Hepatotoxicity of Nonesterified Fatty Acids to Dairy Cows: Pathophysiological Mechanisms and Prospective Solutions

Abstract

Circulating concentrations of nonesterified fatty acids (NEFAs) are elevated due to lipid mobilization from adipose tissue in periparturient dairy cows. Although this metabolic adaptation facilitates energy homeostasis under the negative energy balance condition, sustained systemic NEFA overload induces profound hepatic impairment. Emerging evidence identifies excessive NEFAs to be the pathophysiological cornerstone of periparturient disorders; however, the precise molecular mechanisms underlying NEFA-induced hepatotoxicity remain incompletely characterized, hindering the development of effective preventive and therapeutic strategies. This literature review synthesizes contemporary insights into key cellular pathways implicated in NEFA-mediated hepatotoxicity: disorders in lipid and carbohydrate metabolism, impairment of autophagy, excessive inflammatory response, mitochondrial dysfunction, oxidative stress, endoplasmic reticulum stress, and finally, cell death. Critical analysis reveals two underexplored dimensions in current research paradigms: (1) The dynamic composition of circulating NEFAs modulates hepatotoxic potency through differential membrane incorporation and signaling pathway activation, suggesting that improving blood NEFA composition through dietary fat supplementation offers a potential strategy; and (2) the periparturient inflammatory milieu potentiates NEFA toxicity, suggesting targeted anti-inflammatory interventions ameliorate transition period adaptation. Consequently, this review advances our mechanistic understanding while providing translational frameworks for improving periparturient management through precision nutrition and therapeutic development.

Keywords

autophagy / cell death / endoplasmic reticulum stress / inflammation / lipid and carbohydrate metabolism disorder / mitochondrial dysfunction / oxidative stress

References

[1] F. Ceciliani, C. Lecchi, C. Urh, and H. Sauerwein, “Proteomics and Metabolomics Characterizing the Pathophysiology of Adaptive Reactions to the Metabolic Challenges During the Transition From Late Pregnancy to Early Lactation in Dairy Cows,” Journal of Proteomics 178 (2018): 92–106, https://doi.org/10.1016/j.jprot.2017.10.010.

[2] R. Mikuła, E. Pruszyńska-Oszmałek, M. Pszczola, et al., “Changes in Metabolic and Hormonal Profiles During Transition Period in Dairy Cattle—The Role of Spexin,” BMC Veterinary Research 17, no. 1 (2021): 359, https://doi.org/10.1186/s12917-021-03069-4.

[3] N. Alsabeeh, B. Chausse, P. A. Kakimoto, A. J. Kowaltowski, and O. Shirihai, “Cell Culture Models of Fatty Acid Overload: Problems and Solutions,” Biochimica et Biophysica Acta, Molecular and Cell Biology of Lipids 1863, no. 2 (2018): 143–151, https://doi.org/10.1016/j.bbalip.2017.11.006.

[4] Z. Cheng, A. Wylie, C. Ferris, K. Ingvartsen, and D. Wathes, “Effect of Diet and Nonesterified Fatty Acid Levels on Global Transcriptomic Profiles in Circulating Peripheral Blood Mononuclear Cells in Early Lactation Dairy Cows,” Journal of Dairy Science 104, no. 9 (2021): 10059–10075, https://doi.org/10.3168/jds.2021-20136.

[5] G. C. Farrell, F. Haczeyni, and S. Chitturi, “Pathogenesis of NASH: How Metabolic Complications of Overnutrition Favour Lipotoxicity and Pro-Inflammatory Fatty Liver Disease,” Advances in Experimental Medicine and Biology 1061 (2018): 19–44, https://doi.org/10.1007/978-981-10-8684-7_3.

[6] C. Zhao, J. Li, M. Liu, et al., “Inhibition of Cluster Antigen 36 Protects Against Fatty Acid-Induced Lipid Accumulation, Oxidative Stress, and Inflammation in Bovine Hepatocytes,” Journal of Dairy Science 106, no. 12 (2023): 9186–9199, https://doi.org/10.3168/jds.2023-23282.

[7] J. A. McArt, D. V. Nydam, G. R. Oetzel, T. R. Overton, and P. A. Ospina, “Elevated Non-Esterified Fatty Acids and Β-Hydroxybutyrate and Their Association With Transition Dairy Cow Performance,” Veterinary Journal 198, no. 3 (2013): 560–570, https://doi.org/10.1016/j.tvjl.2013.08.011.

[8] I. Nicola, H. Chupin, J. P. Roy, et al., “Association Between Prepartum Nonesterified Fatty Acid Serum Concentrations and Postpartum Diseases in Dairy Cows,” Journal of Dairy Science 105, no. 11 (2022): 9098–9106, https://doi.org/10.3168/jds.2022-22014.

[9] N. Chapinal, M. E. Carson, S. J. LeBlanc, et al., “The Association of Serum Metabolites in the Transition Period With Milk Production and Early-Lactation Reproductive Performance,” Journal of Dairy Science 95, no. 3 (2012): 1301–1309, https://doi.org/10.3168/jds.2011-4724.

[10] S. J. LeBlanc, K. E. Leslie, and T. F. Duffield, “Metabolic Predictors of Displaced Abomasum in Dairy Cattle,” Journal of Dairy Science 88, no. 1 (2005): 159–170, https://doi.org/10.3168/jds.S0022-0302(05)72674-6.

[11] J. A. McArt, D. V. Nydam, and G. R. Oetzel, “Dry Period and Parturient Predictors of Early Lactation Hyperketonemia in Dairy Cattle,” Journal of Dairy Science 96, no. 1 (2013): 198–209, https://doi.org/10.3168/jds.2012-5681.

[12] P. A. Ospina, D. V. Nydam, T. Stokol, and T. Overton, “Association Between the Proportion of Sampled Transition Cows With Increased Nonesterified Fatty Acids and Beta-Hydroxybutyrate and Disease Incidence, Pregnancy Rate, and Milk Production at the Herd Level,” Journal of Dairy Science 93, no. 8 (2010): 3595–3601, https://doi.org/10.3168/jds.2010-3074.

