小麦抗氧化肽对人胚肾细胞氧化应激损伤的抑制作用及分子机制

doi:10.12141/j.issn.1000-565X.230082

华南理工大学学报(自然科学版) ›› 2024, Vol. 52 ›› Issue (4): 33-41.doi: 10.12141/j.issn.1000-565X.230082

小麦抗氧化肽对人胚肾细胞氧化应激损伤的抑制作用及分子机制

刘文颖¹ 任杰² 吴晗硕² 韩鲁佳^1†

^1.中国农业大学工学院，北京 100083
^2.中国食品发酵工业研究院有限公司北京市蛋白功能肽工程技术研究中心，北京 100015

收稿日期:2023-03-03 出版日期:2024-04-25 发布日期:2023-08-18
通信作者: 韩鲁佳（1964-），女，博士，教授，主要从事农产品加工工程研究。 E-mail:hanlj@cau.edu.cn
作者简介:刘文颖（1984-），女，博士生，高级工程师，主要从事农产品加工工程研究。E-mail:wenyingliu888@126.com
基金资助:
宁夏回族自治区重点研发计划项目(2021BEG02027)

Inhibitory Effects and Molecular Mechanism of Wheat Antioxidant Peptides on Oxidative Stress Injury in Human Embryonic Kidney Cells

LIU Wenying¹ REN Jie² WU Hanshuo² HAN Lujia¹

^1.College of Engineering，China Agricultural University，Beijing 100083，China
^2.Beijing Engineering Research Center of Protein and Functional Peptides，China National Research Institute of Food and Fermentation Industries Co. ，Ltd. ，Beijing 100015，China

Received:2023-03-03 Online:2024-04-25 Published:2023-08-18
Contact: 韩鲁佳（1964-），女，博士，教授，主要从事农产品加工工程研究。 E-mail:hanlj@cau.edu.cn
About author:刘文颖（1984-），女，博士生，高级工程师，主要从事农产品加工工程研究。E-mail:wenyingliu888@126.com
Supported by:
the Key R&D Program of Ningxia Hui Autonomous Region(2021BEG02027)

摘要/Abstract

摘要：

通过2，2-偶氮二异丁基脒二盐酸盐（AAPH）诱导建立人胚肾细胞（HEK293）氧化应激损伤模型，评价5种小麦蛋白源肽Leu-Tyr（LY）、Pro-Tyr（PY）、Tyr-Gln（YQ）、Ala-Pro-Ser-Tyr（APSY）和Arg-Gly-Gly-Tyr（RGGY）的抗氧化活性，然后利用量子化学计算和分子对接技术预测5种小麦蛋白源肽与2，2-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐（ABTS）结合的最优构型和结合作用，探讨小麦蛋白源肽发挥活性的分子机制。细胞试验结果表明：5种小麦蛋白源肽作用后，细胞死亡率显著下降到3.68%以下（P < 0.05），并且能显著减少AAPH诱导的活性氧自由基（ROS）生成（P < 0.05），使ROS含量趋于正常水平；5种小麦蛋白源肽均呈现出较好的总抗氧化能力和1，1-二苯基-2-三硝基苯肼（DPPH）自由基清除能力（P < 0.05），RGGY的总抗氧化能力最强，活性值为（1.46 ± 0.08）mmol/L Trolox，其次是APSY、YQ、PY和LY；YQ的DPPH自由基清除能力最强，清除率为61.34% ± 2.24%，其次是APSY、RGGY、PY和LY。分子对接结果表明，5种小麦蛋白源肽的CDOCKER交互能量（-CIE）评分分别为13.304 9、13.397 3、13.412 1、16.768 5、16.268 3，均可以有效地与ABTS相互作用，主要通过与ABTS分子之间形成较强的氢键和疏水作用力来发挥抗氧化活性。

关键词: 小麦蛋白源肽, 抗氧化剂, 细胞培养, 分子对接技术, 分子机制

Abstract:

