title: High levels of acetoacetate and glucose increase expression of cytokines in bovine hepatocytes, through activation of the NF-κB signalling pathway.
abstract: 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.
authors: College of Animal Science and Technology,Anhui Agricultural University,130 West Changjiang Road,Hefei 230036,China.
authors: Yu
authors: Y
authors: Li
authors: College of Animal Science and Technology,Anhui Agricultural University,130 West Changjiang Road,Hefei 230036,China.
authors: Hongyan
authors: H
authors: Ding
authors: College of Animal Science and Technology,Anhui Agricultural University,130 West Changjiang Road,Hefei 230036,China.
authors: Xichun
authors: X
authors: Wang
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Lei
authors: L
authors: Liu
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Dan
authors: D
authors: Huang
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Renhe
authors: R
authors: Zhang
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Lihui
authors: L
authors: Guo
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Zhe
authors: Z
authors: Wang
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Xiaobing
authors: X
authors: Li
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Guowen
authors: G
authors: Liu
authors: College of Animal Science and Technology,Anhui Agricultural University,130 West Changjiang Road,Hefei 230036,China.
authors: Jinjie
authors: J
authors: Wu
authors: College of Veterinary Medicine,Jilin University,5333 Xi'an Road,Changchun,Jilin 130062,China.
authors: Xinwei
authors: X
authors: Li
mesh_terms: D000090:Acetoacetates / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013859:Thiocarbamates / Q000494:pharmacology
chemical_list: D000090:Acetoacetates; D016207:Cytokines; D016328:NF-kappa B; D012333:RNA, Messenger; D013859:Thiocarbamates; C066229:prolinedithiocarbamate; C016635:acetoacetic acid; D011392:Proline; D005947:Glucose
keywords: ACAC; NF-κB; bovine hepatocytes; glucose; proinflammatory cytokines