[13] H. A. Seifi, S. J. Leblanc, K. E. Leslie, and T. F. Duffield, “Metabolic Predictors of Post-Partum Disease and Culling Risk in Dairy Cattle,” Veterinary Journal 188, no. 2 (2011): 216–220, https://doi.org/10.1016/j.tvjl.2010.04.007.

[14] E. S. Ribeiro, F. S. Lima, L. F. Greco, et al., “Prevalence of Periparturient Diseases and Effects on Fertility of Seasonally Calving Grazing Dairy Cows Supplemented With Concentrates,” Journal of Dairy Science 96, no. 9 (2013): 5682–5697, https://doi.org/10.3168/jds.2012-6335.

[15] D. Liang, L. M. Arnold, C. J. Stowe, R. Harmon, and J. Bewley, “Estimating Us Dairy Clinical Disease Costs With a Stochastic Simulation Model,” Journal of Dairy Science 100, no. 2 (2017): 1472–1486, https://doi.org/10.3168/jds.2016-11565.

[16] D. Raboisson, M. Mounié, and E. Maigné, “Diseases, Reproductive Performance, and Changes in Milk Production Associated With Subclinical Ketosis in Dairy Cows: A Meta-Analysis and Review,” Journal of Dairy Science 97, no. 12 (2014): 7547–7563, https://doi.org/10.3168/jds.2014-8237.

[17] P. A. Ospina, D. V. Nydam, T. Stokol, and T. Overton, “Evaluation of Nonesterified Fatty Acids and Beta-Hydroxybutyrate in Transition Dairy Cattle in the Northeastern United States: Critical Thresholds for Prediction of Clinical Diseases,” Journal of Dairy Science 93, no. 2 (2010): 546–554, https://doi.org/10.3168/jds.2009-2277.

[18] N. Chapinal, S. J. Leblanc, M. E. Carson, et al., “Herd-Level Association of Serum Metabolites in the Transition Period With Disease, Milk Production, and Early Lactation Reproductive Performance,” Journal of Dairy Science 95, no. 10 (2012): 5676–5682, https://doi.org/10.3168/jds.2011-5132.

[19] T. Vanholder, J. Papen, R. Bemers, G. Vertenten, and A. Berge, “Risk Factors for Subclinical and Clinical Ketosis and Association With Production Parameters in Dairy Cows in the Netherlands,” Journal of Dairy Science 98, no. 2 (2015): 880–888, https://doi.org/10.3168/jds.2014-8362.

[20] G. A. Contreras, N. J. O’Boyle, T. H. Herdt, and L. Sordillo, “Lipomobilization in Periparturient Dairy Cows Influences the Composition of Plasma Nonesterified Fatty Acids and Leukocyte Phospholipid Fatty Acids,” Journal of Dairy Science 93, no. 6 (2010): 2508–2516, https://doi.org/10.3168/jds.2009-2876.

[21] D. S. Hammon, I. M. Evjen, T. R. Dhiman, J. Goff, and J. Walters, “Neutrophil Function and Energy Status in Holstein Cows With Uterine Health Disorders,” Veterinary Immunology and Immunopathology 113, no. 1–2 (2006): 21–29, https://doi.org/10.1016/j.vetimm.2006.03.022.

[22] A. Benedet, C. L. Manuelian, A. Zidi, M. Penasa, and M. De Marchi, “Invited Review: Β-Hydroxybutyrate Concentration in Blood and Milk and Its Associations With Cow Performance,” Animal 13, no. 8 (2019): 1676–1689, https://doi.org/10.1017/S175173111900034X.

[23] X. Li, Y. Li, W. Yang, et al., “SREBP-1c Overexpression Induces Triglycerides Accumulation Through Increasing Lipid Synthesis and Decreasing Lipid Oxidation and VLDL Assembly in Bovine Hepatocytes,” Journal of Steroid Biochemistry and Molecular Biology 143 (2014): 174–182, https://doi.org/10.1016/j.jsbmb.2014.02.009.

[24] C. Zhao, B. Wu, J. Li, et al., “Adiporon Alleviates Fatty Acid-Induced Lipid Accumulation and Mitochondrial Dysfunction in Bovine Hepatocytes by Promoting Autophagy,” Journal of Dairy Science 106, no. 8 (2023): 5763–5774, https://doi.org/10.3168/jds.2022-22723.

[25] L. Liu, X. Li, Y. Li, et al., “Effects of Nonesterified Fatty Acids on the Synthesis and Assembly of Very Low Density Lipoprotein in Bovine Hepatocytes In Vitro,” Journal of Dairy Science 97, no. 3 (2014): 1328–1335, https://doi.org/10.3168/jds.2013-6654.

[26] H. Jia, X. Li, G. Liu, et al., “Perilipin 5 Promotes Hepatic Steatosis in Dairy Cows Through Increasing Lipid Synthesis and Decreasing Very Low Density Lipoprotein Assembly,” Journal of Dairy Science 102, no. 1 (2019): 833–845, https://doi.org/10.3168/jds.2018-15208.

[27] W. Yang, S. Wang, J. J. Loor, et al., “Role of Sortilin 1 (SORT1) on Lipid Metabolism in Bovine Liver,” Journal of Dairy Science 105, no. 6 (2021): 5420–5434, https://doi.org/10.3168/jds.2021-21607.

[28] L. Liu, D. Xing, X. Du, et al., “Sirtuin 3 Improves Fatty Acid Metabolism in Response to High Nonesterified Fatty Acids in Calf Hepatocytes by Modulating Gene Expression,” Journal of Dairy Science 103, no. 7 (2020): 6557–6568, https://doi.org/10.3168/jds.2019-17670.

[29] D. Xing, B. Wang, H. Lu, et al., “Sirtuin 3 Restores Synthesis and Secretion of Very Low-Density Lipoproteins in Cow Hepatocytes Challenged With Nonesterified Fatty Acids In Vitro,” Veterinary Sciences 8, no. 7 (2021): 121, https://doi.org/10.3390/vetsci8070121.