The study firstly established oxidative stress injury model of human embryonic kidney cells (HEK293) by the induction of 2,2-azobis (2-methylpropionamidine) dihydrochloride (AAPH) to evaluate the antioxidant activity of five wheat protein-derived peptides Leu-Tyr (LY), Pro-Tyr (PY), Tyr-Gln (YQ), Ala-Pro-Ser-Tyr (APSY) and Arg-Gly-Gly-Tyr (RGGY). Then, it used quantum chemistry and molecular docking techniques to predict the optimal configuration and binding effect of five wheat protein-derived peptides combined with 2,2-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), and to explore the molecular mechanism of wheat protein-derived peptides. The results of cell test show that after the action of five wheat protein-derived peptides, the cell death rate significantly decreases to below 3.68% (P < 0.05), and the generation of reactive oxygen species (ROS) induced by AAPH was significantly reduced (P < 0.05), making the ROS content tend to normal levels. All five wheat protein-derived peptides show good total antioxidant capacity and free radical scavenging capacity of 1,1-Diphenyl-2-picrylhydrazyl (DPPH) (P < 0.05). RGGY shows the strongest total antioxidant capacity, with an activity value of (1.46 ± 0.08) mmol/L Trolox, followed by APSY, YQ, PY and LY. The DPPH free radical scavenging ability of YQ is the strongest, with a scavenging rate of 61.34% ± 2.24%, followed by APSY, RGGY, PY and LY. The results of molecular docking show that the CDOCKER interaction energy (-CIE) scores of the five wheat protein-derived peptides are 13.304 9, 13.397 3, 13.412 1, 16.768 5 and 16.268 3, respectively, which can effectively interact with ABTS, mainly through the formation of strong hydrogen bonds and hydrophobic forces between ABTS molecules to exert antioxidant activity.

Key words: wheat protein-derived peptide, antioxidant, cell culture, molecular docking technology, molecular mechanism

中图分类号:

TS201.4

刘文颖, 任杰, 吴晗硕, 等. 小麦抗氧化肽对人胚肾细胞氧化应激损伤的抑制作用及分子机制[J]. 华南理工大学学报(自然科学版), 2024, 52(4): 33-41.

LIU Wenying, REN Jie, WU Hanshuo, et al. Inhibitory Effects and Molecular Mechanism of Wheat Antioxidant Peptides on Oxidative Stress Injury in Human Embryonic Kidney Cells[J]. Journal of South China University of Technology(Natural Science Edition), 2024, 52(4): 33-41.

