysfunction Insights from NHANES Data, Network Pharmacology, Mendelian Randomization, and Molecular Docking","volume":"15","author":"Liu","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Lavrenova, A., Klychnikov, O., Ioutsi, V., Rodin, I., Luneva, O., and Nefedova, L. (2025). Effect of Warfarin on Lifespan and Oxidative Stress Tolerance of Drosophila melanogaster. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26104808"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1038\/s41586-023-06271-6","article-title":"A Rise-to-Threshold Process for a Relative-Value Decision","volume":"619","author":"Vijayan","year":"2023","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Be\u021biu, A.M., Noveanu, L., H\u00e2ncu, I.M., Lascu, A., Petrescu, L., Maack, C., Elm\u00e9r, E., and Muntean, D.M. (2022). Mitochondrial Effects of Common Cardiovascular Medications: The Good, the Bad and the Mixed. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113653"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Aghdam, M.N., Unselt, D., Adonay, M.E., Morozova, T.V., Carbone, M.A., Arya, G.H., Turlapati, L., Shankar, V., Anholt, R.R.H., and Mackay, T.F.C. (2025). Systems Genetics of Lifespan and Senescence in Drosophila melanogaster. BMC Biol., 24.","DOI":"10.1186\/s12915-025-02485-4"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1016\/j.cub.2009.08.016","article-title":"Extension of Drosophila Life Span by RNAi of the Mitochondrial Respiratory Chain","volume":"19","author":"Copeland","year":"2009","journal-title":"Curr. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Morrow, G., and Tanguay, R.M. (2015). Drosophila melanogaster Hsp22: A Mitochondrial Small Heat Shock Protein Influencing the Aging Process. Front. Genet., 6.","DOI":"10.3389\/fgene.2015.00103"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1016\/j.cmet.2016.03.009","article-title":"Mitochondrial ROS Produced via Reverse Electron Transport Extend Animal Lifespan","volume":"23","author":"Sriram","year":"2016","journal-title":"Cell Metab."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.cophys.2018.04.002","article-title":"Sex Differences in Drosophila Development and Physiology","volume":"6","author":"Millington","year":"2018","journal-title":"Curr. Opin. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Lai, Y., Loy, F., Isola, M., Noli, R., Rinaldi, A., Lobina, C., Vargiu, R., Cesare Marincola, F., and Isola, R. (2023). Male and Female Mitochondria Respond Differently after Exercising in Acute Hypoxia. Biomedicines, 11.","DOI":"10.3390\/biomedicines11123149"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.1016\/j.cmet.2016.05.019","article-title":"Sex Differences in Lifespan","volume":"23","author":"Austad","year":"2016","journal-title":"Cell Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s13293-020-00308-5","article-title":"Sex Differences in Pharmacokinetics Predict Adverse Drug Reactions in Women","volume":"11","author":"Zucker","year":"2020","journal-title":"Biol. Sex Differ."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"56147","DOI":"10.18632\/oncotarget.11269","article-title":"OASIS 2: Online Application for Survival Analysis 2 with Features for the Analysis of Maximal Lifespan and Healthspan in Aging Research","volume":"7","author":"Han","year":"2016","journal-title":"Oncotarget"},{"key":"ref_25","first-page":"e5180","article-title":"Isolation of Intact Mitochondria From Drosophila melanogaster and Assessment of Mitochondrial Respiratory Capacity Using Seahorse Analyzer","volume":"15","author":"Groen","year":"2025","journal-title":"Bio Protoc."},{"key":"ref_26","first-page":"742","article-title":"Interaction of Venturicidin and Fo\u00b7F1-ATPase\/ATP Synthase of Tightly Coupled Subbacterial Particles of Paracoccus Denitrificans in Energized Membranes","volume":"87","author":"Zharova","year":"2022","journal-title":"Biochemistry"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/6026\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T04:55:19Z","timestamp":1783400119000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/6026"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,4]]},"references-count":26,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["ijms27136026"],"URL":"https:\/\/doi.org\/10.3390\/ijms27136026","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,4]]}}}