[30] B. Zhao, C. Luo, M. Zhang, et al., “Knockdown of Phosphatase and Tensin Homolog (PTEN) Inhibits Fatty Acid Oxidation and Reduces Very Low Density Lipoprotein Assembly and Secretion in Calf Hepatocytes,” Journal of Dairy Science 103, no. 11 (2020): 10728–10741, https://doi.org/10.3168/jds.2019-17920.

[31] J. Dong, J. J. Loor, R. Zuo, et al., “Low Abundance of Mitofusin 2 in Dairy Cows With Moderate Fatty Liver Is Associated With Alterations in Hepatic Lipid Metabolism,” Journal of Dairy Science 102, no. 8 (2019): 7536–7547, https://doi.org/10.3168/jds.2019-16544.

[32] R. R. Grummer, “Etiology of Lipid-Related Metabolic Disorders in Periparturient Dairy Cows,” Journal of Dairy Science 76, no. 12 (1993): 3882–3896, https://doi.org/10.3168/jds.S0022-0302(93)77729-2.

[33] A. K. Leamy, R. A. Egnatchik, and J. D. Young, “Molecular Mechanisms and the Role of Saturated Fatty Acids in the Progression of Non-Alcoholic Fatty Liver Disease,” Progress in Lipid Research 52, no. 1 (2013): 165–174, https://doi.org/10.1016/j.plipres.2012.10.004.

[34] J. R. Aschenbach, N. B. Kristensen, S. S. Donkin, H. M. Hammon, and G. B. Penner, “Gluconeogenesis in Dairy Cows: The Secret of Making Sweet Milk From Sour Dough,” IUBMB Life 62, no. 12 (2010): 869–877, https://doi.org/10.1002/iub.400.

[35] Q. Zhang, S. L. Koser, B. J. Bequette, and S. S. Donkin, “Effect of Propionate on mRNA Expression of Key Genes for Gluconeogenesis in Liver of Dairy Cattle,” Journal of Dairy Science 98, no. 12 (2015): 8698–8709, https://doi.org/10.3168/jds.2015-9590.

[36] C. K. Reynolds, P. C. Aikman, B. Lupoli, D. Humphries, and D. Beever, “Splanchnic Metabolism of Dairy Cows During the Transition From Late Gestation Through Early Lactation,” Journal of Dairy Science 86, no. 4 (2003): 1201–1217, https://doi.org/10.3168/jds.S0022-0302(03)73704-7.

[37] A. Murondoti, R. Jorritsma, A. C. Beynen, T. Wensing, and M. J. Geelen, “Activities of the Enzymes of Hepatic Gluconeogenesis in Periparturient Dairy Cows With Induced Fatty Liver,” Journal of Dairy Research 71, no. 2 (2004): 129–134, https://doi.org/10.1017/s0022029904000020.

[38] X. Li, X. Li, G. Bai, et al., “Effects of Non-Esterified Fatty Acids on the Gluconeogenesis in Bovine Hepatocytes,” Molecular and Cellular Biochemistry 359, no. 1–2 (2012): 385–388, https://doi.org/10.1007/s11010-011-1032-x.

[39] T. Rukkwamsuk, T. Wensing, and M. J. Geelen, “Effect of Fatty Liver on Hepatic Gluconeogenesis in Periparturient Dairy Cows,” Journal of Dairy Science 82, no. 3 (1999): 500–505, https://doi.org/10.3168/jds.S0022-0302(99)75260-4.

[40] D. Glick, S. Barth, and K. F. Macleod, “Autophagy: Cellular and Molecular Mechanisms,” Journal of Pathology 221, no. 1 (2010): 3–12, https://doi.org/10.1002/path.2697.

[41] P. E. Rautou, A. Mansouri, D. Lebrec, F. Durand, D. Valla, and R. Moreau, “Autophagy in Liver Diseases,” Journal of Hepatology 53, no. 6 (2010): 1123–1134, https://doi.org/10.1016/j.jhep.2010.07.006.

[42] W. Li, P. He, Y. Huang, et al., “Selective Autophagy of Intracellular Organelles: Recent Research Advances,” Theranostics 11, no. 1 (2021): 222–256, https://doi.org/10.7150/thno.49860.

[43] M. Chen, J. J. Loor, Q. Zhai, et al., “Short Communication: Enhanced Autophagy Activity in Liver Tissue of Dairy Cows With Mild Fatty Liver,” Journal of Dairy Science 103, no. 4 (2020): 3628–3635, https://doi.org/10.3168/jds.2019-17457.

[44] X. Du, G. Liu, J. J. Loor, et al., “Impaired Hepatic Autophagic Activity in Dairy Cows With Severe Fatty Liver Is Associated With Inflammation and Reduced Liver Function,” Journal of Dairy Science 101, no. 12 (2018): 11175–11185, https://doi.org/10.3168/jds.2018-15120.

[45] X. Du, M. Chen, Z. Fang, et al., “Evaluation of Hepatic AMPK, Mtorc1, and Autophagy-Lysosomal Pathway in Cows With Mild or Moderate Fatty Liver,” Journal of Dairy Science 107, no. 5 (2024): 3269–3279, https://doi.org/10.3168/jds.2023-24000.

[46] T. Shen, X. Li, B. Jin, et al., “Free Fatty Acids Impair Autophagic Activity and Activate Nuclear Factor Kappa B Signaling and NLR Family Pyrin Domain Containing 3 Inflammasome in Calf Hepatocytes,” Journal of Dairy Science 104, no. 11 (2021): 11973–11982, https://doi.org/10.3168/jds.2021-20273.

[47] Z. Fang, G. Liu, M. Zhu, et al., “Low Abundance of Mitophagy Markers Is Associated With Reactive Oxygen Species Overproduction in Cows With Fatty Liver and Causes Reactive Oxygen Species Overproduction and Lipid Accumulation in Calf Hepatocytes,” Journal of Dairy Science 105, no. 9 (2022): 7829–7841, https://doi.org/10.3168/jds.2021-21774.

