{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:29:59Z","timestamp":1760059799508,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T00:00:00Z","timestamp":1752624000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Centre, Poland","doi-asserted-by":"publisher","award":["DEC-2016\/21\/D\/NZ9\/01301"],"award-info":[{"award-number":["DEC-2016\/21\/D\/NZ9\/01301"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Zearalenone (ZEN), a mycotoxin commonly present in maize-based feed, poses a health risk to dairy cows. While the hepatic effects of ZEN are increasingly studied, little is known about its impact on cows with altered energy metabolism. This study investigated the transcriptional response of liver cells isolated from healthy and ketotic cows to ZEN exposure using a novel in vitro model. Hepatocytes were obtained via biopsy from 12 cows, cultured under standardized conditions, and exposed to ZEN (0\u2013100 \u00b5M) for 1, 3, and 6 h. Gene expression analysis focused on targets related to glucose and lipid metabolism. ZEN induced time- and dose-dependent changes in gene expression, with the most prominent effects observed after 1 h. Key metabolic genes were differentially regulated depending on the cow\u2019s metabolic status. Notably, hepatocytes from healthy cows showed a stronger transcriptional response than those from ketotic cows, indicating reduced metabolic adaptability in energy-compromised animals. Significant interaction effects between ZEN dose and metabolic status were observed, especially for genes involved in glycolysis and fatty acid metabolism. This study presents a novel in vitro model and emphasizes the need to consider metabolic health when assessing the risks of mycotoxin exposure in dairy cattle.<\/jats:p>","DOI":"10.3390\/ijms26146827","type":"journal-article","created":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T15:48:22Z","timestamp":1752680902000},"page":"6827","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Differential Impact of Zearalenone on Hepatic Glucose and Lipid Metabolism in Healthy and Ketotic Dairy Cows: An In Vitro Study"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3249-4538","authenticated-orcid":false,"given":"Justyna","family":"Bar\u0107","sequence":"first","affiliation":[{"name":"Department of Animal Nutrition and Fisheries, University of Agriculture in Krakow, Al. Mickiewicza 24\/28, 30-059 Krakow, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3935-9531","authenticated-orcid":false,"given":"Zygmunt Maciej","family":"Kowalski","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Fisheries, University of Agriculture in Krakow, Al. Mickiewicza 24\/28, 30-059 Krakow, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wojciech","family":"Jagusiak","sequence":"additional","affiliation":[{"name":"Department of Animal Genetics, Breeding and Ethology, University of Agriculture in Krakow, Al. Mickiewicza 24\/28, 30-059 Krakow, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2420","DOI":"10.1016\/j.fct.2007.06.018","article-title":"Toxigenic fungi and mycotoxins in mature corn silage","volume":"45","author":"Richard","year":"2007","journal-title":"Food Chem. Toxicol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1080\/19393210802236927","article-title":"Occurrence of mycotoxins in maize, grass and wheat silage for dairy cattle in the Netherlands","volume":"1","author":"Driehuis","year":"2008","journal-title":"Food Addit. Contam. Part B"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3057","DOI":"10.3390\/toxins7083057","article-title":"Review on mycotoxin issues in ruminants: Occurrence in forages, effects of mycotoxin ingestion on health status and animal performance and practical strategies to counteract their negative effects","volume":"7","author":"Gallo","year":"2015","journal-title":"Toxins"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1016\/j.anifeedsci.2007.06.009","article-title":"Methods for preventing, decontaminating and minimizing the toxicity of mycotoxins in feeds","volume":"137","author":"Jouany","year":"2007","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1017\/S1751731114000731","article-title":"Prepartum and postpartum nutritional management to optimize fertility in high-yielding dairy cows in confined TMR systems","volume":"8","author":"Drackley","year":"2014","journal-title":"Animal"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1615","DOI":"10.2527\/1992.7051615x","article-title":"Mycotoxins and reproduction in domestic livestock","volume":"70","author":"Diekman","year":"1992","journal-title":"J. Anim. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1111\/j.1439-0442.1984.tb01327.x","article-title":"Concentrations of blood and milk ketone bodies, blood isopropanol and plasma glucose in dairy cows in relation to the degree of hyperketonemia and clinical signs","volume":"31","author":"Andersson","year":"1984","journal-title":"Zentralblatt Veterin\u00e4rmedizin Reihe A"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"571","DOI":"10.3168\/jds.2008-1507","article-title":"Impact of hyperketonemia in early lactation dairy cows on health and production","volume":"92","author":"Duffield","year":"2009","journal-title":"J. Dairy Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.cvfa.2004.06.006","article-title":"Monitoring and testing dairy herds for metabolic disease","volume":"20","author":"Oetzel","year":"2004","journal-title":"Vet. Clin. Food Anim. Pract."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"9532","DOI":"10.3168\/jds.2022-23081","article-title":"Association between hyperketolactia and production in early-lactating dairy cows","volume":"106","author":"Kowalski","year":"2023","journal-title":"J. Dairy Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1007\/PL00000897","article-title":"Cytochromes P450 and metabolism of xenobiotics","volume":"58","author":"Anzenbacher","year":"2001","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_12","first-page":"417","article-title":"Toxic effects of the mycotoxin zearalenone and its derivatives on in vitro maturation of bovine oocytes","volume":"55","author":"Minervini","year":"2001","journal-title":"Theriogenology"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Falkauskas, R., Bakutis, B., Jovai\u0161ien\u0117, J., Vai\u010diulien\u0117, G., Gerulis, G., Kerzien\u0117, S., Jacevi\u010dien\u0117, I., Jacevi\u010dius, E., and Baliukonien\u0117, V. (2022). Zearalenone and its metabolites in blood serum, urine, and milk of dairy cows. Animals, 12.","DOI":"10.3390\/ani12131651"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Gruber-Dorninger, C., Faas, J., Doupovec, B., Aleschko, M., Stoiber, C., H\u00f6bartner-Gu\u00dfl, A., Sch\u00f6ndorfer, K., Killinger, M., Zebeli, Q., and Schatzmayr, D. (2021). Metabolism of zearalenone in the rumen of dairy cows with and without application of a zearalenone-degrading enzyme. Toxins, 13.","DOI":"10.3390\/toxins13020084"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Bar\u0107, J., Flaga, J., Kozubek, A., and Kowalski, Z.M. (2023). Short culture of bovine hepatocytes biopsied from dairy cows as a model for toxicological studies\u2014CYP 1A1 activity response to zearalenone treatment. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241512344"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1152\/physiolgenomics.00188.2007","article-title":"Nutrition-induced ketosis alters metabolic and signaling gene networks in liver of periparturient dairy cows","volume":"32","author":"Loor","year":"2007","journal-title":"Physiol. Genom."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Ning, M., Zhao, Y., Li, Z., and Cao, J. (2022). Ketosis alters transcriptional adaptations of subcutaneous white adipose tissue in Holstein cows during the transition period. Animals, 12.","DOI":"10.3390\/ani12172238"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3367","DOI":"10.3168\/jds.2023-23904","article-title":"Invited review: Ketone biology\u2014The shifting paradigm of ketones and ketosis in the dairy cow","volume":"107","author":"Rico","year":"2024","journal-title":"J. Dairy Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2577","DOI":"10.1007\/s00018-013-1539-2","article-title":"Regulation of pyruvate metabolism and human disease","volume":"71","author":"Gray","year":"2014","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"602","DOI":"10.3168\/jds.2020-18883","article-title":"Differential gene expression in dairy cows under negative energy balance and ketosis: A systematic review and meta-analysis","volume":"104","author":"Soares","year":"2021","journal-title":"J. Dairy Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1648","DOI":"10.1038\/s42255-021-00489-2","article-title":"Peroxisomal \u03b2-oxidation acts as a sensor for intracellular fatty acids and regulates lipolysis","volume":"3","author":"Ding","year":"2021","journal-title":"Nat. Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"136637","DOI":"10.1016\/j.chemosphere.2022.136637","article-title":"Characterization of zearalenone-induced hepatotoxicity and its mechanisms by transcriptomics in zebrafish model","volume":"309","author":"Zhang","year":"2022","journal-title":"Chemosphere"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Cheng, Q., Jiang, S., Huang, L., Wang, Y., and Yang, W. (2022). Zearalenone exposure affects the Keap1\u2013Nrf2 signaling pathway and glucose nutrient absorption related genes of porcine jejunal epithelial cells. Toxins, 14.","DOI":"10.3390\/toxins14110793"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"117664","DOI":"10.1016\/j.ecoenv.2024.117664","article-title":"Zearalenone exacerbates lipid metabolism disorders by promoting liver lipid droplet formation and disrupting gut microbiota","volume":"289","author":"Han","year":"2025","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1038\/s41597-019-0313-1","article-title":"Transcriptome and chromatin accessibility in porcine intestinal epithelial cells upon zearalenone exposure","volume":"6","author":"Wang","year":"2019","journal-title":"Sci. Data"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Jung, Y.Y., Kim, H.M., and Koo, J.S. (2015). Expression of lipid metabolism-related proteins in metastatic breast cancer. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137204"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Yang, C., Liu, J., Wu, X., Bao, P., Long, R., Guo, X., and Yan, P. (2017). The response of gene expression associated with lipid metabolism, fat deposition and fatty acid profile in the longissimus dorsi muscle of Gannan yaks to different energy levels of diets. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0187604"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Yoon, J.E., Lee, K.Y., Seok, J.S., Cheng, W.N., Kwon, H.C., Jeong, C.H., and Han, S.G. (2019). Zearalenone induces endoplasmic reticulum stress and modulates the expression of phase I\/II enzymes in human liver cells. Toxins, 12.","DOI":"10.3390\/toxins12010002"},{"key":"ref_29","first-page":"251","article-title":"Comparative analysis on gene expression profiles in cattle subcutaneous fat tissues","volume":"3","author":"Taniguchi","year":"2008","journal-title":"Comp. Biochem. Physiol. D Genom. Proteom."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/S0278-6915(01)00118-1","article-title":"Methods of in vitro toxicology","volume":"40","author":"Eisenbrand","year":"2002","journal-title":"Food Chem. Toxicol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2618","DOI":"10.3168\/jds.2008-1795","article-title":"Evaluation of an electronic cowside test to detect subclinical ketosis in dairy cows","volume":"92","author":"Iwersen","year":"2009","journal-title":"J. Dairy Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2208","DOI":"10.3168\/jds.S0022-0302(95)76848-5","article-title":"Time trends of plasma lipids and enzymes synthesizing hepatic triacylglycerol during postpartum development of fatty liver in dairy cows","volume":"78","author":"Wensing","year":"1995","journal-title":"J. Dairy Sci."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Panda, S., Bisht, S., Malakar, D., Mohanty, A.K., and Kaushik, J.K. (2015). In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0118841"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1007\/s10616-006-9000-0","article-title":"Culture of bovine hepatocytes: A non-perfusion technique for cell isolation","volume":"51","author":"Spotorno","year":"2006","journal-title":"Cytotechnology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/j.tiv.2003.12.011","article-title":"Cytotoxicity, inhibition of DNA and protein syntheses and oxidative damage in cultured cells exposed to zearalenone","volume":"18","author":"Ouanes","year":"2004","journal-title":"Toxicol In Vitro"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1671","DOI":"10.1016\/j.tiv.2008.06.016","article-title":"The mycotoxin Zearalenone induces apoptosis in human hepatocytes (HepG2) via p53-dependent mitochondrial signaling pathway","volume":"22","author":"Bouaziz","year":"2008","journal-title":"Toxicol In Vitro"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/14\/6827\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:10:56Z","timestamp":1760033456000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/14\/6827"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,16]]},"references-count":36,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["ijms26146827"],"URL":"https:\/\/doi.org\/10.3390\/ijms26146827","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2025,7,16]]}}}