{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T10:45:40Z","timestamp":1782729940990,"version":"3.54.5"},"reference-count":70,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T00:00:00Z","timestamp":1782432000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Longjiang Technology Talent \u201cChunyan\u201d Support Program Youth Team","award":["CYQN24048"],"award-info":[{"award-number":["CYQN24048"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Background\/Objectives: Exposure to high and sustained levels of non-esterified fatty acids (NEFA) in the peripartal period is the main cause of fatty liver disease in dairy cows. Rumen-protected choline is often fed as part of the nutritional management of peripartal cows, with in vivo and in vitro data indicating positive effects of this nutrient on alleviating liver lipid accumulation. Although hepatic molecular mechanisms associated with choline supply have been studied using a target gene, protein, or metabolite approach, application of high-throughput technologies could vastly enhance fundamental knowledge on the functional role of choline. The main objective was to challenge isolated hepatocytes with a mixture of NEFA and determine proteome- and metabolome-wide effects in response to choline supply. Methods: Three healthy female calves (1 d old, 30\u201345 kg) were sacrificed to harvest hepatocytes. During a 12 h incubation, isolated hepatocytes were challenged without NEFA (control), 1.2 mM NEFA (c9-18:1, 18:2, 16:0, 18:0, and c9-16:1 at 43.5%, 4.9%, 31.9%, 14.4%, and 5.3% of total NEFA, respectively), or NEFA for 6 h followed by 10 \u03bcM choline chloride for another 6 h (NEFA + Chol). iTRAQ labeling-based protein profiling and GC\/MS-based metabolomics profiling were used to determine changes in proteins and metabolites. Differentially abundant proteins for each group comparison were determined at a threshold of 1.4-fold change. Differences in metabolite profiles were assessed via pairwise comparisons. A subset of differentially abundant proteins was validated via qRT-PCR and Western blotting. Results: Compared with the control, there were 90 proteins and 22 metabolites in the NEFA group, and 83 proteins and 29 metabolites in the NEFA + Chol. Compared with NEFA, there were 49 proteins and 17 metabolites in the NEFA + Chol group. Greater abundance of hexokinase-1 (HK1), fructose-bisphosphate aldolase (ALDOA), mitochondrial pyruvate carrier 1 (MPC1), and increased concentrations of lactate with high NEFA treatment alone suggested greater glycolytic and TCA cycle activity. Accumulation of triacylglycerol in the NEFA group was associated with lipotoxicity and markers of inflammation, such as greater abundance of prostaglandin reductase 1 (PTGR1), serious cell autophagy processes, such as greater abundance of cell division cycle 42 (CDC42), and NF\u03baB-related proteins. Choline supplementation reduced TAG partly due to greater VLDL secretion driven by greater abundance of diacylglycerol acyltransferase (DGAT1), perilipin 3 (PLIN3), and apolipoprotein C-III (APOC3). In addition, a greater abundance of carnitine O-palmitoyltransferase 1b (CPT1B) with choline suggested enhanced mitochondrial \u03b2-oxidation. Activation of the CDC42\/JNK pathway and ROS\/NF\u03baB axis-related proteins, along with depressed PI3K\/AKT\/RAC-related proteins, indicated enhanced mitochondrial autophagy in response to NEFA. Conclusions: Overall, data confirmed published effects of choline on TAG accumulation, VLDL secretion, and fatty acid oxidation, while highlighting negative effects of NEFA on the respiratory electron transport chain, autophagy, and inflammatory processes.