β-Hydroxybutyric Acid Inhibits Activation of Nuclear Factor-&#954;B Signaling Pathway in Lipopolysaccharide-Stimulated Neutrophils from Cows

ZHANG Yuming; YU Chunwei; DU Liyin; DENG Qinghua

doi:10.3969/j.issn.1006-267x.2022.02.058

2022 , Vol. 34 >Issue 2: 1268 - 1275

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.02.058

RESEARCH PAPER

β-Hydroxybutyric Acid Inhibits Activation of Nuclear Factor-κB Signaling Pathway in Lipopolysaccharide-Stimulated Neutrophils from Cows

ZHANG Yuming ,
YU Chunwei ,
DU Liyin ,
DENG Qinghua

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1. College of Animal Science and Technology, Inner Mongolia University for Nationalities, Tongliao 028000, China;
2. Inner Mongolia Agricultural University, Hohhot 010000, China

Received date: 2021-06-30

Online published: 2022-02-15

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Abstract

The innate immune function of polymorphonuclear neutrophils is inhibited in hyperketonemia cows. Therefore, this study was to investigate that β-hydroxybutyrate acid (BHBA) whether inhibited lipopolysaccharide (LPS)-induced activation of the nuclear factor-κB (NF-κB) signaling pathway in neutrophils from cows. The cow's neutrophils were isolated and treated by LPS (100 ng/mL) and different concentrations (0.5, 1.0, 2.0 and 4.0 mmol/L) of BHBA. Then the cells were collected. Real-time fluorescent quantitative PCR (qRT-PCR) was used to detect interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and NF-κBp65 mRNA expression levels in neutrophils; the protein expression level of NF-κBp65 in neutrophils was detected by Western blot; the inhibitor kappa B kinase β (IKKβ) kinase activity in neutrophils was measured by colorimetry; the secretion amounts of pro-inflammatory cytokines TNF-α, IL-6 and IL-1β in neutrophils were measured by enzyme-linked immunosorbent assay (ELISA) method. The results showed that compared with the control group (no BHBA and LPS treatment), the mRNA expression levels of IL-1β, IL-6, TNF-α and NF-κBp65 and the protein expression level of NF-κBp65 in the LPS group (only treated by LPS) were extremely significantly increased (P<0.01), the activity of IKKβ kinase was extremely significantly increased (P<0.01), the secretion amounts of IL-1β and TNF-α were extremely significantly increased (P<0.01), and the secretion amount of IL-6 was significantly increased (P<0.05). Compared with the LPS group, 0.5, 1.0, 2.0 and 4.0 mmol/L BHBA significantly or extremely significantly increased the mRNA expression levels of IL-1β, IL-6, TNF-α and NF-κBp65 and the protein expression level of NF-κBp65 (P<0.05 or P<0.01), significantly or extremely increased the activity of IKKβ kinase (P<0.05 or P<0.01), and significantly or extremely significantly increased the secretion amounts of IL-1β, IL-6 and TNF-α (P<0.05 or P<0.01). The above results indicate that BHBA can inhibit the activation of the NF-κB signaling pathway in LPS-induced neutrophils from cows, and has a certain anti-inflammatory function.

Key words： neutrophils; ketone body; inflammation; innate immunity

Cite this article

ZHANG Yuming , YU Chunwei , DU Liyin , DENG Qinghua . β-Hydroxybutyric Acid Inhibits Activation of Nuclear Factor-κB Signaling Pathway in Lipopolysaccharide-Stimulated Neutrophils from Cows[J]. Chinese Journal of Animal Nutrition, 2022 , 34(2) : 1268 -1275 . DOI: 10.3969/j.issn.1006-267x.2022.02.058

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