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乌骨鸡(家鸡)中寡肽和四种特征肽的体外免疫调节和抗氧化作用

In vitro immunomodulatory and antioxidant effects of oligopeptides and four characteristic peptides in black-bone silky fowl (Gallus gallus domesticus Brisson).

作者信息

Zhang Zhuoran, Li Guoming, Wei Ying, Feng Zhiyuan, Fang Lei, Li Mingliang, Ren Jie, Liu Wenying, Gan Jing

机构信息

College of Life Science, Yantai University, Yantai, China.

Beijing Engineering Research Center of Protein & Functional Peptides, China National Research Institute of Food and Fermentation Industries, Beijing, China.

出版信息

J Food Biochem. 2022 Dec;46(12):e14469. doi: 10.1111/jfbc.14469. Epub 2022 Oct 7.

Abstract

Black-bone silky fowl (Gallus gallus domesticus Brisson) is considered to have strengthening effect on the body and immunomodulatory effects. The black-bone silky fowl peptide (BSFP) was produced by enzymatic digestion of the whole black-bone silky fowl (including the head and claws) after removal of the viscera. Afterwards, the four of the characteristic peptides Glu-Phe (EF), Glu-Glu-Leu (EEL), Glu-His-Pro-Thr (EHPT), Ala-Gly-Gly-His (AGGF) of the BSFP were identified by HPLC-MS/MS. The preventive effects of BSFP and the four characteristic peptides on antioxidant and immunomodulation were investigated. The antioxidant capacity was assessed by in vitro HepG2 intracellular reactive oxygen species (ROS). The immunomodulatory experiments were conducted by measuring the effects of the BSFP and four peptides on the proliferation of splenocytes, T and B lymphocytes cells, the CD4 /CD8 T lymphocytes ratio, and the phagocytic capacity of macrophages and the nitric oxide (NO) content of macrophages. The four peptides of BSFP showed strong antioxidant capacity, with the most potent peptide for intracellular ROS being AGGF, with 56% inhibition. AGGF, EF, and BSFP showed highly positive effects on splenocyte proliferation and when Concanavalin A (ConA) was used as a stimulus for T lymphocytes and lipopolysaccharide (LPS) as a stimulus for B lymphocytes, the peptides stimulated cell proliferation in a dose-dependent manner. Of these, EF, AGGF, and BSFP promoted the proliferation of T lymphocytes; EF, EHPT, and BSFP significantly promoted the proliferation of B lymphocytes. EHPT and BSFP increased the CD4 /CD8 ratio of T cells. Needle aspiration of neutral red was significantly promoted by macrophages treated with peptides other than EF. In addition, EEL, EHPT, AGGF, and BSFP had a promotive effect on NO production in phagocytes. The results indicate that BSFP is a peptide product with good immunomodulatory functions, four peptides identified from BSFP show outstanding effects in terms of antioxidant properties and immunomodulation. PRACTICAL APPLICATIONS: In this study, the amino acid composition and relative molecular masses of the black-bone silky fowl peptide were analyzed, while the four peptides with significant effects on antioxidant and immunomodulatory properties in black-bone silky fowl peptide were identified by HPLC-MS/MS technique. Positive effects of black-bone silky fowl peptide and its four peptides on antioxidant capacity and immunomodulatory ability as revealed by cell experiments. The results of this experiment provide a preliminary theoretical basis for the development of new functional foods using black-bone silky fowl peptide and their characteristic peptides.

摘要

乌鸡(家鸡)被认为具有强身健体和免疫调节作用。乌鸡肽(BSFP)是通过对去除内脏后的整只乌鸡(包括头和爪)进行酶解制备而成。之后,通过高效液相色谱-串联质谱法(HPLC-MS/MS)鉴定出了BSFP的四种特征肽,即Glu-Phe(EF)、Glu-Glu-Leu(EEL)、Glu-His-Pro-Thr(EHPT)、Ala-Gly-Gly-His(AGGF)。研究了BSFP及其四种特征肽在抗氧化和免疫调节方面的预防作用。通过体外HepG2细胞内活性氧(ROS)评估抗氧化能力。通过测量BSFP和四种肽对脾细胞、T和B淋巴细胞增殖、CD4/CD8 T淋巴细胞比值、巨噬细胞吞噬能力以及巨噬细胞一氧化氮(NO)含量的影响来进行免疫调节实验。BSFP的四种肽表现出较强的抗氧化能力,对细胞内ROS抑制作用最强的肽是AGGF,抑制率为56%。AGGF、EF和BSFP对脾细胞增殖具有高度积极的影响,当使用刀豆蛋白A(ConA)作为T淋巴细胞的刺激物以及脂多糖(LPS)作为B淋巴细胞的刺激物时,这些肽以剂量依赖的方式刺激细胞增殖。其中,EF、AGGF和BSFP促进T淋巴细胞增殖;EF、EHPT和BSFP显著促进B淋巴细胞增殖。EHPT和BSFP提高了T细胞的CD4/CD8比值。除EF外,用其他肽处理的巨噬细胞对中性红的吞噬作用明显增强。此外,EEL、EHPT、AGGF和BSFP对吞噬细胞中NO的产生具有促进作用。结果表明,BSFP是一种具有良好免疫调节功能的肽产品,从BSFP中鉴定出的四种肽在抗氧化性能和免疫调节方面表现出突出的效果。实际应用:本研究分析了乌鸡肽的氨基酸组成和相对分子质量,同时通过HPLC-MS/MS技术鉴定了乌鸡肽中对抗氧化和免疫调节性能有显著影响的四种肽。细胞实验揭示了乌鸡肽及其四种肽在抗氧化能力和免疫调节能力方面的积极作用。本实验结果为利用乌鸡肽及其特征肽开发新型功能性食品提供了初步的理论依据。

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