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

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

本试验旨在探究大豆活性肽(SBP)对体外过氧化氢(H_(2)O_(2))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H_(2)O_(2)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H_(2)O_(2)模型组。结果显示:与对照组相比,600μmol/L的H_(2)O_(2)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H_(2)O_(2)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H_(2)O_(2)诱导的MAC-T细胞的氧化应激和炎症损伤。

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(H_(2)O_(2)).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 H_(2)O_(2),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 H_(2)O_(2) 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 600μmol/L H_(2)O_(2) 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 H_(2)O_(2) 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 H_(2)O_(2) may through reducing intracellular ROS level,improving the activities of antioxidant enzymes,and regulating the expression of inflammation-related genes.