麦胚源活性肽对H2O2诱导的成骨细胞-破骨细胞共育体系中细胞氧化损伤的保护作用

doi:10.7506/spkx1002-6630-20211028-306

食品科学 ›› 2022, Vol. 43 ›› Issue (21): 126-136.doi: 10.7506/spkx1002-6630-20211028-306

• 营养卫生 • 上一篇

麦胚源活性肽对H2O2诱导的成骨细胞-破骨细胞共育体系中细胞氧化损伤的保护作用

罗涛，李宇，汪芳，翁泽斌，熊玲，宋海昭，王銮凤，沈新春

（1.南京财经大学食品科学与工程学院，江苏省现代粮食流通与安全协同创新中心，江苏高校粮油质量安全控制及深加工重点实验室，江苏南京 210023；2.南京中医药大学中医学院?中西医结合学院，江苏南京 210000）

发布日期:2022-12-12
基金资助:
国家自然科学基金面上项目（32172203）；江苏高校优势学科建设工程资助项目（PAPD）；江苏省研究生科研与实践创新计划项目（KYCX20_1334）

Protective Effect of Wheat Germ-Derived Peptide on H2O2-Induced Cellular Oxidative Damage in Osteoblast-Osteoclast Co-culture System

LUO Tao, LI Yu, WANG Fang, WENG Zebin, XIONG Ling, SONG Haizhao, WANG Luanfeng, SHEN Xinchun

(1. Collaborative Innovation Center for Modern Grain Circulation and Safety, Key Laboratory of Grains and Oils Quality Control and Processing, College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210023, China; 2. School of Chinese Medicine, School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210000, China)

Published:2022-12-12

摘要/Abstract

摘要： 目的：为促进麦胚的高值化利用，建立H2O2诱导的成骨细胞（osteoblast，OB）-破骨细胞（osteoclast，OC）共育体系氧化应激模型，探究麦胚源活性肽ADWGGPLPH对OB和OC活性的影响。方法：利用流式细胞术、噻唑蓝（3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide，MTT）增殖实验、异硫氰酸荧光素（fluorescein isothiocyanate，FITC）/碘化丙啶（propidium iodide，PI）双染等方法，明确麦胚源活性肽对氧化应激环境中OB增殖和凋亡的作用。利用碱性磷酸酶（alkaline phosphatase，ALP）活力检测、酶联免疫吸附测定以及抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase，TRAP）染色等方法研究麦胚源活性肽对氧化应激共育体系中OB和OC分化活性的影响。结果：麦胚源活性肽能够有效抑制氧化应激共育体系中OB内活性氧的增加，并通过抑制丙二醛生成，增强谷胱甘肽过氧化物酶、超氧化物歧化酶活力，提高OB免受自由基攻击以及清除自由基的能力。麦胚源活性肽可显著抑制氧化应激共育体系中OB凋亡（FITC/PI双染结果显示OB凋亡率由12.4%下调至5.3%，OB细胞活力由60.4%提高至92.8%（P＜0.01）），改善了OB增殖水平。此外，麦胚源活性肽对OB早期分化活性指标ALP活力、蛋白I型胶原以及晚期分化活性指标骨钙素水平的下降具有良好改善作用，OB矿化率从21.3%提高至84.3%（P＜0.01），从而使OB发挥良好的分化活性和矿化功能。TRAP染色结果显示麦胚源活性肽也能有效抑制氧化应激造成的OC过度分化（OC相对阳性面积从376.4%减小至128.1%（P＜0.01））。结论：麦胚源活性肽对H2O2诱导的OB-OC共育体系中细胞氧化损伤具有保护作用，从而维持良好的细胞稳态，本实验可为麦胚蛋白的开发利用提供一定的理论参考。

关键词: 麦胚肽；细胞氧化损伤；成骨-破骨细胞共育体系；细胞稳态

Abstract: For high-value utilization of wheat germ, the effects of wheat germ-derived peptide ADWGGPLPH on the viability of osteoblast (OB) and osteoclast (OC) in an H2O2-induced oxidative stress model were investigated in OB-OC co-culture system. Flow cytometry, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) double staining were used to clarify the effects of wheat germ-derived peptide on OB proliferation and apoptosis in oxidative stress environment. The effects of wheat germ-derived peptide on the differentiation of OB and OC in the co-culture system under oxidative stress were determined by using alkaline phosphatase (ALP) activity assay, enzyme-linked immunosorbent assay (ELISA) and tartrate resistant acid phosphatase (TRAP) staining. The results showed that wheat germ-derived peptide effectively prevented the increase of reactive oxygen species (ROS) in OB, and enhanced the ability of OB to protect against and scavenge free radicals by suppressing malondialdehyde (MDA) generation and increasing the activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). Wheat germ-derived peptide significantly inhibited the apoptosis of OB and improved its proliferation. Annexin V-FITC/PI double staining showed that the apoptosis rate was decreased from 12.4% to 5.3%, and the viability was increased from 60.4% to 92.8% (P < 0.01). In addition, wheat germ-derived peptide prevented the decrease of ALP activity, protein type I collagen (COL-I) and osteocalcin (OCN) expression, and increased OB mineralization from 21.3% to 84.3% (P < 0.01), thus maintaining OB differentiation and ensuring normal bone mineralization. Moreover, TRAP staining results showed that wheat germ-derived peptide effectively inhibited the over-differentiation of OC under oxidative stress condition, decreasing the relative positive area of OC from 376.4% to 128.1% (P < 0.01). In conclusion, this wheat germ-derived peptide has protective effects against H2O2-induced cellular oxidative damage in OB-OC co-culture system, thereby maintaining cellular homeostasis. This finding provides a certain theoretical basis for the development and utilization of wheat germ protein.

