食品科学 ›› 2018, Vol. 39 ›› Issue (13): 153-158.doi: 10.7506/spkx1002-6630-201813023

• 营养卫生 • 上一篇    下一篇

蛋清源活性肽抗氧化及抗炎活性

张 燕,魏汝君,潘风光,刘 珍,王寒松,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:2018-07-15 发布日期:2018-07-09
  • 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)

Antioxidant and Anti-Inflammatory Effects of Bioactive Peptides Derived from Egg White Proteins

ZHANG Yan, WEI Rujun, PAN Fengguang, LIU Zhen, WANG Hansong, LIU Jingbo*   

  1. College of Food Science and Engineering, Jilin University, Changchun 130062, China
  • Online:2018-07-15 Published:2018-07-09

摘要: 食源性抗氧化肽安全性高,抗氧化能力较好,近年来成为多肽研究领域的热点。本实验以蛋清源抗 氧化肽WNWAD(Trp-Asn-Trp-Ala-Asp),及其拆分肽WNW(Trp-Asn-Trp)、WAD(Trp-Ala-Asp)、WN (Trp-Asn)、AD(Ala-Asp)为研究对象,首先采用2’-联氨-双-3-乙基苯并噻唑啉-6-磺酸(2,2’-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid),ABTS)体外抗氧化评价方法,对5 条肽的抗氧化活性进行了评价,建立H2O2 诱导的人胚肾细胞(HEK293)氧化损伤模型,利用HEK293细胞模型评价5 条肽的抗氧化活性,然后检测抗氧化 肽WNWAD对细胞氧化型谷胱甘肽(oxidized glutathione,GSSG)含量的影响,最后利用酶联免疫吸附测定试剂盒 检测了炎症因子人白细胞介素-8及人肿瘤坏死因子的含量。结果表明:活性肽浓度为40 μmol/L时,WNWAD清除 ABTS+?能力最强,Trolox抗氧化当量(Trolox equivalent antioxidant capacity,TEAC)为(98.411±0.020)μmol TE/L, 其次为WN、WNW、WAD,其TEAC分别为(96.629±0.008)、(94.978±0.003)、(88.807±0.003)μmol TE/L, AD没有ABTS+?清除能力。在H2O2浓度为400 μmol/L时,细胞相对存活率为(50.240±0.505)%,有高度显著性差 异(P<0.001),不同浓度的蛋清源活性肽对H2O2诱导的HEK293细胞氧化损伤均有保护作用,且随着各条肽浓度 升高保护作用呈增强趋势,其中WNWAD的保护作用最强,浓度为30 μmol/L时,细胞存活率已高达99.12%。并且 WNWAD可明显降低细胞中GSSG含量,降低炎症因子人白细胞介素-8和人肿瘤坏死因子的含量。以上研究结果显 示蛋清源活性肽对降低HEK293细胞氧化应激水平、提高细胞抗氧化能力、抗炎能力等有重要作用。

关键词: 蛋清源活性肽, 2’-联氨-双-3-乙基苯并噻唑啉-6-磺酸, HEK293细胞, 抑制氧化损伤, 谷胱甘肽, 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 antioxidant and anti-inflammatory effects of a bioactive peptide derived from egg white proteins, WNWAD (Trp-Asn-Trp-Ala-Asp), and its separations WNW (Trp-Asn-Trp), WAD (Trp-Ala-Asp), WN (Trp-Asn) and AD (Ala-Asp). The protective effect against oxidative stress induced by H2O2 in HEK293 cells was evaluated and the in vitro antioxidant activity was measured by 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. The impact of WNWAD on the cellular content of oxidized glutathione (GSSG) was tested and its effect on the levels of interleukin-8 (IL-8) and tumor necrosis factor (TNF) was assessed by using commercial enzyme-linked immunosorbent assay kits. The results showed that WNWAD at 40 μmol/L possessed the strongest ABTS radical scavenging capacity with a Trolox equivalent antioxidant capacity (TEAC) value of (98.411 ± 0.020)μmol TE/L, followed by WN, WNW and WAD, whose TEAC values were (96.629 ± 0.008), (94.978 ± 0.003) and (88.807 ± 0.003) μmol TE/L, respectively, while AD had no ABTS radical scavenging potential. The survival rate of the cells was (50.240 ± 0.505)% at a H2O2 concentration of 400 μmol/L, with a statistically significant difference (P < 0.001). All peptides concentration dependently protected HEK293 cells from H2O2-induced oxidative damage. Among these, WNWAD had the strongest cytoprotective activity and the cell survival rate was up to 99.12% at 30 μmol/L. Moreover, WNWAD pronouncedly decrease the cellular GSSG content and lowered the levels of IL-8 and TNF. The results above demonstrated that WNWAD played an important role in reducing oxidative stress in HEK293 cells, enhancing cellular antioxidant activity and resisting inflammations.

Key words: egg white bioactive peptide, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), HEK293 cell, protection against oxidative damage, glutathione, cytokine

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