<\/jats:p>","DOI":"10.3390\/metabo16070451","type":"journal-article","created":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T08:45:20Z","timestamp":1782722720000},"page":"451","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Proteomics and Metabolomics Reveal Novel Impacts of Choline Supply on Calf Hepatocytes Experiencing Accumulation During a Fatty Acid Challenge"],"prefix":"10.3390","volume":"16","author":[{"given":"Yaqi","family":"Chang","sequence":"first","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Jia","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaran","family":"Si","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zexin","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiachen","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhao","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhui","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1586-4365","authenticated-orcid":false,"given":"Juan J.","family":"Loor","sequence":"additional","affiliation":[{"name":"Department of Animal Sciences, Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1869-9652","authenticated-orcid":false,"given":"Bingbing","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3440-2190","authenticated-orcid":false,"given":"Wei","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Xinyang Rd. 5, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2409","DOI":"10.3168\/jds.2011-4604","article-title":"Liver protein expression in dairy cows with high liver triglycerides in early lactation","volume":"95","author":"Sejersen","year":"2012","journal-title":"J. Dairy Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"555","DOI":"10.2478\/jvetres-2019-0059","article-title":"Effects of Ketosis in Dairy Cows on Blood Biochemical Parameters, Milk Yield and Composition, and Digestive Capacity","volume":"63","author":"Yang","year":"2019","journal-title":"J. Vet. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3588","DOI":"10.3168\/jds.2021-21140","article-title":"Role of diacylglycerol O-acyltransferase (DGAT) isoforms in bovine hepatic fatty acid metabolism","volume":"105","author":"Yang","year":"2022","journal-title":"J. Dairy Sci."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Wang, F., Guo, Y., Su, X., and Cao, J. (2025). Choline as a Modulator of Periparturient Diseases in Dairy Cows. Vet. Sci., 12.","DOI":"10.3390\/vetsci12101016"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"7630","DOI":"10.3168\/jds.2022-23182","article-title":"Rumen-protected choline reduces hepatic lipidosis by increasing hepatic triacylglycerol-rich lipoprotein secretion in dairy cows","volume":"106","author":"Arshad","year":"2023","journal-title":"J. Dairy Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2413","DOI":"10.3168\/jds.2006-028","article-title":"Supplemental choline for prevention and alleviation of fatty liver in dairy cattle","volume":"90","author":"Cooke","year":"2007","journal-title":"J. Dairy Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4016","DOI":"10.3168\/jds.2011-4233","article-title":"Effect of rumen-protected choline on performance, blood metabolites, and hepatic triacylglycerols of periparturient dairy cattle","volume":"94","author":"Zom","year":"2011","journal-title":"J. Dairy Sci."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Humer, E., Bruggeman, G., and Zebeli, Q. (2019). A Meta-Analysis on the Impact of the Supplementation of Rumen-Protected Choline on the Metabolic Health and Performance of Dairy Cattle. Animals, 9.","DOI":"10.3390\/ani9080566"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"282","DOI":"10.3168\/jds.2019-16842","article-title":"Meta-analysis of the effects of supplemental rumen-protected choline during the transition period on performance and health of parous dairy cows","volume":"103","author":"Arshad","year":"2020","journal-title":"J. Dairy Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3422","DOI":"10.3168\/jds.2006-811","article-title":"Dietary L-carnitine affects periparturient nutrient metabolism and lactation in multiparous cows","volume":"90","author":"Carlson","year":"2007","journal-title":"J. Dairy Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.3168\/jds.2012-5396","article-title":"Effect of rumen-protected choline supplementation on liver and adipose gene expression during the transition period in dairy cattle","volume":"96","author":"Goselink","year":"2013","journal-title":"J. Dairy Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3296","DOI":"10.1002\/jcp.25773","article-title":"Acetoacetate induces hepatocytes apoptosis by the ROS-mediated MAPKs pathway in ketotic cows","volume":"232","author":"Du","year":"2017","journal-title":"J. Cell Physiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1328","DOI":"10.3168\/jds.2013-6654","article-title":"Effects of nonesterified fatty acids on the synthesis and assembly of very low density lipoprotein in bovine hepatocytes