图/表 9

图1

图2

图3

图4

图5

图6

图7

图8

图9

参考文献 34

1	ZHAO X C， ZHANG L， YU H X，et al ．Curcumin protects mouse neuroblastoma Neuro-2A cells against hydrogen-peroxide-induced oxidative stress［J］．Food Chemistry，2011，129（2）：387-394.
2	LUSHCHAK V I ．Free radicals，reactive oxygen species，oxidative stress and its classification［J］．Chemico-Biological Interactions，2014，224：164-175.
3	YI G F， DIN J U， ZHAO F，et al ．Effect of soybean peptides against hydrogen peroxide induced oxidative stress in HepG2 cells via Nrf2 signaling［J］．Food & Function，2020，11（3）：2725-2737.
4	刘文颖，冯晓文，程青丽，等．大米低聚肽的制备和结构表征及体外抗氧化作用［J］．华南理工大学学报（自然科学版），2021，49（11）：47-56.
	LIU Wenying， FENG Xiaowen， CHENG Qingli，et al ．Preparation，structure characterization and in vitro antioxidant activity of rice oligopeptides［J］．Journal of South China University of Technology （Natural Science Edition），2021，49（11）：47-56.
5	NORDBERG J， ARNER E S J ．Reactive oxygen species，antioxidants，and the mammalian thioredoxin system［J］．Free Radical Biology and Medicine，2001，31（11）：1287-1312.
6	吉正梅，张晓春，彭钰迪，等．鸭胚源抗氧化肽TD12对HepG2细胞氧化应激损伤的保护作用［J］．食品与发酵工业，2021，47（18）：141-148.
	JI Zhengmei， ZHANG Xiaochun， PENG Yudi，et al ．Protective effect of duck embryo-derived antioxidant peptide TD12 on oxidative stress damage in HepG2 cells ［J］．Food and Fermentation Industries，2021，47（18）：141-148.
7	KARAMI Z， PEIGHAMBARDOUST S H， HESARI J，et al ．Antioxidant，anticancer and ACE-inhibitory activities of bioactive peptides from wheat germ protein hydrolysates［J］．Food Bioscience，2019，32：100450/1-12.
8	ZHANG J， WEN C， LI C，et al ．Antioxidant peptide fractions isolated from wheat germ protein with subcritical water extraction and its transport across Caco-2 cells［J］．Journal of Food Science，2019，84（8）：2139-2146.
9	LIU W Y， ZHANG J T， TAKUYA MIYAKAWA，et al ．Antioxidant properties and inhibition of angiotensin-converting enzyme by highly active peptides from wheat gluten［J］．Scientific Reports，2021，11（1）：5206/1-11.
10	张燕，陈志飞，赵颂宁，等．蛋清源抗氧化肽对HEK293细胞氧化应激损伤的抑制作用及机制［J］．中国食品学报，2019，19（10）：11-22.
	ZHANG Yan， CHEN Zhifei， ZHAO Songning，et al ．The anti-oxidative effects and mechanism of antioxidant peptides from egg white against oxidative stress injury in human embryonic kidney 293 cells［J］．Journal of Chinese Institute of Food Science and Technology，2019，19（10）：11-22.
11	ZHANG Q， CUI C， CHEN C Q，et al ．Anti-proliferative and proapoptotic activities of Alpinia oxyphylla on HepG2 cells through ROS-mediated signaling pathway［J］．Journal of Ethnopharmacology，2015，169：99-108.
12	LIU E Y， FANG L， FENG X W，et al ． In vitro antioxidant and angiotensin I-converting enzyme inhibitory properties of peptides derived from corn gluten meal ［J］．European Food Research and Technology，2020，246：2017-2027.
13	MINE Y， MA F P， LAURIAU S ．Antimicrobial peptides released by enzymatic hydrolysis of hen egg white lysozyme［J］．Journal of Agricultural and Food Chemistry，2004，52（5）：1088-1094.
14	WEN C， ZHANG J， ZHANG H，et al ．Study on the structure-activity relationship of watermelon seed antioxidant peptides by using molecular simulations［J］．Food Chemistry，2021，364：130432/1-7.
15	ZHANG J， LI M， ZHANG G，et al ．Identification of novel antioxidant peptides from snakehead （Channa argus） soup generated during gastrointestinal digestion and insights into the anti-oxidation mechanisms［J］．Food Chemistry，2021，337：127921/1-11.
16	LI C， ZHAN Y D， MA X Z，et al ．B7-H4 facilitates proliferation and metastasis of colorectal carcinoma cell through PI3K/Akt/mTOR signaling pathway［J］．Clinical and Experimental Medicine，2020，20（1）：79-86.
17	LIU L， QIU T X， SONG D W，et al ．Inhibition of a novel coumarin on an aquatic rhabdovirus by targeting the early stage of viral infection demonstrates potential application in aquaculture［J］．Antiviral Research，2020，174：104672/1-13.
