食品科学 ›› 2023, Vol. 44 ›› Issue (23): 142-150.doi: 10.7506/spkx1002-6630-20221110-115

• 营养卫生 • 上一篇    下一篇

牦牛乳酪蛋白抗氧化肽对HEK293细胞氧化应激损伤的保护作用

吴登宇, 韦体, 马忠仁, 宋礼, 杨具田, 蔡勇, 高丹丹   

  1. (1.西北民族大学生物医学研究中心,中国-马来西亚国家联合实验室,甘肃 兰州 730030;2. 西北民族大学生命科学与工程学院,甘肃 兰州 730124;3.甘肃华羚乳品股份有限公司,甘肃 合作 747000)
  • 出版日期:2023-12-15 发布日期:2024-01-02
  • 基金资助:
    西北民族大学中央高校基本科研业务费资金资助项目(31920230153);甘肃省高校青年博士支持项目(2023QB-001); 兰州市城关区科技计划项目(2022JSCX0011);甘肃省科技厅项目科技专员专项(23CXGA0078); 兰州市人才创新创业项目(2023-QN-69);甘肃省科技厅-科技型中小企业创新基金(23CXGP0002)

Protective Effect of Antioxidant Peptides Derived from Yak Milk Casein on Oxidative Stress Injury in HEK293 Cells

WU Dengyu, WEI Ti, MA Zhongren, SONG Li, YANG Jutian, CAI Yong, GAO Dandan   

  1. (1. China-Malaysia National Joint Laboratory, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China; 2. College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730124, China; 3. Gansu Hualing Dairy Co., Ltd., Hezuo 747000, China)
  • Online:2023-12-15 Published:2024-01-02

摘要: 本实验采用H2O2诱导HEK293细胞建立细胞氧化损伤模型,确定H2O2最佳处理浓度和时间,研究5 条牦牛乳酪蛋白抗氧化肽(AFK、IEQI、FPFF、LPVPQ、RELEEL)对H2O2诱导损伤HEK293细胞存活率、丙二醛含量、抗氧化酶类活力、还原型谷胱甘肽和氧化型谷胱甘肽含量的影响,并探讨牦牛乳酪蛋白抗氧化肽的体外抗氧化作用机制,为其在高附加值生物制品及功能食品领域的开发与应用提供理论依据。结果表明:牦牛乳酪蛋白抗氧化肽对不同的自由基具有不同的清除作用,但均呈剂量效应关系;使用终浓度为400 μmol/L的H2O2处理HEK293细胞12 h后,对细胞的抑制率为(46.21±0.40)%;细胞毒性实验分析结果表明,5 条抗氧化肽对HEK293细胞既无毒副作用,也无促进增殖作用;牦牛乳酪蛋白抗氧化肽能够显著降低HEK-293氧化损伤细胞中的丙二醛(除LPVPQ)、氧化型谷胱甘肽含量,并增强抗氧化酶活性,其中200 μg/mL抗氧化肽RELEEL能够显著降低丙二醛含量至(0.062±0.000)nmol/104 cells,提升谷胱甘肽含量至(61.17±2.48)μg/106 cells,并维持较高的谷胱甘肽/氧化型谷胱甘肽值(64.93±0.95);200 μg/mL抗氧化肽LPVPQ能够显著降低氧化型谷胱甘肽含量至(0.74±0.26)μg/106 cells,增强超氧化物歧化酶活力至(1.17±0.02)U/104 cells;200 μg/mL抗氧化肽AFK能够显著增强过氧化氢酶活力至(0.60±0.09)U/104 cells。上述结果显示,牦牛乳酪蛋白抗氧化肽对氧化损伤的细胞具有积极影响,可为下一步相关产品的开发提供参考。

关键词: 牦牛乳酪蛋白;抗氧化肽;HEK293细胞;氧化损伤;保护作用

Abstract: In this experiment, HEK293 cells were induced by H2O2 to establish a cell model of oxidative damage, and the optimal H2O2 concentration and treatment time were determined. The effects of five antioxidant peptides (AFK, IEQI, FPFF, LPVPQ and RELEEL) derived from yak milk casein on the survival rate, malondialdehyde (MDA) content, antioxidant enzyme activities, reduced glutathione and oxidized glutathione contents of the damaged cells were studied, and the mechanism of action of the antioxidant peptides was explored to provide a theoretical basis for their development and application in high value-added biological products and functional foods. The results showed that these antioxidant peptides had different scavenging effects on different radical species, but they all showed a dose-effect relationship. After being treated with a final concentration of 400 μmol/L H2O2 for 12 h, the inhibitory rate of HEK293 cells was (46.21 ± 0.40)%. Cytotoxicity test showed that the five antioxidant peptides had no toxic or side effects on HEK-293 cells, and did not promote cell proliferation either. These antioxidant peptides could significantly reduce the contents of malondialdehyde (except LPVPQ) and oxidized glutathione in HEK293 cells damaged by oxidation, and enhance the activity of antioxidant enzymes. RELEEL at a concentration of 200 μg/mL significantly reduced MDA content to (0.062 ± 0.000) nmol/104 cells and increased glutathione content to (61.17 ± 2.48) μg/106 cells while maintaining high GSH/GSSG ratio of 64.93 ± 0.95. The antioxidant peptide LPVPQ at 200 μg/mL significantly reduced the content of oxidized glutathione to (0.74 ± 0.26) μg/106 cells and increased the activity of superoxide dismutase (SOD) to (1.17 ± 0.02) U/104 cells; the antioxidant peptide AFK at 200 μg/mL significantly enhanced catalase (CAT) activity to (0.60 ± 0.09) U/104 cells. These results indicate that the antioxidant peptides derived from yak milk casein have positive effects on cells damaged by oxidation, which can provide a reference for further development of relevant products.

Key words: yak milk casein; antioxidant peptides; HEK293 cells; oxidative damage; protective effect

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