in vitro","volume":"97","author":"Liu","year":"2014","journal-title":"J. Dairy Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7785","DOI":"10.1038\/s41598-026-38923-8","article-title":"Modulation of fat mobilization and adipose tissue gene expression in Holstein cows supplemented with omega-3 fatty acids and N-acetyl-tryptophan during the transition period","volume":"16","author":"Ghorbanalinia","year":"2026","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"106813","DOI":"10.1016\/j.jsbmb.2025.106813","article-title":"Non-esterified fatty acids disrupt hepatic lipid metabolism and mitochondrial function via TLR4\/MyD88\/IRAK2 signaling in bovine hepatocytes","volume":"253","author":"Wen","year":"2025","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Xing, D., Wang, B., Lu, H., Peng, T., Su, J., Lei, H., He, J., Zhou, Y., and Liu, L. (2021). Sirtuin 3 Restores Synthesis and Secretion of Very Low-Density Lipoproteins in Cow Hepatocytes Challenged with Nonesterified Fatty Acids In Vitro. Vet. Sci., 8.","DOI":"10.3390\/vetsci8070121"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s13059-017-1187-1","article-title":"Dietary restriction protects from age-associated DNA methylation and induces epigenetic reprogramming of lipid metabolism","volume":"18","author":"Hahn","year":"2017","journal-title":"Genome Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2200","DOI":"10.3168\/jds.2019-17378","article-title":"Effect of prepartum energy intake and supplementation with ruminally protected choline on innate and adaptive immunity of multiparous Holstein cows","volume":"103","author":"Zenobi","year":"2020","journal-title":"J. Dairy Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"9897","DOI":"10.3168\/jds.2024-24738","article-title":"Diacylglycerol O-acyltransferase isoforms play a role in peridroplet mitochondrial fatty acid metabolism in bovine liver","volume":"107","author":"Wang","year":"2024","journal-title":"J. Dairy Sci."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lee, N.C.W., Carella, M.A., Papa, S., and Bubici, C. (2018). High Expression of Glycolytic Genes in Cirrhosis Correlates with the Risk of Developing Liver Cancer. Front. Cell Dev. Biol., 6.","DOI":"10.3389\/fcell.2018.00138"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3215","DOI":"10.1007\/s10620-019-05642-2","article-title":"Fructose-Bisphosphate Aldolase A Regulates Hypoxic Adaptation in Hepatocellular Carcinoma and Involved with Tumor Malignancy","volume":"64","author":"Li","year":"2019","journal-title":"Dig. Dis. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1002\/bdd.2329","article-title":"Involvement of SLC16A1\/MCT1 and SLC16A3\/MCT4 in l-lactate transport in the hepatocellular carcinoma cell line","volume":"43","author":"Mukai","year":"2022","journal-title":"Biopharm. Drug Dispos."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6232","DOI":"10.1021\/bi400838t","article-title":"Structural basis for regulation of human glucokinase by glucokinase regulatory protein","volume":"52","author":"Beck","year":"2013","journal-title":"Biochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"37125","DOI":"10.1074\/jbc.274.52.37125","article-title":"Nuclear import of hepatic glucokinase depends upon glucokinase regulatory protein, whereas export is due to a nuclear export signal sequence in glucokinase","volume":"274","author":"Shiota","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6700","DOI":"10.1038\/s41467-025-61939-z","article-title":"Structure of human mitochondrial pyruvate carrier MPC1 and MPC2 complex","volume":"16","author":"Sun","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"iyae058","DOI":"10.1093\/genetics\/iyae058","article-title":"Identification of factors limiting the allotopic production of the Cox2 subunit of yeast cytochrome c oxidase","volume":"227","author":"Hamel","year":"2024","journal-title":"Genetics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8713","DOI":"10.3390\/ijms15058713","article-title":"Role of mitochondria in nonalcoholic fatty liver disease","volume":"15","author":"Nassir","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"972","DOI":"10.3168\/jds.2019-17135","article-title":"Effects of abomasal infusion of essential fatty acids and conjugated linoleic acid on performance and fatty acid, antioxidative, and inflammatory status in dairy cows","volume":"103","author":"Haubold","year":"2020","journal-title":"J. Dairy Sci."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Shi, Z., Li, X.B., Peng, Z.C., Fu, S.P., Zhao, C.X., Du, X.L., Fang, Z.Y., Wang, Z., Liu, G.W., and Li, X.W. (2018). Berberine Protects against NEFA-Induced Impairment of Mitochondrial Respiratory Chain Function and Insulin Signaling in Bovine Hepatocytes. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19061691"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"174959","DOI":"10.1016\/j.ejphar.2022.174959","article-title":"The lactate receptor GPR81 mediates hepatic lipid metabolism and the therapeutic effect of metformin on experimental NAFLDs","volume":"924","author":"Wu","year":"2022","journal-title":"Eur. J. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1002\/jcp.29026","article-title":"SCD1 regulates the AMPK\/SIRT1 pathway and histone acetylation through changes in adenine nucleotide metabolism in skeletal muscle","volume":"235","author":"Dziewulska","year":"2020","journal-title":"J. Cell. Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Zhang, B., Yang, W., Wang, S., Liu, R., Loor, J.J., Dong, Z., Zhao, Y., Ma, X., Xia, C., and Xu, C. (2020). Lipid Accumulation and Injury in Primary Calf Hepatocytes Challenged with Different Long-Chain Fatty Acids. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.547047"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Piccinin, E., Cariello, M., De Santis, S., Ducheix, S., Sabba, C., Ntambi, J.M., and Moschetta, A. (2019). Role of Oleic Acid in the Gut-Liver Axis: From Diet to the Regulation of Its Synthesis via Stearoyl-CoA Desaturase 1 (SCD1). Nutrients, 11.","DOI":"10.3390\/nu11102283"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2393","DOI":"10.1093\/emboj\/19.11.2393","article-title":"Signaling from Ras to Rac and beyond: Not just a matter of GEFs","volume":"19","author":"Scita","year":"2000","journal-title":"EMBO J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1700223","DOI":"10.1002\/bies.201700223","article-title":"Ubiquitylation Pathways In Insulin Signaling and Organismal Homeostasis","volume":"40","author":"Balaji","year":"2018","journal-title":"Bioessays"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.2147\/DMSO.S206867","article-title":"Liraglutide ameliorates nonalcoholic fatty liver disease in diabetic mice via the IRS2\/PI3K\/Akt signaling pathway","volume":"12","author":"Yang","year":"2019","journal-title":"Diabetes Metab. Syndr. Obes."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2193","DOI":"10.1007\/s11033-018-4380-8","article-title":"The role of DENND1A and CYP19A1 gene variants in individual susceptibility to obesity in Turkish population\u2014A preliminary study","volume":"45","author":"Kadioglu","year":"2018","journal-title":"Mol. Biol. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"111194","DOI":"10.1016\/j.celrep.2022.111194","article-title":"S-palmitoylation of PCSK9 induces sorafenib resistance in liver cancer by activating the PI3K\/AKT pathway","volume":"40","author":"Sun","year":"2022","journal-title":"Cell Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1016\/j.bbalip.2014.11.008","article-title":"Phosphoinositides: Key modulators of energy metabolism","volume":"1851","author":"Bridges","year":"2015","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Chang, J.G., Tien, N., Chang, Y.C., Lin, M.L., and Chen, S.S. (2021). Oxidative Stress-Induced Unscheduled CDK1-Cyclin B1 Activity Impairs ER-Mitochondria-Mediated Bioenergetic Metabolism. Cells, 10.","DOI":"10.3390\/cells10061280"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"8413032","DOI":"10.1155\/2016\/8413032","article-title":"The Tumorigenic Roles of the Cellular REDOX Regulatory Systems","volume":"2016","author":"Castaldo","year":"2016","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Castaldo, S.A., Ajime, T., Serrao, G., Anastacio, F., Rosa, J.T., Giacomantonio, C.A., Howarth, A., Hill, R., and Madureira, P.A. (2019). Annexin A2 Regulates AKT Upon H(2)O(2)-Dependent Signaling Activation in Cancer Cells. Cancers, 11.","DOI":"10.3390\/cancers11040492"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhang, B., Li, M., Zou, Y., Guo, H., Zhang, B., Xia, C., Zhang, H., Yang, W., and