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High levels of acetoacetate and glucose increase expression of cytokines in bovine hepatocytes, through activation of the NF-κB signalling pathway

  • Yu Li (a1), Hongyan Ding (a1), Xichun Wang (a1), Lei Liu (a2), Dan Huang (a2), Renhe Zhang (a2), Lihui Guo (a2), Zhe Wang (a2), Xiaobing Li (a2), Guowen Liu (a2), Jinjie Wu (a1) and Xinwei Li (a2)...


Elevated levels of blood interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumour necrosis factor–α (TNF–α) increase insulin resistance and result in inflammation. It is not clear whether elevated blood level of acetoacetate (ACAC) and decreased blood level of glucose, which are the predominant characteristics of clinical biochemistry in ketotic dairy cows, increase proinflammatory cytokines and subsequent inflammation. The objective of this study was to test the hypothesis that ACAC and glucose activate the NF-κB signalling pathway to regulate cytokines expression in bovine hepatocytes. Bovine hepatocytes were cultured with ACAC (0–4·8 mm) and glucose (0–5·55 mm) with or without NF-κB inhibitor PDTC for 24 h. The secretion and mRNA levels of cytokines were determined by enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The NF-κB signalling pathway activation was evaluated by western blotting. Results showed that the secretion and expression of IL-1β, IL-6 and TNF-α increased in an ACAC dose-dependent manner. Additionally, there was an increase in the secretion and mRNA expression of these three cytokines in glucose treatment group, which increased significantly when the glucose concentrations exceed 3·33 mm. Furthermore, both ACAC and glucose upregulated NF-κB p65 protein expression and IκBα phosphorylation levels. However, these effects were reduced by PDTC. These results demonstrate that elevated levels of ACAC and glucose increase the synthesis and expression of proinflammatory factors by activating NF-κB signalling pathway in hepatocytes, which may contribute to inflammation injury in ketotic dairy cows.