18	LU Q B， LIN X Y， WU J，et al ．Matrine attenuates cardiomyocyte ischemia-reperfusion injury through activating AMPK/Sirt3 signaling pathway［J］．Journal of Receptors and Signal Transduction Research，2021，41（1）：488-493.
19	LU J M， LIN P H， YAO Q Z，et al ．Chemical and molecular mechanisms of antioxidants：experimental approaches and model systems［J］．Journal of Cellular and Molecular Medicine，2010，14（4）：840-860.
20	NEMOTO S， TAKEDA K， YU Z X，et al ．Role for mitochondrial oxidants as regulators of cellular metabolism［J］．Molecular and Cellular Biology，2000，20（19）：7311-7318.
21	YU D L， ZHA Y Y， ZHONG Z，et al ．Improved detection of reactive oxygen species by DCFH-DA：new insight into self-amplification of fluorescence signal by light irradiation［J］．Sensors and Actuators B：Chemical，2021，339：129878/1-9.
22	LIU X， FU Q， XU P，et al ．Rapid determination of monopersulfate with bromide ion-catalyzed oxidation of 2，2′-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid （ABTS）［J］．Chemical Engineering Journal，2022，433：133551/1-8.
23	QIN X Y， ZHANG J T， LIU G M，et al ．Structure and composition of a potential antioxidant obtained from the chelation of pea oligopeptide and sodium selenite［J］．Journal of Functional Foods，2019，64：103619/1-8.
24	CHANG O K， HA G E， HAN G S，et al ．Novel antioxidant peptide derived from the ultrafiltrate of ovomucin hydrolysate［J］．Journal of Agricultural and Food Chemistry，2013，61：7294-7300.
25	SUN J， HE H， XIE B J ．Novel antioxidant peptides from fermented mushroom Ganoderma lucidum［J］．Journal of Agricultural and Food Chemistry，2004，52：6646-6652.
26	CHEN H M， MURAMOTO K， YAMAUCHI F，et al ．Structural analysis of antioxidative peptides from soybean beta-conglycinin［J］．Journal of Agricultural and Food Chemistry，1995，43：574-578.
27	SAITO K， JIN D H， OGAWA T，et al ．Antioxidative properties of tripeptide libraries prepared by the combinatorial chemistry［J］．Journal of Agricultural and Food Chemistry，2003，51：3668-3674.
28	GU R Z， LIU W Y， LIN F，et al ．Antioxidant and angiotensin I-converting enzyme inhibitory properties of oligopeptides derived from black-bone silky fowl （Gallus gallus domesticus Brisson） muscle［J］．Food Research International，2012，49：326-333.
29	ZHOU C， HU J， MA H，et al ．Antioxidant peptides from corn gluten meal：orthogonal design evaluation ［J］．Food Chemistry，2015，187（15）：270-278.
30	ZHU K X， ZHOU H M， QIAN H F ．Antioxidant and free radical-scavenging activities of wheat germ protein hydrolysates （WGPH） prepared with alcalase［J］．Process Biochemistry，2006，41（6）：1296-1302.
31	张燕，魏汝君，潘风光，等．蛋清源活性肽抗氧化及抗炎活性［J］．食品科学，2018，39（13）：153-158.
	ZHANG Yan， WEI Rujun， PAN Fengguang，et al ．Antioxidant and anti-inflammatory effects of bioactive peptides derived from egg white proteins［J］．Food Science，2018，39（13）：153-158.
32	WU R， HUANG J， HUAN R，et al ．New insights into the structure-activity relationships of antioxidative peptide PMRGGGGYHY［J］．Food Chemistry，2021，337：127678/1-8.
33	文超婷．西瓜籽肽的抗氧化构效关系及其分子机制研究［D］．镇江：江苏大学，2021.
34	YANG Q， CAI X， YAN A，et al ．A specific antioxidant peptide：its properties in controlling oxidation and possible action mechanism［J］．Food Chemistry，2020，327：126984/1-9.

[1]	赵强忠冯梦莹林恋竹赵谋明. 牛大力糖蛋白的分离、鉴定及抗氧化活性[J]. 华南理工大学学报（自然科学版）, 2015, 43(11): 8-15.
[2]	吴振强腾惠兰 Kwong C W 吴建勇. 生物反应器内人参细胞的培养生长及代谢特性[J]. 华南理工大学学报（自然科学版）, 2006, 34(5): 38-42.

小麦抗氧化肽对人胚肾细胞氧化应激损伤的抑制作用及分子机制

Inhibitory Effects and Molecular Mechanism of Wheat Antioxidant Peptides on Oxidative Stress Injury in Human Embryonic Kidney Cells

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 34

相关文章 2

编辑推荐

Metrics

本文评价