Xu, C. (2019). NFkappaB\/Orai1 Facilitates Endoplasmic Reticulum Stress by Oxidative Stress in the Pathogenesis of Non-alcoholic Fatty Liver Disease. Front. Cell Dev. Biol., 7.","DOI":"10.3389\/fcell.2019.00290"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.jsbmb.2014.10.014","article-title":"NEFAs activate the oxidative stress-mediated NF-kappaB signaling pathway to induce inflammatory response in calf hepatocytes","volume":"145","author":"Shi","year":"2015","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"153466","DOI":"10.1016\/j.phymed.2021.153466","article-title":"Arbutin attenuates LPS-induced acute kidney injury by inhibiting inflammation and apoptosis via the PI3K\/Akt\/Nrf2 pathway","volume":"82","author":"Zhang","year":"2021","journal-title":"Phytomedicine"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1038\/cr.2010.178","article-title":"Crosstalk of reactive oxygen species and NF-kappaB signaling","volume":"21","author":"Morgan","year":"2011","journal-title":"Cell Res."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Huang, Q.Y., Lai, X.N., Qian, X.L., Lv, L.C., Li, J., Duan, J., Xiao, X.H., and Xiong, L.X. (2019). Cdc42: A Novel Regulator of Insulin Secretion and Diabetes-Associated Diseases. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20010179"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1002\/hep.22610","article-title":"Hepatocyte-specific deletion of Cdc42 results in delayed liver regeneration after partial hepatectomy in mice","volume":"49","author":"Yuan","year":"2009","journal-title":"Hepatology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/j.jhep.2011.03.019","article-title":"Cdc42 and Rac1 are major contributors to the saturated fatty acid-stimulated JNK pathway in hepatocytes","volume":"56","author":"Sharma","year":"2012","journal-title":"J. Hepatol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1038\/sj.onc.1205036","article-title":"Rac1 and RhoG promote cell survival by the activation of PI3K and Akt, independently of their ability to stimulate JNK and NF-kappaB","volume":"21","author":"Murga","year":"2002","journal-title":"Oncogene"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1186\/s13567-019-0697-0","article-title":"Bovine ephemeral fever virus triggers autophagy enhancing virus replication via upregulation of the Src\/JNK\/AP1 and PI3K\/Akt\/NF-kappaB pathways and suppression of the PI3K\/Akt\/mTOR pathway","volume":"50","author":"Cheng","year":"2019","journal-title":"Vet. Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2259","DOI":"10.1253\/circj.CJ-16-0924","article-title":"Macrophage Apoptosis and Efferocytosis in the Pathogenesis of Atherosclerosis","volume":"80","author":"Linton","year":"2016","journal-title":"Circ. J."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e34051","DOI":"10.1016\/j.heliyon.2024.e34051","article-title":"Lactoferrin\u2019s role in modulating NF-\u03baB pathway to alleviate diabetes-associated inflammation: A novel in-silico study","volume":"10","author":"Arain","year":"2024","journal-title":"Heliyon"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"589","DOI":"10.18006\/2017.5(5).589.598","article-title":"Beneficial Impacts of Choline in Animal and Human with Special Reference to Its Role against Fatty Liver Syndrome","volume":"5","author":"Saeed","year":"2017","journal-title":"J. Exp. Biol. Agric. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1186\/s40104-021-00547-5","article-title":"Multifaceted role of one-carbon metabolism on immunometabolic control and growth during pregnancy, lactation and the neonatal period in dairy cattle","volume":"12","author":"Coleman","year":"2021","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"8305","DOI":"10.3168\/jds.2018-16204","article-title":"Hepatic betaine-homocysteine methyltransferase and methionine synthase activity and intermediates of the methionine cycle are altered by choline supply during negative energy balance in Holstein cows","volume":"102","author":"Coleman","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Chandler, T.L., and White, H.M. (2019). Glucose metabolism is differentially altered by choline and methionine in bovine neonatal hepatocytes. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0217160"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"120462","DOI":"10.1016\/j.atherosclerosis.2025.120462","article-title":"Effect of olezarsen on lipoprotein-associated ApoC-III levels in patients with familial chylomicronemia syndrome","volume":"408","author":"Yang","year":"2025","journal-title":"Atherosclerosis"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"G726","DOI":"10.1152\/ajpgi.00436.2015","article-title":"Liver-specific loss of Perilipin 2 alleviates diet-induced hepatic steatosis, inflammation, and fibrosis","volume":"310","author":"Najt","year":"2016","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1007\/s13238-018-0593-9","article-title":"Phosphorylation of PLIN3 by AMPK promotes dispersion of lipid droplets during starvation","volume":"10","author":"Zhu","year":"2019","journal-title":"Protein Cell"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1008","DOI":"10.1016\/j.bbrc.2017.11.134","article-title":"Fatty acid oxidation alleviates the energy deficiency caused by the loss of MPC1 in MPC1(+\/\u2212) mice","volume":"495","author":"Zou","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.cbpb.2015.08.010","article-title":"Molecular characterization of carnitine palmitoyltransferase IA in Megalobrama amblycephala and effects on its expression of feeding status and dietary lipid and berberine","volume":"191","author":"Lu","year":"2016","journal-title":"Comp. Biochem. Physiol. B Biochem. Mol. Biol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"8319","DOI":"10.3168\/jds.2019-16406","article-title":"Choline supply during negative nutrient balance alters hepatic cystathionine beta-synthase, intermediates of the methionine cycle and transsulfuration pathway, and liver function in Holstein cows","volume":"102","author":"Coleman","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/1476-511X-12-34","article-title":"Betaine supplement alleviates hepatic triglyceride accumulation of apolipoprotein E deficient mice via reducing methylation of peroxisomal proliferator-activated receptor alpha promoter","volume":"12","author":"Wang","year":"2013","journal-title":"Lipids Health Dis."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"3238","DOI":"10.3390\/cimb44070223","article-title":"How Different Dietary Methionine Sources Could Modulate the Hepatic Metabolism in Rainbow Trout?","volume":"44","author":"Ceccotti","year":"2022","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"2552","DOI":"10.3390\/nu6072552","article-title":"The effects of choline on hepatic lipid metabolism, mitochondrial function and antioxidative status in human hepatic C3A cells exposed to excessive energy substrates","volume":"6","author":"Zhu","year":"2014","journal-title":"Nutrients"},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Chandler, T.L., and White, H.M. (2017). Choline and methionine differentially alter methyl carbon metabolism in bovine neonatal hepatocytes. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0171080"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"2458","DOI":"10.1194\/jlr.M046961","article-title":"Acyl-CoA thioesterase-2 facilitates mitochondrial fatty acid oxidation in the liver","volume":"55","author":"Moffat","year":"2014","journal-title":"J. Lipid Res."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1093\/oxfordjournals.jbchem.a022240","article-title":"Fatty acid metabolism and very low density lipoprotein secretion in liver slices from rats and preruminant calves","volume":"124","author":"Graulet","year":"1998","journal-title":"J. Biochem."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"8844","DOI":"10.3168\/jds.2025-26349","article-title":"Hepatocyte necroptosis is associated with liver damage in dairy cows with ketosis","volume":"108","author":"Liu","year":"2025","journal-title":"J. Dairy Sci."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/16\/7\/451\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T09:56:52Z","timestamp":1782727012000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/16\/7\/451"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,26]]},"references-count":70,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["metabo16070451"],"URL":"https:\/\/doi.org\/10.3390\/metabo16070451","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,26]]}}}