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Bannerman, DD, Paape, MJ, Lee, JW, Zhao, X, Hope, JC & Rainard, P 2004 Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clinical and Diagnostic Laboratory Immunology 11 463472
Bertoni, G, Trevisi, E, Han, X & Bionaz, M 2008 Effects of inflammatory conditions on liver activity in puerperium period and consequences for performance in dairy cows. Journal of Dairy Science 91 33003310
Bruun, JM, Helge, JW, Richelsen, B & Stallknecht, B 2006 Diet and exercise reduce low-grade inflammation and macrophage infiltration in adipose tissue but not in skeletal muscle in severely obese subjects. American Journal of Physiology-Endocrinology and Metabolism 290 961967
Dasarathy, S 2008 Inflammation and liver. Journal of Parenteral and Enteral Nutrition 32 660666
Dasu, MR, Devaraj, S, Zhao, L, Hwang, DH & Jialal, I 2008 High glucose induces Toll-Like receptor expression in human monocytes. Diabetes 57 30903098
Drackley, JK, Dann, HM, Douglas, GN, Guretzky, NAJ, Litherland, NB, Underwood, JP & Loor, JJ 2005 Physiological and pathological adaptations in dairy cows that may increase susceptibility to periparturient diseases and disorders. Italian Journal of Animal Science 4 323344
Gerszten, RE, Garcia-Zepeda, EA, Lim, YC, Yoshida, M, Ding, HA, Gimbrone, MA, Luster, AD, Luscinskas, FW & Rosenzweig, A 1999 MCP-1 and IL-8 trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Nature 398 718723
Grummer, RR 1995 Impact of changes in organic nutrient metabolism on feeding the transition dairy cow. Journal of Animal Science 73 28202820
Hichem, L, Etienne, M, Pascal, R & Céline, R 2007 Differential cytokine and chemokine responses of bovine mammary epithelial cells to Staphylococcus aureus and Escherichia coli. Cytokine 38 1221
Hoffman, WH, Cheng, C, Passmore, GG, Carroll, JE & Hess, D 2002 Acetoacetate increases expression of intercellular adhesion molecule-1 (ICAM-1) in human brain microvascular endothelial cells. Neuroscience Letters 334 7174
Jain, SK, Rains, JL & Croad, JL 2007 High glucose and ketosis (acetoacetate) increases, and chromium niacinate decreases, IL-6, IL-8, and MCP-1 secretion and oxidative stress in U937 monocytes. Antioxidants & Redox Signaling 9 15811590
Kremer, WDJ, Noordhuizen-Stassen, EN, Grommers, FJ, Schukken, YH, Heeringa, R, Brand, A & Burvenich, C 1993 Severity of experimental Escherichia coli mastitis in ketonemic and nonketonemic dairy cows. Journal of Dairy Science 76 34283436
LeBlanc, S 2010 Monitoring metabolic health of dairy cattle in the transition period. Journal of Reproduction and Development 56(S) S29S35
Li, X, Li, X, Bai, G, Chen, H, Deng, Q, Liu, Z, Zhang, L, Liu, G & Wang, Z 2012 Effects of non-esterified fatty acids on the gluconeogenesis in bovine hepatocytes. Molecular and Cellular Biochemistry 359 385388
Li, X, Li, Y, Yang, W, Xiao, C, Fu, S, Deng, Q, Ding, H, Wang, Z, Liu, G & Li, X 2014 SREBP-1c overexpression induces triglycerides accumulation through increasing lipid synthesis and decreasing lipid oxidation and VLDL assembly in bovine hepatocytes. Journal of Steroid Biochemistry & Molecular Biology 143 174182
Li, X, Huang, W, Gu, J, Du, X, Lei, L, Yuan, X, Sun, G, Wang, Z, Li, X & Liu, G 2015 SREBP-1c overactivates ROS-mediated hepatic NF-κB inflammatory pathway in dairy cows with fatty liver. Cellular Signalling 27 20992109
Li, Y, Li, X, Song, Y, Shi, X, Ding, H, Yang, W, Liu, Z, Chen, H, Li, X & Wang, J 2013 Effect of leptin on the gluconeogenesis in calf hepatocytes cultured in vitro. Cell Biology International 37 13501353
Lin, Y, Berg, AH, Iyengar, P, Lam, TKT, Giacca, A, Combs, TP, Rajala, MW, Du, X, Rollman, B & Li, W 2005 The hyperglycemia-induced inflammatory response in adipocytes the role of reactive oxygen species. Journal of Biological Chemistry 280 46174626
Manco, M, Marcellini, M, Giannone, G & Nobili, V 2007 Correlation of serum TNF-alpha levels and histologic liver injury scores in pediatric nonalcoholic fatty liver disease. American Journal of Clinical Pathology 127 954960
Ohtsuka, H, Koiwa, M, Hatsugaya, A, Kudo, K, Hoshi, F, Itoh, N, Yokota, H, Okada, H & Kawamura, S-i 2001 Relationship between serum TNF activity and insulin resistance in dairy cows affected with naturally occurring fatty liver. Journal of Veterinary Medical Science 63 10211025
Park, SW, Chen, SWC, Kim, M, Brown, KM, Kolls, JK, D D'Agati, V & Lee, HT 2011 Cytokines induce small intestine and liver injury after renal ischemia or nephrectomy. Laboratory Investigation 91 6384
Shanmugam, N, Reddy, MA, Guha, M & Natarajan, R 2003 High glucose-induced expression of proinflammatory cytokine and chemokine genes in monocytic cells. Diabetes 52 12561264
Shi, X, Li, X, Li, D, Li, Y, Song, Y, Deng, Q, Wang, J, Zhang, Y, Ding, H & Yin, L 2014 β-Hydroxybutyrate activates the NF-κB signaling pathway to promote the expression of pro-inflammatory factors in calf hepatocytes. Cellular Physiology and Biochemistry: International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology 33 920932
Shi, X, Li, D, Deng, Q, Li, Y, Sun, G, Yuan, X, Song, Y, Wang, Z, Li, X & Li, X 2015 NEFAs activate the oxidative stress-mediated NF-κB signaling pathway to induce inflammatory response in calf hepatocytes. The Journal of Steroid Biochemistry and Molecular Biology 145 103112
Stentz, FB, Umpierrez, GE, Cuervo, R & Kitabchi, AE 2004 Proinflammatory cytokines, markers of cardiovascular risks, oxidative stress, and lipid peroxidation in patients with hyperglycemic crises. Diabetes 53 20792086
Targowski, SP & Klucinski, W 1983 Reduction in mitogenic response of bovine lymphocytes by ketone bodies. American Journal of Veterinary Research 44 828830
Wang, X, Li, X, Zhao, C, Hu, P, Chen, H, Liu, Z, Liu, G & Wang, Z 2012 Correlation between composition of the bacterial community and concentration of volatile fatty acids in the rumen during the transition period and ketosis in dairy cows. Applied and Environmental Microbiology 78 23862392
Wathes, DC, Cheng, Z, Fenwick, MA, Fitzpatrick, R & Patton, J 2011 Influence of energy balance on the somatotrophic axis and matrix metalloproteinase expression in the endometrium of the postpartum dairy cow. Reproduction 141 269281
Xu, C, Liu, GW, Li, XB, Xia, C, Zhang, HY & Wang, Z 2010 Decreased complete oxidation capacity of fatty acid in the liver of ketotic cows. Asian Australasian Journal of Animal Science 3 312317
Zdzisińska, B, Filar, J, Paduch, R, Kaczor, J, Lokaj, I & Kandefer-Szerszeń, M 2000 The influence of ketone bodies and glucose on interferon, tumor necrosis factor production and NO release in bovine aorta endothelial cells. Veterinary Immunology and Immunopathology 74 237247
Zhang, ZG, Li, XB, Gao, L, Liu, GW, Kong, T, Li, YF, Wang, HB, Zhang, C, Wang, Z & Zhang, R-H 2012 An updated method for the isolation and culture of primary calf hepatocytes. The Veterinary Journal 191 323326
Zhang, ZG, Xue, JD, Gao, RF, Liu, JY, Wang, JG, Yao, CY, Liu, Y, Li, XW, Li, XB & Liu, GW 2013 Evaluation of the difference of L-selectin, tumor necrosis factor-α and sialic acid concentration in dairy cows with subclinical ketosis and without subclinical ketosis. Pakistan Veterinary Journal 33 225228


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High levels of acetoacetate and glucose increase expression of cytokines in bovine hepatocytes, through activation of the NF-κB signalling pathway

  • Yu Li (a1), Hongyan Ding (a1), Xichun Wang (a1), Lei Liu (a2), Dan Huang (a2), Renhe Zhang (a2), Lihui Guo (a2), Zhe Wang (a2), Xiaobing Li (a2), Guowen Liu (a2), Jinjie Wu (a1) and Xinwei Li (a2)...


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