Effect of leptin on the gluconeogenesis in calf hepatocytes cultured in vitro

Liu, Yu; Li, Xiaobing; Song, Yuxiang; Shi, Xiaoxia; Ding, Hongyan; Yang, Wentao; Liu, Zhaoxi; Chen, Hui; Li, Xinwei; Wang, Jianguo; Liu, Guowen; Wang, Zhe

doi:10.1002/cbin.10172

Cited by 5 publications

(2 citation statements)

References 21 publications

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“…Neonatal calf was provided by the Experimental Animal Centre of Jilin University (Changchun, China) and the procedures were approved by the Institutional Animal Care and Use Committee of Jilin University (Changchun, China). Hepatocytes were collected and cultured as described in previous studies (Zhang et al 2012; Li et al 2013, 2015). The caudate lobe of the liver was obtained under sterile conditions from a female Holstein calf anesthetised with thiamylal sodium and was quickly placed on a sterile bench.…”

Section: Methodsmentioning

confidence: 99%

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

Ding

Wang

et al. 2016

Journal of Dairy Research

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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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Section: Methodsmentioning

confidence: 99%

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

Ding

Wang

et al. 2016

Journal of Dairy Research

Self Cite

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“…The process of obtaining vital hepatocytes from slaughterhouse-derived liver has already been described for equine (Stefanski et al, 2013[13]), pigs (Gerlach et al, 1993[2]) and recently for buffalo liver (Panda et al, 2015[9]). As mentioned above studies on bovine primary hepatocytes, by the majority obtain the hepatocytes from neonates or calves surgically and then euthanize the animals (Donkin and Armentano, 1993[1]; Li et al, 2013[7]; Zhang et al, 2016[17]). Donkin and Armentano (1993[1]) have described a process for the culturing of primary bovine hepatocytes up to 41 h after separation from male calves.…”

Section: Introductionmentioning

confidence: 99%