动物营养学报 >

2021 , Vol. 33 >Issue 9: 5246 - 5255

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.09.044

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

壳寡糖通过抑制白细胞介素-1β的生物学活性减缓由脂多糖诱导的奶牛外周血单个核细胞氧化应激损伤

  • 齐敬宇 ,
  • 郑亚光 ,
  • 郝颖 ,
  • 赵艳丽 ,
  • 郭晓宇 ,
  • 郭咏梅 ,
  • 史彬林 ,
  • 闫素梅
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  • 内蒙古农业大学动物科学学院, 内蒙古自治区高校动物营养与饲料科学重点实验室, 呼和浩特 010018
齐敬宇(1995-),女,内蒙古赤峰人,硕士研究生,从事动物营养与饲料科学研究。E-mail:15034956369@163.com

收稿日期: 2021-02-27

  网络出版日期: 2021-09-18

基金资助

国家自然科学基金项目（31671463）

收起

Chitosan Alleviates Lipopolysaccharide-Induced Oxidative Stress Damage in Peripheral Blood Mononuclear Cells of Dairy Cows by Inhibiting Interleukin-1β Biological Activity

  • QI Jingyu ,
  • ZHENG Yaguang ,
  • HAO Ying ,
  • ZHAO Yanli ,
  • GUO Xiaoyu ,
  • GUO Yongmei ,
  • SHI Binlin ,
  • YAN Sumei
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  • College of Animal Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2021-02-27

  Online published: 2021-09-18

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摘要

本试验以脂多糖（LPS）为刺激源，利用白细胞介素-1受体拮抗剂（IL-1ra）阻断白细胞介素-1β（IL-1β）的生物学活性，探讨壳寡糖（COS）对LPS诱导的奶牛外周血单个核细胞（PBMC）氧化应激损伤的减缓作用。采用单因素完全随机试验设计，将PBMC随机分为8个组，分别为对照组（在不含COS、LPS和IL-1ra的基础培养基中培养72 h）、COS组（在仅含COS的培养基中培养72 h）、LPS组（在基础培养基中先培养48 h，再加入LPS培养24 h）、IL-1ra组（在基础培养基中先培养42 h，再加入IL-1ra培养30 h）、COS+IL-1ra组（在含有COS的培养基中先培养42 h，再加入IL-1ra培养30 h）、COS+LPS组（在含有COS的培养基中先培养48 h，再加入LPS培养24 h）、IL-1ra+LPS组（在基础培养基中先培养42 h，然后加入IL-1ra再培养6 h，最后加入LPS再培养24 h）和COS+IL-1ra+LPS组（在含有COS的培养基中先培养42 h，然后加入IL-1ra再培养6 h，最后加入LPS再培养24 h），每组6个重复。结果显示：与对照组相比，LPS组抗氧化酶谷胱甘肽过氧化物酶（GPx）、总超氧化物歧化酶（T-SOD）、过氧化氢酶（CAT）和硫氧化蛋白还原酶（TrxR）的活性与总抗氧化能力（T-AOC）以及GPx1和TrxR的mRNA相对表达量显著下降（P ≤ 0.05），而诱导型一氧化氮合酶（iNOS）活性，炎症因子IL-1β、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的含量及其mRNA相对表达量，核因子-κB（NF-κB）p65的mRNA相对表达量、一氧化氮（NO）与丙二醛（MDA）含量、活性氧簇（ROS）活性均显著升高（P ≤ 0.05），说明LPS诱导的奶牛PBMC氧化应激损伤是由于其激活了NF-κB信号通路，导致炎症因子IL-1β的释放以及iNOS活性和NO含量的增加造成的。与LPS组相比，LPS+IL-1ra组和LPS+COS组的上述抗氧化酶活性及GPx1和TrxR的mRNA相对表达量显著升高（P ≤ 0.05），而上述炎症因子的含量及其mRNA相对表达量以及NF-κB p65的mRNA相对表达量显著降低（P ≤ 0.05），说明IL-1ra和COS对LPS诱导的奶牛PBMC氧化应激损伤具有相似的预防作用。综上可知，COS通过抑制IL-1β的生物学活性减缓LPS诱导的奶牛PBMC氧化应激损伤，这可能与COS抑制NF-κB信号通路的活性，进而降低IL-1β的释放及NO的生成有关。

