摘要
目的:分析比较鳀鱼来源和大豆来源蛋白肽产品的多肽组成及其对小鼠肠黏膜免疫功能的调节活性。方法:采用超高效液相色谱-高分辨多级质谱联用技术,结合Peaks Online 1.7软件,解析鳀鱼蛋白肽和大豆蛋白肽的所有多肽序列,并比较其多肽组成的差异。采用环磷酰胺诱导肠黏膜免疫抑制小鼠模型,分析比较鳀鱼蛋白肽和大豆蛋白肽对小鼠小肠黏膜结构、小肠黏膜免疫因子、免疫球蛋白水平和Toll样受体(Toll-like receptors,TLRs)/核因子-κB(nuclear factor kappa B,NF-κB)信号通路相关因子表达量的影响。结果:鳀鱼蛋白肽和大豆蛋白肽的多肽组成有很大差异,只有11%的多肽序列相同。与模型组相比,补充鳀鱼蛋白肽和大豆蛋白肽(300 mg/kg mb)30 d可以显著提高小鼠脾脏指数和胸腺指数,减轻环磷酰胺造成的肠黏膜结构损伤,增加小肠绒毛长度,减小隐窝深度。同时,补充鳀鱼蛋白肽和大豆蛋白肽还能够改善环磷酰胺对小肠黏膜中免疫因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、干扰素-γ(interferon-gamma,IFN-γ)、白细胞介素-4(interleukin-4,IL-4)和IL-13表达的抑制,上调免疫球蛋白A和分泌型免疫球蛋白A的分泌,并上调小肠中TLR-4、TLR-6、TLR-9和NF-κB基因的表达。结论:鳀鱼蛋白肽和大豆蛋白肽可能是通过保护TLRs/NF-κB信号通路,进而促进小肠黏膜中免疫因子和免疫球蛋白的分泌,改善环磷酰胺对肠黏膜结构的损伤和肠黏膜免疫功能的抑制。虽然鳀鱼蛋白肽和大豆蛋白肽的重均分子质量、氨基酸组成以及多肽组成有较大差异,但除了鳀鱼蛋白肽对肠黏膜分泌型免疫球蛋白A的保护效果优于大豆蛋白肽外,鳀鱼蛋白肽和大豆蛋白肽在保护肠黏膜结构和调节小鼠肠黏膜免疫方面的活性无明显差异。
Objective:This study aimed to compare the compositions of peptides from Engraulis japonicus and Glycine max and their immunomodulatory effects on the intestinal mucosa in mice.Methods:The sequences of peptides from E.japonicus and G.max were analyzed by ultra-performance liquid chromatography-high resolution mass spectrometry(UPLC-MS/MS)combined with Peaks Online 1.7 software,and the differences in peptide composition were compared.The effects of the two peptides on the mucosal structure,immune factors,immunoglobulin(Ig)levels,and gene expression of factors associated with the Toll-like receptors(TLRs)/nuclear factor kappa B(NF-κB)signaling pathway in the small intestine of mice with cyclophosphamide-induced immunosuppression were investigated.Results:The peptide compositions from E.japonicus and G.max were quite different.Only 11%of the peptide sequences were identical between them.Administration of the peptides from E.japonicus and G.max at 300 mg/kg mb for 30 d could significantly increase the spleen index and thymus index,alleviate intestinal mucosal damage,increase the length of intestinal villi and decrease the depth of crypts in cyclophosphamide-induced immunosuppressed mice.Moreover,the inhibitory effect of cyclophosphamide on the expression of tumor necrosis factor-α(TNF-α),interferon-gamma(IFN-γ),interleukin-4(IL-4),and IL-13 was ameliorated,and the secretion of IgA and secretory IgA and the gene expression of TLR-4,TLR-6,TLR-9 and NF-κB were upregulated in the small intestine mucosa.Conclusion:The peptides from E.japonicus and G.max can promote the secretion of immune factors and immunoglobulins in the small intestinal mucosa and alleviate cyclophosphamide-induced intestinal mucosal damage and immunosuppression in mice by protecting the TLRs/NF-κB signaling pathway.Although the average molecular masses,amino acid compositions and polypeptide compositions of the two peptides are different and the protective effect of the peptides from E.japonicu on secretory IgA in the intestinal mucosa is more pronounced than that of the peptides from G.max,there is no significant difference between their efficacy in protecting intestinal mucosal structure and regulating mucosal immunity in mice.
作者
刘银媛
王寒梅
逯良忠
沈金华
杨颖
赵雪
LIU Yinyuan;WANG Hanmei;LU Liangzhong;SHEN Jinhua;YANG Ying;ZHAO Xue(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China;Zhonghai Ocean Science&Technology Co.,Ltd.,Qingdao 266000,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2023年第19期107-117,共11页
Food Science
基金
国家自然科学基金面上项目(32172137)
青岛中海海洋生物资源开发有限公司技术开发项目(20210248)
山东省泰山学者青年专家计划项目(tsqn202103033)。
关键词
鳀鱼
大豆
蛋白肽
肽组成
肠黏膜免疫
免疫球蛋白
Engraulis japonicas
Glycine max
peptide
peptide composition
intestinal mucosal immunity
immunoglobulin
作者简介
第一作者:刘银媛(1998-)(ORCID:0000-0002-7308-0270),女,硕士研究生,研究方向为海洋活性物质。E-mail:L15531266322@126.com;通信作者:赵雪(1976-)(ORCID:0000-0003-1855-2543),女,副教授,博士,研究方向为海洋活性物质。E-mail:zhaoxue@ouc.edu.cn。