Key words: wheat germ-derived peptide; cellular oxidative damage; osteoblast-osteoclast co-culture system; cellular homeostasis

中图分类号:

TS201.2

罗涛，李宇，汪芳，翁泽斌，熊玲，宋海昭，王銮凤，沈新春. 麦胚源活性肽对H2O2诱导的成骨细胞-破骨细胞共育体系中细胞氧化损伤的保护作用[J]. 食品科学, 2022, 43(21): 126-136.

LUO Tao, LI Yu, WANG Fang, WENG Zebin, XIONG Ling, SONG Haizhao, WANG Luanfeng, SHEN Xinchun. Protective Effect of Wheat Germ-Derived Peptide on H2O2-Induced Cellular Oxidative Damage in Osteoblast-Osteoclast Co-culture System[J]. FOOD SCIENCE, 2022, 43(21): 126-136.

[1]	李祥雨熊雅婷滕建文韦保耀黄丽夏宁. 基于气相色谱-质谱技术与多元统计分析对咸蛋黄热加工中异味组分分析[J]. 食品科学, 0, (): 0-0.
[2]	钟玉旺徐万莉范尧珠黎依艳王雪峰. 辣木籽ACE抑制肽的分离纯化、结构鉴定及其体外活性评价[J]. 食品科学, 0, (): 0-0.
[3]	张俊林陈祉晴郑钦生肖杰曹庸刘晓娟. 基于分子对接和表面等离子共振技术研究虾青素与跨膜转运蛋白CD36的结合作用[J]. 食品科学, 0, (): 0-0.
[4]	王玲玉闫春梅贾梦王安琪汪轩羽王新涛李秀荷王慧芳戴福宏李宁周中凯高铁成. 油莎豆膳食纤维对短链脂肪酸生产能力和肠道菌群分布特征的影响[J]. 食品科学, 0, (): 0-0.
[5]	吴旋李娜徐怀德李梅. 晚期糖基化终末产物形成及抑制机理的研究进展[J]. 食品科学, 0, (): 0-0.
[6]	李思涵王君巧聂少平. 沙苑子多糖的提取纯化与结构解析[J]. 食品科学, 0, (): 0-0.
[7]	曹云刚，梁光灿，张鑫，范鑫，刘苗苗，冯莉，黄峻榕，熊幼翎. 碱性氨基酸调控肌原纤维蛋白加工性能及肉品品质研究进展[J]. 食品科学, 0, (): 341-348.
[8]	黄金萍，吴继红，廖小军，劳菲. 果蔬汁饮料中花色苷与VC相互作用研究进展[J]. 食品科学, 0, (): 358-371.
[9]	杨婷婷郝宗围任李顺陈光未黄艾祥. 玉米醇溶蛋白负载植物甾醇纳米颗粒的制备及性能[J]. 食品科学, 0, (): 0-0.
[10]	钟泽梁洪碧红肖美添白锴凯. 多糖纳米硒的制备、功能和在食品领域应用的研究进展[J]. 食品科学, 0, (): 0-0.
[11]	刘芙蓉王雨生李鹏房子蔚陈海华. 热诱导温度与pH值对乳清浓缩蛋白凝胶结构和性质的影响[J]. 食品科学, 0, (): 0-0.
[12]	周梦君，周舒，黄小林，李响敏，熊勇华. 酰肼介导的抗体定向偶联策略提高免疫层析试纸条检测性能[J]. 食品科学, 2022, 43(20): 44-52.
[13]	刘芙蓉，王雨生，李鹏，房子蔚，陈海华. 热诱导温度与pH值对乳清浓缩蛋白凝胶结构和性质的影响[J]. 食品科学, 2022, 43(20): 125-134.
[14]	葛艳争，石爱民，任广跃，刘哲，李嗣生，职兰懿，王强，杜祖波. 超声预处理对花生分离蛋白微凝胶颗粒结构及其Pickering乳液特性的影响[J]. 食品科学, 2022, 43(20): 95-101.
[15]	周梦君周舒黄小林熊勇华李响敏. 酰肼介导的抗体定向偶联策略提高免疫层析试纸条检测性能[J]. , 0, (): 0-0.

麦胚源活性肽对H2O2诱导的成骨细胞-破骨细胞共育体系中细胞氧化损伤的保护作用

Protective Effect of Wheat Germ-Derived Peptide on H2O2-Induced Cellular Oxidative Damage in Osteoblast-Osteoclast Co-culture System

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价