动物营养学报 >

2020 , Vol. 32 >Issue 12: 5903 - 5909

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.12.043

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

茶树油对脂多糖诱导的奶牛瘤胃上皮细胞炎症因子表达的影响

  • 马晓宇 ,
  • 彭程 ,
  • 詹康 ,
  • 占今舜 ,
  • 杨天宇 ,
  • 宁丽丽 ,
  • 赵国琦
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  • 1. 扬州大学动物科学与技术学院, 扬州 225009;
    2. 扬州大学农业科技发展研究院, 扬州 225009;
    3. 扬州大学教育部农业与农产品安全国际合作联合实验室, 扬州 225009;
    4. 江西省农业科学院畜牧兽医研究所, 南昌 330200
马晓宇(1997-),女,山东潍坊人,硕士研究生,从事反刍动物营养、生产与环境研究。E-mail:maxiaoyu0417@163.com

收稿日期: 2020-05-24

  网络出版日期: 2020-12-07

基金资助

江苏省自然科学基金（SBK2019043455）；国家自然科学基金项目（31972589）；江西现代农业科研协同创新专项（JXXTCX201702-04）

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Effects of Tea Tree Oil on Expression of Inflammatory Factors in Bovine Rumen Epithelial Cells Induced by Lipopolysaccharide

  • MA Xiaoyu ,
  • PENG Cheng ,
  • ZHAN Kang ,
  • ZHAN Jinshun ,
  • YANG Tianyu ,
  • NING Lili ,
  • ZHAO Guoqi
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  • 1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
    2. Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China;
    3. Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University Yangzhou 225009, China;
    4. Institute of Animal Husbandry and Veterinary, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China

Received date: 2020-05-24

  Online published: 2020-12-07

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摘要

本试验旨在研究茶树油（TTO）对脂多糖（LPS）诱导的奶牛瘤胃上皮细胞（BRECs）炎症因子表达的影响。利用CCK-8法测定浓度为0（对照）、0.006 25%、0.012 5%、0.025%、0.05%、0.1%的TTO对BRECs活力的影响，每组6个重复，根据试验结果筛选出TTO的适宜浓度为0.05%，以此浓度做后续试验。再利用实时荧光定量PCR、蛋白质印迹法，分别探究BRECs中炎症因子在基因水平以及蛋白水平表达量的变化。试验分为4组：对照组不添加TTO和LPS；TTO组仅添加0.05%的TTO；LPS组仅添加1 μg/mL LPS；LPS+TTO组添加1 μg/mL LPS和0.05% TTO，每组6个重复。结果表明：1）与对照组相比，添加0.05% TTO显著提高BRECs的活力（P<0.05）；添加0.1% TTO显著抑制BRECs的活力（P<0.05）。2）与对照组相比，TTO组白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和Toll样受体-4（TLR-4）的基因表达量均显著降低（P<0.05），LPS组白细胞介素-1β（IL-1β）、IL-6、白细胞介素-8（IL-8）、TNF-α、TLR-4基因表达量显著升高（P<0.05）；与LPS组相比，在LPS中添加TTO显著降低IL-1β、IL-6、IL-8、TNF-α、TLR-4的基因表达量（P<0.05）。3）与对照组相比，LPS可以提高核因子-κB（NF-κB）通路中p65、磷酸化-p65（p-p65）蛋白表达量，添加LPS+TTO与仅添加LPS相比可以降低p65、p-p65蛋白表达量。综上所述，低浓度的TTO可以提高BRECs的活力，抑制LPS诱导的BRECs炎症信号通路NF-κB相关蛋白表达，进而抑制促炎因子产生。

关键词： 茶树油; 瘤胃上皮细胞; NF-&kappa; B; 促炎症因子

本文引用格式

马晓宇 , 彭程 , 詹康 , 占今舜 , 杨天宇 , 宁丽丽 , 赵国琦 . 茶树油对脂多糖诱导的奶牛瘤胃上皮细胞炎症因子表达的影响[J]. 动物营养学报, 2020 , 32(12) : 5903 -5909 . DOI: 10.3969/j.issn.1006-267x.2020.12.043

Abstract

The purpose of this study was to investigate the effects of tea tree oil (TTO) on the expression of inflammatory factors in bovine rumen epithelial cells (BRECs) induced by lipopolysaccharide (LPS). The cell counting kit-8 (CCK-8) was used to determine the effects of different TTO concentrations [0 (control), 0.006 25%, 0.012 5%, 0.025%, 0.05% and 0.1% TTO] on the activity of BRECs and each concentration was performed six replicates. The concentration of 0.05% TTO was selected according to the above tests for subsequent experiments. Then, qRT-PCR and Western blot were used to investigate the changes of expression levels of inflammatory factors at gene and protein levels in BRECs. The experiment was divided into four groups: control group (CON group, without adding TTO and LPS), TTO group (only adding 0.05% TTO), LPS group (only adding 1 μg/mL LPS), LPS+TTO group (both adding 1 μg/mL LPS and 0.05% TTO). Each group was performed six replicates. The results showed as follows: 1) compared with the CON group, the cell viability of BRECs was significantly improved by adding 0.05% TTO (P<0.05) and was significantly inhibited by adding 0.1% TTO (P<0.05). 2) Compared with the CON group, the expression levels of interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α (TNF-α) and Toll like receptor-4 (TLR-4) genes were significantly increased when LPS was added (P<0.05). Compared with LPS group, the expression levels of IL-1β, IL-6, IL-8, TNF-α and TLR-4 genes were significantly decreased by adding TTO to LPS (P<0.05). 3) Compared with the CON group, LPS could increase the protein expression levels of p65 and p-p65 in nuclear factor-κB (NF-κB) pathway but adding both LPS and TTO could decrease the protein expression levels of p65 and p-p65. In summary, low concentration of TTO can enhance the activity of BRECs and inhibit the expression of proteins associated with inflammatory signaling pathway NF-κB induced by inflammation of BRECs which caused by LPS, and then can inhibit the production of proinflammatory factor.

Key words： tea tree oil; bovine rumen epithelial cells; NF-κB; proinflammatory cytokine

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