[48] B. J. Bradford and T. H. Swartz, “Review: Following the Smoke Signals: Inflammatory Signaling in Metabolic Homeostasis and Homeorhesis in Dairy Cattle,” supplement, Animal 14, no. S1 (2020): s144–s154, https://doi.org/10.1017/S1751731119003203.

[49] B. J. Bradford, K. Yuan, J. K. Farney, L. Mamedova, and A. Carpenter, “Invited Review: Inflammation During the Transition to Lactation: New Adventures With an Old Flame,” Journal of Dairy Science 98, no. 10 (2015): 6631–6650, https://doi.org/10.3168/jds.2015-9683.

[50] X. Shi, D. Li, Q. Deng, et al., “NEFAs Activate the Oxidative Stress-Mediated NF-κB Signaling Pathway to Induce Inflammatory Response in Calf Hepatocytes,” Journal of Steroid Biochemistry and Molecular Biology 145 (2015): 103–112, https://doi.org/10.1016/j.jsbmb.2014.10.014.

[51] X. Li, W. Huang, J. Gu, et al., “SREBP-1c Overactivates Ros-Mediated Hepatic NF-κB Inflammatory Pathway in Dairy Cows With Fatty Liver,” Cellular Signalling 27, no. 10 (2015): 2099–2109, https://doi.org/10.1016/j.cellsig.2015.07.011.

[52] J. Dong, G. Bobe, Y. Guan, et al., “Mitochondrial Membrane Protein Mitofusin 2 as a Potential Therapeutic Target for Treating Free Fatty Acid-Induced Hepatic Inflammation in Dairy Cows During Early Lactation,” Journal of Dairy Science 103, no. 6 (2020): 5561–5574, https://doi.org/10.3168/jds.2019-17652.

[53] X. Li, W. Zhang, Q. Cao, et al., “Mitochondrial Dysfunction in Fibrotic Diseases,” Cell Death Discovery 6, no. 1 (2020): 80, https://doi.org/10.1038/s41420-020-00316-9.

[54] S. Miwa, S. Kashyap, E. Chini, and T. von Zglinicki, “Mitochondrial Dysfunction in Cell Senescence and Aging,” Journal of Clinical Investigation 132, no. 13 (2022): e158447, https://doi.org/10.1172/JCI158447.

[55] W. Gao, X. Du, L. Lei, et al., “NEFA-Induced ROS Impaired Insulin Signalling Through the JNK and p38MAPK Pathways in Non-Alcoholic Steatohepatitis,” Journal of Cellular and Molecular Medicine 22, no. 7 (2018): 3408–3422, https://doi.org/10.1111/jcmm.13617.

[56] X. Wang, M. Zhu, J. J. Loor, et al., “Propionate Alleviates Fatty Acid-Induced Mitochondrial Dysfunction, Oxidative Stress, and Apoptosis by Upregulating PPARG Coactivator 1 Alpha in Hepatocytes,” Journal of Dairy Science 105, no. 5 (2022): 4581–4592, https://doi.org/10.3168/jds.2021-21198.

[57] B. Zhang, M. Li, W. Yang, et al., “Mitochondrial Dysfunction and Endoplasmic Reticulum Stress in Calf Hepatocytes Are Associated With Fatty Acid-Induced ORAI Calcium Release-Activated Calcium Modulator 1 Signaling,” Journal of Dairy Science 103, no. 12 (2020): 11945–11956, https://doi.org/10.3168/jds.2020-18684.

[58] M. Cnop, F. Foufelle, and L. A. Velloso, “Endoplasmic Reticulum Stress, Obesity and Diabetes,” Trends in Molecular Medicine 18, no. 1 (2012): 59–68, https://doi.org/10.1016/j.molmed.2011.07.010.

[59] D. K. Gessner, G. Schlegel, R. Ringseis, F. J. Schwarz, and K. Eder, “Up-Regulation of Endoplasmic Reticulum Stress Induced Genes of the Unfolded Protein Response in the Liver of Periparturient Dairy Cows,” BMC Veterinary Research 10, no. 1 (2014): 46, https://doi.org/10.1186/1746-6148-10-46.

[60] Y. Huang, C. Zhao, Y. Kong, et al., “Elucidation of the Mechanism of NEFA-Induced PERK-eIF2α Signaling Pathway Regulation of Lipid Metabolism in Bovine Hepatocytes,” Journal of Steroid Biochemistry and Molecular Biology 211 (2021): 105893, https://doi.org/10.1016/j.jsbmb.2021.105893.

[61] Z. Fang, W. Gao, Q. Jiang, et al., “Targeting IRE1α and PERK in the Endoplasmic Reticulum Stress Pathway Attenuates Fatty Acid-Induced Insulin Resistance in Bovine Hepatocytes,” Journal of Dairy Science 105, no. 8 (2022): 6895–6908, https://doi.org/10.3168/jds.2021-21754.

[62] Y. Huang, C. Zhao, Y. Liu, et al., “NEFA Promotes Autophagosome Formation Through Modulating PERK Signaling Pathway in Bovine Hepatocytes,” Animals (Basel) 11, no. 12 (2021): 3400, https://doi.org/10.3390/ani11123400.

[63] Y. Zhu, Y. Guan, J. J. Loor, et al., “Fatty Acid-Induced Endoplasmic Reticulum Stress Promoted Lipid Accumulation in Calf Hepatocytes, and Endoplasmic Reticulum Stress Existed in the Liver of Severe Fatty Liver Cows,” Journal of Dairy Science 102, no. 8 (2019): 7359–7370, https://doi.org/10.3168/jds.2018-16015.

[64] M. Li, W. Yang, J. Wen, et al., “Intracellular Ca2⁺ Signaling and ORAI Calcium Release-Activated Calcium Modulator 1 Are Associated With Hepatic Lipidosis in Dairy Cattle,” Journal of Animal Science 99, no. 7 (2021): skab184, https://doi.org/10.1093/jas/skab184.

[65] M. Li, B. Zhao, J. Wang, et al., “Caveolin 1 in Bovine Liver Is Associated With Fatty Acid-Induced Lipid Accumulation and the Endoplasmic Reticulum Unfolded Protein Response: Role in Fatty Liver Development,” Journal of Dairy Science 108, no. 1 (2025): 1007–1021, https://doi.org/10.3168/jds.2024-25349.

[66] M. A. Islam, S. Adachi, Y. Shiiba, K. i. Takeda, S. Haga, and S. Yonekura, “Effects of Starvation-Induced Negative Energy Balance on Endoplasmic Reticulum Stress in the Liver of Cows,” Animal Bioscience 35, no. 1 (2022): 22–28, https://doi.org/10.5713/ab.21.0140.

[67] R. Ringseis, D. K. Gessner, and K. Eder, “Molecular Insights into the Mechanisms of Liver-Associated Diseases in Early-Lactating Dairy Cows: Hypothetical Role of Endoplasmic Reticulum Stress,” Journal of Animal Physiology and Animal Nutrition 99, no. 4 (2015): 626–645, https://doi.org/10.1111/jpn.12263.

[68] J. L. Martindale and N. J. Holbrook, “Cellular Response to Oxidative Stress: Signaling for Suicide and Survival,” Journal of Cellular Physiology 192, no. 1 (2002): 1–15, https://doi.org/10.1002/jcp.10119.

[69] H. J. Forman and H. Zhang, “Targeting Oxidative Stress in Disease: Promise and Limitations of Antioxidant Therapy,” Nature Reviews Drug Discovery 20, no. 9 (2021): 689–709, https://doi.org/10.1038/s41573-021-00233-1.

[70] P. Poprac, K. Jomova, M. Simunkova, V. Kollar, C. J. Rhodes, and M. Valko, “Targeting Free Radicals in Oxidative Stress-Related Human Diseases,” Trends in Pharmacological Sciences 38, no. 7 (2017): 592–607, https://doi.org/10.1016/j.tips.2017.04.005.

[71] H. Sejersen, M. T. Sørensen, T. Larsen, E. Bendixen, and K. Ingvartsen, “Liver Protein Expression in Dairy Cows With High Liver Triglycerides in Early Lactation,” Journal of Dairy Science 95, no. 5 (2012): 2409–2421, https://doi.org/10.3168/jds.2011-4604.

[72] A. Abuelo, J. Hernández, J. L. Benedito, and C. Castillo, “The Importance of the Oxidative Status of Dairy Cattle in the Periparturient Period: Revisiting Antioxidant Supplementation,” Journal of Animal Physiology and Animal Nutrition 99, no. 6 (2015): 1003–1016, https://doi.org/10.1111/jpn.12273.

[73] U. Bernabucci, B. Ronchi, N. Lacetera, and A. Nardone, “Influence of Body Condition Score on Relationships Between Metabolic Status and Oxidative Stress in Periparturient Dairy Cows,” Journal of Dairy Science 88, no. 6 (2005): 2017–2026, https://doi.org/10.3168/jds.S0022-0302(05)72878-2.

[74] C. Zhang, Q. Shao, M. Liu, et al., “Liver Fibrosis Is a Common Pathological Change in the Liver of Dairy Cows With Fatty Liver,” Journal of Dairy Science 106, no. 4 (2023): 2700–2715, https://doi.org/10.3168/jds.2022-22021.

[75] Y. Li, H. Y. Ding, X. C. Wang, et al., “An Association Between the Level of Oxidative Stress and the Concentrations of NEFA and BHBA in the Plasma of Ketotic Dairy Cows,” Journal of Animal Physiology and Animal Nutrition 100, no. 5 (2016): 844–851, https://doi.org/10.1111/jpn.12454.

[76] Y. Song, X. Li, Y. Li, et al., “Non-Esterified Fatty Acids Activate the ROS-p38-p53/Nrf2 Signaling Pathway to Induce Bovine Hepatocyte Apoptosis In Vitro,” Apoptosis 19, no. 6 (2014): 984–997, https://doi.org/10.1007/s10495-014-0982-3.

[77] Z. Fang, X. Jiang, S. Wang, et al., “Nuciferine Protects Bovine Hepatocytes Against Free Fatty Acid-Induced Oxidative Damage by Activating the Transcription Factor EB/Peroxisome Proliferator-Activated Receptor γ Coactivator 1 Alpha Pathway,” Journal of Dairy Science 107, no. 1 (2024): 625–640, https://doi.org/10.3168/jds.2022-22801.

[78] M. J. Kuhn, V. Mavangira, and L. M. Sordillo, “Invited Review: Cytochrome P450 Enzyme Involvement in Health and Inflammatory-Based Diseases of Dairy Cattle,” Journal of Dairy Science 104, no. 2 (2021): 1276–1290, https://doi.org/10.3168/jds.2020-18997.

[79] Z. Peng, X. Li, Z. Wang, and G. Liu, “The Effects of Non-Esterified Fatty Acids and Β-Hydroxybutyrate on the Hepatic CYP2E1 in Cows With Clinical Ketosis,” Journal of Dairy Research 86, no. 1 (2019): 68–72, https://doi.org/10.1017/S0022029919000025.

[80] S. Ghavami, S. Shojaei, B. Yeganeh, et al., “Autophagy and Apoptosis Dysfunction in Neurodegenerative Disorders,” Progress in Neurobiology 112 (2014): 24–49, https://doi.org/10.1016/j.pneurobio.2013.10.004.

[81] L. Galluzzi, I. Vitale, S. A. Aaronson, et al., “Molecular Mechanisms of Cell Death: Recommendations of the Nomenclature Committee on Cell Death 2018,” Cell Death & Differentiation 25, no. 3 (2018): 486–541, https://doi.org/10.1038/s41418-017-0012-4.