关键词： 壳聚糖; 奶牛外周血单个核细胞; 氧化应激; 白细胞介素-1受体拮抗剂

本文引用格式

齐敬宇 , 郑亚光 , 郝颖 , 赵艳丽 , 郭晓宇 , 郭咏梅 , 史彬林 , 闫素梅 . 壳寡糖通过抑制白细胞介素-1β的生物学活性减缓由脂多糖诱导的奶牛外周血单个核细胞氧化应激损伤[J]. 动物营养学报, 2021 , 33(9) : 5246 -5255 . DOI: 10.3969/j.issn.1006-267x.2021.09.044

Abstract

In this experiment, lipopolysaccharide (LPS) was used as the stimulation source, using interleukin-1 receptor antagonist (IL-1ra) to block the biological activity of interleukin-1β (IL-1β), to investigate the mitigating effect of chitosan (COS) on LPS-induced oxidative stress damage in peripheral blood mononuclear cells (PBMC) of dairy cows. With a one-way completely randomized trial design, PBMC were randomly divided into 8 groups, namely, control group (cultured in a basic medium without COS, LPS and IL-1Ra for 72 h), COS group (cultured in a medium with COS only for 72 h), LPS group (cultured in the basic medium for 48 h, and then added LPS for another 24 h), IL-1ra group (cultured in the basic medium for 42 h, and then added IL-1ra for another 30 h), COS+IL-1ra group (cultured in the medium with COS for 42 h, and then added IL-1ra for another 30 h), COS+LPS group (cultured in the medium with COS for 48 h, and then added LPS for another 24 h), IL-1ra+LPS group (cultured in the basic medium for 42 h, and then added IL-1ra for another 6 h, then added LPS for another 24 h), and COS+IL-1ra+LPS group (cultured in the medium with COS for 42 h, and then added IL-1ra for another 6 h, then added LPS for another 24 h), with 6 replicates in each group. The results showed that the activities of antioxidant enzymes glutathione peroxidase (GPx), total superoxide dismutase (T-SOD), catalase (CAT) and thioredoxin reductase (TrxR), the total antioxidant capacity (T-AOC) and the mRNA relative expression levels of GPx1 and TrxR were significantly decreased in the LPS group compared with the control group (P ≤ 0.05), while the activity of inducible nitric oxide synthase (iNOS), the contents of inflammatory factors IL-1β, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and their mRNA relative expression levels, the mRNA relative expression level of nuclear factor-κB (NF-κB) p65, the contents of nitric oxide (NO) and malondialdehyde (MDA), and the reactive oxygen species (ROS) activity were significantly increased (P ≤ 0.05), it indicated that LPS-induced oxidative stress damage in PBMC of dairy cows was caused by its activation of NF-κB signaling pathway, leading to the release of inflammatory factor IL-1β and an increase in iNOS activity and NO content. Compared with the LPS group, the above antioxidant enzyme activities and the mRNA relative expression levels of GPx1 and TrxR were significantly increased in the LPS+IL-1ra and LPS+COS groups (P ≤ 0.05), whereas the contents of the above inflammatory factors and their mRNA relative expression levels, as well as the mRNA relative expression level of NF-κB p65 were significantly decreased (P ≤ 0.05), it indicated that IL-1ra and COS had similar preventive effects on LPS-induced oxidative stress damage in PBMC of dairy cows. In summary, it is clear that COS mitigates LPS-induced oxidative stress damage in PBMC of dairy cows by inhibiting the biological activity of IL-1β, which may be related to COS inhibiting the activity of NF-κB signaling pathway and thus reducing the release of IL-1β and the production of NO.

Key words： chitosan; PBMC; oxidative stress; interleukin-1 receptor antagonist

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