研究论文 RESEARCH PAPER

大豆活性肽对体外过氧化氢诱导的奶牛乳腺上皮细胞氧化应激和炎症损伤的缓解作用

  • 陈逸 ,
  • 张一涵 ,
  • 张静 ,
  • 屠焰 ,
  • 王慧 ,
  • 蒋林树
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  • 1. 北京农学院动物科学技术学院, 奶牛营养学北京市重点实验室, 北京 102206;
    2. 中国农业科学院饲料研究所, 北京 100081
陈逸(1998-),女,江苏盐城人,硕士研究生,研究方向为反刍动物营养与免疫。E-mail:luckymelon0826@163.com

收稿日期: 2022-04-13

  网络出版日期: 2022-10-17

基金资助

北京市教育委员会重点项目(KZ202010020029);国家自然科学基金面上项目(31772629)

Alleviation Effect of Soybean Bioactive Peptide on Oxidative Stress and Inflammatory Injury of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide in Vitro

  • CHEN Yi ,
  • ZHANG Yihan ,
  • ZHANG Jing ,
  • TU Yan ,
  • WANG Hui ,
  • JIANG Linshu
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  • 1. College of Animal Science and Technology, Beijing University of Agricultural, Beijing 102206, China;
    2. Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received date: 2022-04-13

  Online published: 2022-10-17

摘要

本试验旨在探究大豆活性肽(SBP)对体外过氧化氢(H2O2)诱导的奶牛乳腺上皮细胞(MAC-T)氧化应激和炎症损伤的缓解作用。试验采用体外细胞培养方法,以不同浓度H2O2处理MAC-T细胞建立细胞氧化应激模型,在此基础上以不同浓度(0.15、0.30、0.60、1.20、2.40、4.80 mg/mL) SBP对细胞进行预处理,测定细胞存活率以及细胞内活性氧(ROS)水平、抗氧化指标及炎症相关基因的mRNA相对表达水平,试验同时设置对照组和H2O2模型组。结果显示:与对照组相比, 600 μmol/L的H2O2作用于细胞12 h,细胞内ROS水平极显著升高(P<0.01),总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性和总抗氧化能力(T-AOC)极显著下降(P<0.01),丙二醛(MDA)含量极显著升高(P<0.01)。而以0.15、0.30和0.60 mg/mL SBP对细胞进行预处理后,与H2O2模型组相比,极显著降低了细胞内ROS水平(P<0.01),极显著提高了T-SOD、GSH-Px、CAT活性(P<0.01),显著或极显著提高了T-AOC(P<0.05或P < 0.01),极显著降低了MDA含量(P<0.01),极显著降低了炎症介质[环氧化酶-2(COX-2)、诱导型一氧化氮合酶(iNOS)、单核细胞趋化蛋白-1(MCP-1)]和促炎因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)]的mRNA相对表达水平(P<0.01)。综上所述, SBP在体外可能通过降低细胞内ROS水平,提高细胞抗氧化酶活性,调节炎症相关基因的表达,来缓解H2O2诱导的MAC-T细胞的氧化应激和炎症损伤。

本文引用格式

陈逸 , 张一涵 , 张静 , 屠焰 , 王慧 , 蒋林树 . 大豆活性肽对体外过氧化氢诱导的奶牛乳腺上皮细胞氧化应激和炎症损伤的缓解作用[J]. 动物营养学报, 2022 , 34(10) : 6714 -6725 . DOI: 10.3969/j.issn.1006-267x.2022.10.064

Abstract

This experiment was conducted to investigate the alleviation effect of soybean bioactive peptide (SBP) on oxidative stress and inflammatory injury of bovine mammary epithelial cells (MAC-T) induced by hydrogen peroxide (H2O2). In vitro cell culture method was used in this experiment. Based on the oxidative stress model of MAC-T cells established by different concentrations of H2O2, the cell viability, and the level of reactive oxygen (ROS), antioxidant indexes and the mRNA expression levels of genes related to inflammation in cells were measured after treatment with different concentrations (0.15, 0.30, 0.60, 1.20, 2.40 and 4.80 mg/mL) of SBP, a control group and a H2O2 model group were set at the same time. The results showed that compared with the control group, the intracellular ROS level was extremely significantly increased after 600 μmol/L H2O2 treatment for 12 h (P < 0.01), and the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and total antioxidant capacity (T-AOC) were extremely significantly decreased (P < 0.01), while the content of MDA was extremely significantly increased (P < 0.01). After 0.15, 0.30 and 0.60 mg/mL SBP pretreatment on cells, compared with H2O2 model group, intracellular ROS level was extremely significantly decreased (P < 0.01), the T-SOD, GSH-Px and CAT activities were extremely significantly increased (P < 0.01), the T-AOC were significantly or extremely significantly increased (P < 0.05 or P < 0.01), the MDA content was extremely significantly reduced (P < 0.01), and the mRNA relative expression levels of intracellular inflammatory mediators [cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and monocyte chemotactic protein-1 (MCP-1)] and pro-inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-1β (IL-1β)] were extremely significantly decreased (P < 0.01). In conclusion, in vitro, SBP relieves oxidative stress and inflammatory damage of MAC-T cells induced by H2O2 may through reducing intracellular ROS level, improving the activities of antioxidant enzymes, and regulating the expression of inflammation-related genes.

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