He Hong, Wang Jinghui, Cai Zhihong, Wang Mei, Wu Di, Wang Xinhui, Wang Jinqiu, He Yi, Wang Wei, Harlina Putri Widyanti, Gao Sihai, Geng Fang
Institute for Egg Science and Technology, School of Food and Biological Engineering, Chengdu University, Chengdu 610106, China.
Gastroenterology and Urology Department Ⅱ, Hunan Cancer Hospital/the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University; Clinical Research Center for Gastrointestinal Cancer in Hunan Province, Changsha 410013, China.
Poult Sci. 2025 Aug 16;104(11):105699. doi: 10.1016/j.psj.2025.105699.
Egg white glycopeptides (EWG) are dietary peptides with immune regulatory activities; however, the underlying immune regulation mechanisms of EWG-mediated have not been fully elucidated. Phosphorylation represents a crucial protein modification in immune cell signaling. In this study, phosphoproteomic technology was employed to investigate alterations in macrophage phosphoprotein profiles in response to EWG stimulation (EWG group) using lipopolysaccharide stimulation as a positive control (LPS group), and bioinformatics approaches were used to analyze the associated signaling pathways. The untreated macrophages served as the control group (CK group). Compared with the CK group, 625 differentially phosphorylated proteins (DPPs) were identified in the EWG group, involving 774 differential phosphorylation sites. Among them, the number of up-regulated DPPs was less than that of down-regulated DPPs, whereas the LPS/CK group displayed the opposite trend. This finding indicated that the mechanisms underlying EWG- and LPS-activated macrophage immunity were related to protein dephosphorylation, but the regulatory patterns of protein phosphorylation differed between EWG and LPS. In the EWG/CK group, 60.75% of DPPs were located in the nucleus and annotated to immune signaling pathways such as leukocyte transendothelial migration and Fc gamma R-mediated phagocytosis. Similar to LPS, the TLR4/MyD88/NF-κB pathway was identified to be crucial in the immune regulation of EWG-activated macrophages. This study elucidated the changes in the phosphorylated protein profiles underlying the EWG-mediated activation of macrophage immunity, providing a theoretical basis for developing EWG as an immunomodulatory food bioactive compound.
蛋清糖肽（EWG）是具有免疫调节活性的膳食肽；然而，EWG介导的潜在免疫调节机制尚未完全阐明。磷酸化是免疫细胞信号传导中一种关键的蛋白质修饰。在本研究中，采用磷酸化蛋白质组学技术，以脂多糖刺激作为阳性对照（LPS组），研究EWG刺激下巨噬细胞磷酸化蛋白质谱的变化（EWG组），并采用生物信息学方法分析相关信号通路。未处理的巨噬细胞作为对照组（CK组）。与CK组相比，EWG组鉴定出625个差异磷酸化蛋白（DPP），涉及774个差异磷酸化位点。其中，上调的DPP数量少于下调的DPP数量，而LPS/CK组则呈现相反的趋势。这一发现表明，EWG和LPS激活巨噬细胞免疫的机制与蛋白质去磷酸化有关，但EWG和LPS之间蛋白质磷酸化的调节模式不同。在EWG/CK组中，60.75%的DPP位于细胞核中，并注释到白细胞跨内皮迁移和FcγR介导的吞噬作用等免疫信号通路中。与LPS类似，TLR4/MyD88/NF-κB通路被确定在EWG激活的巨噬细胞免疫调节中起关键作用。本研究阐明了EWG介导的巨噬细胞免疫激活过程中磷酸化蛋白质谱的变化，为将EWG开发为免疫调节食品生物活性化合物提供了理论依据。
