食品科学 ›› 2019, Vol. 40 ›› Issue (5): 130-136.doi: 10.7506/spkx1002-6630-20171103-035

• 营养卫生 • 上一篇    下一篇

蛋清源活性肽对过氧化氢诱导的HEK293细胞抗氧化酶活力及白细胞介素8分泌的影响

张 燕,胡 榕,郑 健,马中苏,杨太芬,刘静波*   

  1. 吉林大学食品科学与工程学院,吉林 长春 130062
  • 出版日期:2019-03-15 发布日期:2019-04-02
  • 基金资助:
    国家自然科学基金面上项目(31271907;31471597);高等学校博士学科点专项科研基金(博导类)项目(20130061110088)

Effects of Egg White-Derived Bioactive Peptide on Antioxidative Enzyme Activities and IL-8 Secretion in HEK293 Cells under Oxidative Stress Induced by Hydrogen Peroxide

ZHANG Yan, HU Rong, ZHENG Jian, MA Zhongsu, YANG Taifen, LIU Jingbo*   

  1. College of Food Science and Engineering, Jilin University, Changchun 130062, China
  • Online:2019-03-15 Published:2019-04-02

摘要: 食源性抗氧化肽由于抗氧化能力较好、安全性高,逐渐成为多肽研究领域的热点。本实验以H2O2诱导HEK293细胞损伤建立氧化损伤模型,研究了蛋清源活性肽WNWAD(Trp-Asn-Trp-Ala-Asp)及其结构改造肽WNW(Trp-Asn-Trp)、WAD(Trp-Ala-Asp)、WN(Trp-Asn)对H2O2诱导损伤的HEK293细胞的存活率、活性氧(reactive oxygen species,ROS)水平、抗氧化酶活力及细胞因子白介素8(interleukin-8,IL-8)分泌的影响。结果表明:蛋清源活性肽对HEK293细胞存活率的影响极显著(P<0.01),不同浓度蛋清源活性肽WNWAD、WNW、WAD、WN作用于氧化损伤的HEK293细胞后,其存活率明显提高,呈现浓度依赖性;其中1.0 μmol/L蛋清源活性肽WNWAD、WNW、WAD、WN处理细胞后,存活率从损伤组的(48.0±2.4)%分别提高到(99.7±1.8)%、(69.4±3.2)%、(78.9±5.1)%、(72.1±3.6)%,与损伤组有极显著性差异(P<0.01);H2O2诱导HEK293细胞内ROS的产生,经过活性肽预处理后细胞内ROS的水平极显著降低(P<0.01);随着蛋清源活性肽WNWAD、WNW、WAD、WN浓度的升高,总超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶3 种酶的活力均有所提高,呈现浓度依赖型关系。最后利用酶联免疫吸附测定试剂盒测定了HEK293细胞中IL-8的分泌量,经过H2O2处理的HEK293细胞上清液中IL-8含量极显著升高(P<0.01),加入WNWAD、WNW、WN、WAD后,细胞中的IL-8含量均有所降低,其中WNWAD和WN影响显著(P<0.05)。以上结果显示蛋清源活性肽对降低HEK293细胞氧化应激水平和提高细胞抗氧化能力有积极的影响。

关键词: 蛋清源活性肽, HEK293细胞, 活性氧含量, 抗氧化酶系, 细胞因子

Abstract: In recent years, antioxidant peptides with high safety and potent antioxidant capacity have become a hot research field. The aim of this study was to investigate the effects of egg white-derived bioactive peptides WNWAD (Trp-Asn-Trp- Ala-Asp), WNW (Trp-Asn-Trp), WAD (Trp-Ala-Asp) and WN (Trp-Asn) on the survival rate, reactive oxygen species (ROS) content, antioxidative enzyme activities and interleukin-8 (IL-8) level in HEK293 cells under oxidative stress induced by H2O2. MTS assay results showed that all four peptides significantly dose-dependently inhibited H2O2-induced cellular oxidative stress in HEK-293 cells and increased the survival rate increased from (48.0 ± 2.4)% to (99.7 ± 1.8)%, (69.4 ± 3.2)%, (78.9 ± 5.1)%, and (72.1 ± 3.6)% at 1.0 μmol/L concentration, respectively. These changes were statistically significant (P < 0.01). 2’,7’-Dichlorofluorescin diacetate (DCFH-DA) fluorescent probe assay proved that all these peptides inhibited oxidative stress by reducing intracellular ROS accumulation in HEK293 cells. They dose-dependently increased the activity of total superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in HEK293 cells. In addition, the results of enzyme-linked immune sorbent assay (ELISA) showed that WNWAD and WN up-regulated IL-8 secretion (P < 0.05). All the above results showed that the egg white-derived peptides play a positive role in protecting HEK293 cells against H2O2-induced oxidative stress and improving antioxidant capacity in HEK293 cells.

Key words: egg white-derived peptide, HEK293 cell, reactive oxygen species content, antioxidant enzyme system, cytokines

中图分类号: