动物营养学报 >

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

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

研究论文 RESEARCH PAPER

β-羟丁酸抑制脂多糖诱导奶牛中性粒细胞核因子-κB信号通路的激活

  • 张玉明 ,
  • 于春微 ,
  • 杜立银 ,
  • 邓清华
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  • 1. 内蒙古民族大学动物科学技术学院, 通辽 028000;
    2. 内蒙古农业大学, 呼和浩特 010000
张玉明(1987-),男,山东聊城人,讲师,博士,主要从事兽医代谢免疫学研究。E-mail:zhangyuming1117@163.com

收稿日期: 2021-06-30

  网络出版日期: 2022-02-15

基金资助

内蒙古民族大学博士科研启动基金（BS476）

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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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摘要

高酮血症造成奶牛中性粒细胞先天免疫机能受到抑制，本研究探讨β-羟丁酸（BHBA）是否抑制脂多糖（LPS）诱导的奶牛中性粒细胞核因子-κB （NF-κB）信号通路的激活。分离健康奶牛中性粒细胞，采用LPS （100 ng/mL）和不同浓度（0.5、1.0、2.0和4.0 mmol/L） BHBA作用于中性粒细胞，收集细胞，应用实时荧光定量PCR （qRT-PCR）检测中性粒细胞中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和NF-κBp65 mRNA表达水平，Western blot检测NF-κBp65蛋白表达水平，比色法检测核因子-κB抑制物激酶β（IKKβ）激酶活性，酶联免疫吸附试验（ELISA）法检测促炎细胞因子TNF-α、IL-6和IL-1β的分泌量。结果表明：与对照组（不进行BHBA和LPS处理）相比较，LPS组（单独LPS处理）中IL-1β、IL-6、TNF-α和NF-κBp65 mRNA表达水平和NF-κBp65蛋白表达水平极显著增加（P<0.01），IKKβ激酶活性极显著增强（P<0.01），IL-1β和TNF-α的分泌量极显著增加（P<0.01），IL-6的分泌量显著增加（P<0.05）。与LPS组相比，0.5、1.0、2.0和4.0 mmol/L BHBA均使IL-1β、IL-6、TNF-α和NF-κBp65 mRNA表达水平和NF-κBp65蛋白表达水平显著或极显著增加（P<0.05或P<0.01），IKKβ激酶活性显著或极显著增强（P<0.05或P<0.01），IL-1β、IL-6和TNF-α的分泌量显著或极显著增加（P<0.05或P<0.01）。以上结果表明，高浓度BHBA可以抑制LPS诱导的奶牛中性粒细胞NF-κB信号通路的激活，具有一定的抗炎功能。

关键词： 中性粒细胞; 酮体; 炎症; 先天免疫

本文引用格式

张玉明 , 于春微 , 杜立银 , 邓清华 . β-羟丁酸抑制脂多糖诱导奶牛中性粒细胞核因子-κB信号通路的激活[J]. 动物营养学报, 2022 , 34(2) : 1268 -1275 . DOI: 10.3969/j.issn.1006-267x.2022.02.058

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

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