动物营养学报 >

2021 , Vol. 33 >Issue 8: 4637 - 4644

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.08.043

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

褪黑素对脂多糖诱导的奶牛乳腺上皮细胞炎症反应的缓解作用

  • 李洪洋 ,
  • 南雪梅 ,
  • 孙鹏
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  • 中国农业科学院北京畜牧兽医研究所, 动物营养学国家重点实验室, 北京 100193
李洪洋(1995-),女,辽宁沈阳人,硕士研究生,从事动物营养与饲料科学研究。E-mail:929307601@qq.com

收稿日期: 2021-01-11

  网络出版日期: 2021-08-11

基金资助

国家重点研发计划（2016YFD0500507）；中国农业科学院科技创新工程（ASTIP-IAS07）

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Mitigative Effects of Melatonin on Lipopolysaccharide-Induced Inflammation in Bovine Mammary Epithelial Cells

  • LI Hongyang ,
  • NAN Xuemei ,
  • SUN Peng
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  • State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received date: 2021-01-11

  Online published: 2021-08-11

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摘要

本试验旨在探讨褪黑素（MT）对脂多糖（LPS）诱导的奶牛乳腺上皮细胞（BMECs）炎症反应的缓解作用及潜在机制。利用不同浓度（0.1、0.5、1.0、5.0和10.0 μg/mL）的LPS处理BMECs 6和12 h后，通过测定BMECs活性和炎性细胞因子[肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）]含量，确定10 μg/mL的LPS诱导12 h用于正式试验。正式试验共设7组，每组设4个重复。对照组BMECs不进行LPS诱导和MT处理，LPS组BMECs进行LPS诱导12 h，LPS+MT组BMECs进行LPS诱导12 h后再经不同浓度（1、5、10、50和100 μmol/L） MT处理48 h。结果表明：1）与对照组相比，LPS组BMECs中TNF-α含量升高（P>0.05），BMECs中IL-6和IL-1β含量显著升高（P<0.05）。与LPS组相比，10和50 μmol/L MT组BMECs中TNF-α含量显著降低（P<0.05），1、5、10和100 μmol/L MT组BMECs中IL-6含量显著降低（P<0.05），10 μmol/L MT组BMECs中IL-1β含量显著降低（P<0.05）。2）与对照组相比，LPS组BMECs中Toll样受体4（TLR4）蛋白表达量升高（P>0.05），BMECs中核因子-κB同源蛋白（P65）和核因子-κB抑制蛋白-α（IκB-α）蛋白表达量显著降低（P<0.05）。与LPS组相比，50和100 μmol/L MT组BMECs中TLR4蛋白表达量显著降低（P<0.05），1、10和50 μmol/L MT组BMECs中P65蛋白表达量显著或极显著升高（P<0.05或P<0.01），1、5、10和50 μmol/L MT组BMECs中IκB-α蛋白表达量显著或极显著升高（P<0.05或P<0.01）。由此可见，MT能够降低LPS诱导的BMECs中TNF-α、IL-1β和IL-6含量，调控LPS诱导的BMECs中TLR4、P65和IκB-α蛋白表达量，MT可能通过参与核因子-κB炎症通路缓解LPS诱导的BMECs炎症反应。

关键词： 乳房炎; 褪黑素; 脂多糖; 抗炎; 炎症通路

本文引用格式

李洪洋 , 南雪梅 , 孙鹏 . 褪黑素对脂多糖诱导的奶牛乳腺上皮细胞炎症反应的缓解作用[J]. 动物营养学报, 2021 , 33(8) : 4637 -4644 . DOI: 10.3969/j.issn.1006-267x.2021.08.043

Abstract

The purpose of this study was to investigate the mitigative effects and underlying mechanisms of melatonin (MT) on lipopolysaccharide (LPS)-induced inflammation in bovine mammary epithelial cells (BMECs). The BMECs were treated with different concentrations (0.1, 0.5, 1.0, 5.0 and 10.0 μg/mL) of LPS for 6 and 12 hours, respectively. The BMECs viability was detected and the inflammatory cytokines[tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6)] contents was measured, and the BMECs model was established with 12 hours and 10 μg/mL LPS for formal experiment. The formal experiment divided into 7 groups with 4 replicates in each group. BMECs in the control group without LPS-induced and MT treated, BMECs in the LPS group were LPS-induced 12 hours, and BMECs in LPS+MT groups were LPS-induced 12 hours and treated with different concentrations (1, 5, 10, 50 and 100 μmol/L) of LPS for 48 hours, respectively. The results showed as follows:1) compared with the control group, the BMECs TNF-α content of LPS group was increased (P>0.05), and the contents of IL-6 and IL-1β in BMECs were significantly increased (P<0.05). Compared with the LPS group, the BMECs TNF-α content of 10 and 50 μmol/L MT groups was significantly decreased (P<0.05), the BMECs IL-6 content of 1, 5, 10 and 100 μmol/L MT groups was significantly decreased (P<0.05), and the BMECs IL-1β content of 10 μmol/L MT group was significantly decreased (P<0.05). 2) Compared with the control group, the BMECs Toll-like receptor 4 (TLR4) protein expression level of LPS group was increased (P>0.05), and the protein expression levels of nuclear factor-κB homologous protein (P65) and nuclear factor-κB inhibitory protein-α (IκB-α) in BMECs were significantly decreased (P<0.05). Compared with the LPS group, the BMECs TLR4 protein expression level of 10 and 100 μmol/L MT groups was significantly decreased (P<0.05), the BMECs P65 protein expression level of 1, 10 and 50 μmol/L MT groups was significantly increased (P<0.05 or P<0.01), and the BMECs IκB-α protein expression level of 1, 5, 10 and 50 μmol/L MT group was significantly increased (P<0.05 or P<0.01). It is concluded that MT can reduce the contents of TNF-α, IL-1β and IL-6 in BMECs induced by LPS, regulate the protein expression levels of TLR4, P65 and IκB-α in BMECs induced by LPS, and likely participate nuclear factor-κB inflammatory pathway, which exerted anti-inflammatory effect in LPS-challenged BMECs.

Key words： mastitis; melatonin; lipopolysaccharide; anti-inflammatory; inflammatory pathway

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