[82] S. Shen, Y. Shao, and C. Li, “Different Types of Cell Death and Their Shift in Shaping Disease,” Cell Death Discovery 9, no. 1 (2023): 284, https://doi.org/10.1038/s41420-023-01581-0.

[83] N. Ketelut-Carneiro and K. A. Fitzgerald, “Apoptosis, Pyroptosis, and Necroptosis-Oh My! The Many Ways a Cell Can Die,” Journal of Molecular Biology 434, no. 4 (2022): 167378, https://doi.org/10.1016/j.jmb.2021.167378.

[84] M. Tharwat, A. Takamizawa, Y. Z. Hosaka, D. Endoh, and S. Oikawa, “Hepatocyte Apoptosis in Dairy Cattle During the Transition Period,” Canadian Journal of Veterinary Research 76, no. 4 (2012): 241–247.

[85] M. Tharwat, D. Endoh, and S. Oikawa, “Hepatocyte Apoptosis in Dairy Cows With Fatty Infiltration of the Liver,” Research in Veterinary Science 93, no. 3 (2012): 1281–1286, https://doi.org/10.1016/j.rvsc.2012.03.011.

[86] X. Du, L. Chen, D. Huang, et al., “Elevated Apoptosis in the Liver of Dairy Cows With Ketosis,” Cellular Physiology and Biochemistry 43, no. 2 (2017): 568–578, https://doi.org/10.1159/000480529.

[87] Z. Rao, Y. Zhu, P. Yang, et al., “Pyroptosis in Inflammatory Diseases and Cancer,” Theranostics 12, no. 9 (2022): 4310–4329, https://doi.org/10.7150/thno.71086.

[88] P. Broz, “Immunology: Caspase Target Drives Pyroptosis,” Nature 526, no. 7575 (2015): 642–643, https://doi.org/10.1038/nature15632.

[89] Y. Huang, W. Xu, and R. Zhou, “NLRP3 Inflammasome Activation and Cell Death,” Cellular and Molecular Immunology 18, no. 9 (2021): 2114–2127, https://doi.org/10.1038/s41423-021-00740-6.

[90] B. E. Burdette, A. N. Esparza, H. Zhu, and S. Wang, “Gasdermin D in Pyroptosis,” Acta Pharmaceutica Sinica B 11, no. 9 (2021): 2768–2782, https://doi.org/10.1016/j.apsb.2021.02.006.

[91] D. Frank and J. E. Vince, “Pyroptosis Versus Necroptosis: Similarities, Differences, and Crosstalk,” Cell Death & Differentiation 26, no. 1 (2019): 99–114, https://doi.org/10.1038/s41418-018-0212-6.

[92] A. Wree, A. Eguchi, M. D. McGeough, et al., “NLRP3 Inflammasome Activation Results in Hepatocyte Pyroptosis, Liver Inflammation, and Fibrosis in Mice,” Hepatology 59, no. 3 (2014): 898–910, https://doi.org/10.1002/hep.26592.

[93] S. Gaul, A. Leszczynska, F. Alegre, et al., “Hepatocyte Pyroptosis and Release of Inflammasome Particles Induce Stellate Cell Activation and Liver Fibrosis,” Journal of Hepatology 74, no. 1 (2021): 156–167, https://doi.org/10.1016/j.jhep.2020.07.041.

[94] J. Shi, W. Gao, and F. Shao, “Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death,” Trends in Biochemical Sciences 42, no. 4 (2017): 245–254, https://doi.org/10.1016/j.tibs.2016.10.004.

[95] T. Qiu, P. Pei, X. Yao, et al., “Taurine Attenuates Arsenic-Induced Pyroptosis and Nonalcoholic Steatohepatitis by Inhibiting the Autophagic-Inflammasomal Pathway,” Cell Death & Disease 9, no. 10 (2018): 946, https://doi.org/10.1038/s41419-018-1004-0.

[96] E. H. Koh, J. E. Yoon, M. S. Ko, et al., “Sphingomyelin Synthase 1 Mediates Hepatocyte Pyroptosis to Trigger Non-Alcoholic Steatohepatitis,” Gut 70, no. 10 (2021): 1954–1964, https://doi.org/10.1136/gutjnl-2020-322509.

[97] J. Knorr, A. Wree, and A. E. Feldstein, “Pyroptosis in Steatohepatitis and Liver Diseases,” Journal of Molecular Biology 434, no. 4 (2022): 167271, https://doi.org/10.1016/j.jmb.2021.167271.

[98] H. Zhang, H. Shi, W. Xie, et al., “Subacute Ruminal Acidosis Induces Pyroptosis via the Mitophagy-Mediated NLRP3 Inflammasome Activation in the Livers of Dairy Cows Fed a High-Grain Diet,” Journal of Dairy Science 107, no. 6 (2024): 4092–4107, https://doi.org/10.3168/jds.2023-23718.

[99] T. Shen, X. Li, J. J. Loor, et al., “Hepatic Nuclear Factor Kappa B Signaling Pathway and NLR Family Pyrin Domain Containing 3 Inflammasome Is Over-Activated in Ketotic Dairy Cows,” Journal of Dairy Science 102, no. 11 (2019): 10554–10563, https://doi.org/10.3168/jds.2019-16706.

[100] X. Chen, L. Li, X. Liu, et al., “Oleic Acid Protects Saturated Fatty Acid Mediated Lipotoxicity in Hepatocytes and Rat of Non-Alcoholic Steatohepatitis,” Life Sciences 203 (2018): 291–304, https://doi.org/10.1016/j.lfs.2018.04.022.

[101] F. Marra and G. Svegliati-Baroni, “Lipotoxicity and the Gut-Liver Axis in NASH Pathogenesis,” Journal of Hepatology 68, no. 2 (2018): 280–295, https://doi.org/10.1016/j.jhep.2017.11.014.

[102] W. Yang, R. Liu, C. Xia, et al., “Effects of Different Fatty Acids on BRL3A Rat Liver Cell Damage,” Journal of Cellular Physiology 235, no. 9 (2020): 6246–6256, https://doi.org/10.1002/jcp.29553.

[103] X. Palomer, J. Pizarro-Delgado, E. Barroso, and M. Vázquez-Carrera, “Palmitic and Oleic Acid: The Yin and Yang of Fatty Acids in Type 2 Diabetes Mellitus,” Trends in Endocrinology and Metabolism 29, no. 3 (2018): 178–190, https://doi.org/10.1016/j.tem.2017.11.009.

[104] L. L. Listenberger, X. Han, S. E. Lewis, et al., “Triglyceride Accumulation Protects Against Fatty Acid-Induced Lipotoxicity,” Proceedings of the National Academy of Sciences of the United States of America 100, no. 6 (2003): 3077–3082, https://doi.org/10.1073/pnas.0630588100.

[105] E. Obaseki, D. Adebayo, S. Bandyopadhyay, and H. Hariri, “Lipid Droplets and Fatty Acid-Induced Lipotoxicity: In a Nutshell,” FEBS Letters 598, no. 10 (2024): 1207–1214, https://doi.org/10.1002/1873-3468.14808.

[106] K. Maedler, G. A. Spinas, D. Dyntar, W. Moritz, N. Kaiser, and M. Y. Donath, “Distinct Effects of Saturated and Monounsaturated Fatty Acids on Beta-Cell Turnover and Function,” Diabetes 50, no. 1 (2001): 69–76, https://doi.org/10.2337/diabetes.50.1.69.

[107] Y. Wei, D. Wang, F. Topczewski, and M. J. Pagliassotti, “Saturated Fatty Acids Induce Endoplasmic Reticulum Stress and Apoptosis Independently of Ceramide in Liver Cells,” American Journal of Physiology. Endocrinology and Metabolism 291, no. 2 (2006): E275–E281, https://doi.org/10.1152/ajpendo.00644.2005.

[108] B. Zhang, W. Yang, S. Wang, et al., “Lipid Accumulation and Injury in Primary Calf Hepatocytes Challenged With Different Long-Chain Fatty Acids,” Frontiers in Veterinary Science 7 (2020): 547047, https://doi.org/10.3389/fvets.2020.547047.

[109] R. A. Igal, “Stearoyl-Coa Desaturase-1: A Novel Key Player in the Mechanisms of Cell Proliferation, Programmed Cell Death and Transformation to Cancer,” Carcinogenesis 31, no. 9 (2010): 1509–1515, https://doi.org/10.1093/carcin/bgq131.

[110] L. Liu, T. Shen, W. Yang, et al., “Ketotic Cows Display a Different Serum Nonesterified Fatty Acid Composition,” Journal of Dairy Research 87, no. 1 (2020): 52–55, https://doi.org/10.1017/S002202991900092X.

[111] E. A. Horst, S. K. Kvidera, and L. H. Baumgard, “Invited Review: The Influence of Immune Activation on Transition Cow Health and Performance-a Critical Evaluation of Traditional Dogmas,” Journal of Dairy Science 104, no. 8 (2021): 8380–8410, https://doi.org/10.3168/jds.2021-20330.

[112] E. Trevisi, M. Amadori, S. Cogrossi, E. Razzuoli, and G. Bertoni, “Metabolic Stress and Inflammatory Response in High-Yielding, Periparturient Dairy Cows,” Research in Veterinary Science 93, no. 2 (2012): 695–704, https://doi.org/10.1016/j.rvsc.2011.11.008.

[113] B. J. Bradford, L. K. Mamedova, J. E. Minton, J. S. Drouillard, and B. J. Johnson, “Daily Injection of Tumor Necrosis Factor-{Alpha} Increases Hepatic Triglycerides and Alters Transcript Abundance of Metabolic Genes in Lactating Dairy Cattle,” Journal of Nutrition 139, no. 8 (2009): 1451–1456, https://doi.org/10.3945/jn.109.108233.

[114] E. Galamb, V. Faigl, M. Keresztes, et al., “Effect of Pre- and Post-Partum Supplementation With Lipid-Encapsulated Conjugated Linoleic Acid on Milk Yield and Metabolic Status in Multiparous High-Producing Dairy Cows,” Journal of Animal Physiology and Animal Nutrition 101, no. 5 (2017): 1026–1035, https://doi.org/10.1111/jpn.12544.

[115] U. Moallem, H. Lehrer, L. Livshits, and M. Zachut, “The Effects of Omega-3 Α-Linolenic Acid From Flaxseed Oil Supplemented to High-Yielding Dairy Cows on Production, Health, and Fertility,” Livestock Science 242 (2020): 104302, https://doi.org/10.1016/j.livsci.2020.104302.

[116] A. Veshkini, F. Ceciliani, M. Bonnet, and H. M. Hammon, “Review: Effect of Essential Fatty Acids and Conjugated Linoleic Acid on the Adaptive Physiology of Dairy Cows During the Transition Period,” supplement, Animal 17, no. S2 (2023): 100757, https://doi.org/10.1016/j.animal.2023.100757.

[117] J. Fabjanowska, E. Kowalczuk-Vasilev, R. Klebaniuk, S. Milewski, and H. Gümüş, “N-3 Polyunsaturated Fatty Acids as a Nutritional Support of the Reproductive and Immune System of Cattle-a Review,” Animals (Basel) 13, no. 22 (2023): 3589, https://doi.org/10.3390/ani13223589.

[118] K. Libera, J. Wlodarek, E. Warzych, et al., “Reproductive Performance of Dairy Cows Fed a Diet Supplemented With N-3 Polyunsaturated Fatty Acids—A Review,” Annals of Animal Science 20, no. 4 (2020): 1169–1183, https://doi.org/10.2478/aoas-2020-0040.

[119] U. Moallem, “Invited Review: Roles of Dietary N-3 Fatty Acids in Performance, Milk Fat Composition, and Reproductive and Immune Systems in Dairy Cattle,” Journal of Dairy Science 101, no. 10 (2018): 8641–8661, https://doi.org/10.3168/jds.2018-14772.

[120] S. Lashkari, M. R. Weisbjerg, L. Foldager, and C. F. Børsting, “Fat Supplement for Dairy Cows During Early Lactation—Potentials, Challenges, and Risks—A Meta-Analysis,” Journal of Applied Animal Research 52, no. 1 (2024): 2323625, https://doi.org/10.1080/09712119.2024.2323625.

[121] R. M. Rodney, P. Celi, W. Scott, K. Breinhild, and I. Lean, “Effects of Dietary Fat on Fertility of Dairy Cattle: A Meta-Analysis and Meta-Regression,” Journal of Dairy Science 98, no. 8 (2015): 5601–5620, https://doi.org/10.3168/jds.2015-9528.

[122] M. Chirivi, D. Cortes-Beltran, A. Munsterman, A. O’Connor, and G. A. Contreras, “Lipolysis Inhibition as a Treatment of Clinical Ketosis in Dairy Cows: A Randomized Clinical Trial,” Journal of Dairy Science 106, no. 12 (2023): 9514–9531, https://doi.org/10.3168/jds.2023-23409.

[123] R. Schmitt, L. Pieper, S. Borchardt, J. Swinkels, C. C. Gelfert, and R. Staufenbiel, “Effects of a Single Transdermal Administration of Flunixin Meglumine in Early Postpartum Holstein Friesian Dairy Cows: Part 1. Inflammatory and Metabolic Markers, Uterine Health, and Indicators of Pain,” Journal of Dairy Science 106, no. 1 (2023): 624–640, https://doi.org/10.3168/jds.2021-20555.

[124] A. A. Barragan, E. Hovingh, S. Bas, et al., “Effects of Postpartum Acetylsalicylic Acid on Metabolic Status, Health, and Production in Lactating Dairy Cattle,” Journal of Dairy Science 103, no. 9 (2020): 8443–8452, https://doi.org/10.3168/jds.2019-17966.

[125] Z. Kovacevic, D. Stojanovic, M. Cincovic, et al., “Effect of Postpartum Administration of Ketoprofen on Proinflammatory Cytokine Concentration and Their Correlation With Lipogenesis and Ketogenesis in Holstein Dairy Cows,” Polish Journal of Veterinary Sciences 22, no. 3 (2019): 609–615, https://doi.org/10.24425/pjvs.2019.129970.

[126] O. B. Pascottini, S. J. Van Schyndel, J. F. W. Spricigo, et al., “Effect of Anti-Inflammatory Treatment on Systemic Inflammation, Immune Function, and Endometrial Health in Postpartum Dairy Cows,” Scientific Reports 10, no. 1 (2020): 5236, https://doi.org/10.1038/s41598-020-62103-x.

[127] A. J. Carpenter, C. M. Ylioja, C. F. Vargas, et al., “Hot Topic: Early Postpartum Treatment of Commercial Dairy Cows With Nonsteroidal Antiinflammatory Drugs Increases Whole-Lactation Milk Yield,” Journal of Dairy Science 99, no. 1 (2016): 672–679, https://doi.org/10.3168/jds.2015-10048.

[128] N. V. Priest, S. McDougall, C. R. Burke, et al., “The Responsiveness of Subclinical Endometritis to a Nonsteroidal Antiinflammatory Drug in Pasture-Grazed Dairy Cows,” Journal of Dairy Science 96, no. 7 (2013): 4323–4332, https://doi.org/10.3168/jds.2012-6266.

[129] S. Meier, N. V. Priest, C. R. Burke, et al., “Treatment With a Nonsteroidal Antiinflammatory Drug After Calving Did Not Improve Milk Production, Health, or Reproduction Parameters in Pasture-Grazed Dairy Cows,” Journal of Dairy Science 97, no. 5 (2014): 2932–2943, https://doi.org/10.3168/jds.2013-7838.

[130] K. Petrovic, D. Stojanovic, M. R. Cincovic, et al., “Influence of Niacin Application on Inflammatory Parameters, Non-Esterified Fatty Acids and Functional Status of Liver in Cows During Early Lactation,” Large Animal Review 27, no. 1 (2021): 17–21.

[131] A. Winkler, D. K. Gessner, C. Koch, et al., “Effects of a Plant Product Consisting of Green Tea and Curcuma Extract on Milk Production and the Expression of Hepatic Genes Involved in Endoplasmic Stress Response and Inflammation in Dairy Cows,” Archives of Animal Nutrition 69, no. 6 (2015): 425–441, https://doi.org/10.1080/1745039X.2015.1093873.

[132] M. V. Riboni, S. Meier, N. V. Priest, et al., “Adipose and Liver Gene Expression Profiles in Response to Treatment With a Nonsteroidal Antiinflammatory Drug After Calving in Grazing Dairy Cows,” Journal of Dairy Science 98, no. 5 (2015): 3079–3085, https://doi.org/10.3168/jds.2014-8579.

[133] M. Giammarco, I. Fusaro, G. Vignola, et al., “Effects of a Single Injection of Flunixin Meglumine or Carprofen Postpartum on Haematological Parameters, Productive Performance and Fertility of Dairy Cattle,” Animal Production Science 58, no. 2 (2018): 322–331, https://doi.org/10.1071/AN16028.

[134] N. C. Newby, D. L. Pearl, S. J. LeBlanc, K. E. Leslie, M. A. von Keyserlingk, and T. F. Duffield, “Effects of Meloxicam on Milk Production, Behavior, and Feed Intake in Dairy Cows Following Assisted Calving,” Journal of Dairy Science 96, no. 6 (2013): 3682–3688, https://doi.org/10.3168/jds.2012-6214.

RIGHTS & PERMISSIONS

2026 The Author(s). Animal Research and One Health published by John Wiley & Sons Australia, Ltd on behalf of Institute of Animal Science, Chinese Academy of Agricultural Sciences.