The Human Milk-derived Peptide Drives Rapid Regulation of Macrophage Inflammation Responses in the Neonatal Intestine.

IF 7.1 1区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY Cellular and Molecular Gastroenterology and Hepatology Pub Date : 2024-10-15 DOI:10.1016/j.jcmgh.2024.101420

Fuqiang Yuan, Xu Han, Masha Huang, Yinglin Su, Yiting Zhang, Mengyuan Hu, Xiang Yu, Weilai Jin, Yun Li, Le Zhang

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Abstract

Background & aims: The interactions between human milk and the regulation of innate immune homeostasis in newborns, and their impact on intestinal health, are not fully understood. This study aimed to explore the role of peptides in human milk extracellular vesicles (EVs) in this process.

Methods: A comprehensive screening of peptides within human milk EVs was performed, leading to the identification of a beta-casein-derived peptide (CASB_135-150). The effects of CASB_135-150 on intestinal injury were evaluated in a rat necrotizing enterocolitis (NEC) model. Immunofluorescence analysis was used to determine its distribution, and its impact on NF-κB signaling and inflammation was studied in bone marrow-derived macrophages (BMDMs) and intestinal macrophages. Protein-protein interaction (PPI) analysis, single-cell RNA-seq (scRNA-seq), and co-immunoprecipitation (co-IP) experiments were conducted to explore the mechanism underlying CASB_135-150 function.

Results: CASB_135-150 significantly mitigated intestinal injury in the rat NEC model. Immunofluorescence analysis revealed that CASB_135-150 could target intestinal macrophages and rapidly inhibited NF-κB signaling and reduced inflammation. ScRNA-seq analyses indicated a strong association between FHL2 and NEC development, and co-IP confirmed the interaction between CASB_135-150 and FHL2. CASB_135-150 disrupted the FHL2/TRAF6 complex, reducing TRAF6 protein levels. Mutation of key amino acids in CASB_135-150 disrupted its interaction with FHL2 and abolished its ability to inhibit NF-κB signaling, which also prevented its protective effect in vivo. RNA-seq of intestinal tissue further highlighted the impact of CASB_135-150 on the NF-κB signaling pathway.

Conclusions: Our study identifies CASB_135-150, a novel peptide in human milk EVs, that rapidly regulates macrophage inflammatory responses and protects against NEC-induced intestinal injury. These findings provide new insights into the role of human milk in modulating the infant immune system and intestinal health.

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人乳衍生肽驱动新生儿肠道巨噬细胞炎症反应的快速调节

背景和目的：母乳与新生儿先天性免疫平衡调节之间的相互作用及其对肠道健康的影响尚不完全清楚。本研究旨在探索人乳细胞外囊泡 (EVs) 中的多肽在这一过程中的作用：方法：对人乳细胞外囊泡中的多肽进行了全面筛选，最终确定了一种来源于β-酪蛋白的多肽（CASB135-150）。在大鼠坏死性小肠结肠炎（NEC）模型中评估了 CASB135-150 对肠道损伤的影响。免疫荧光分析确定了 CASB135-150 的分布，并研究了它对骨髓源性巨噬细胞（BMDMs）和肠道巨噬细胞中 NF-κB 信号转导和炎症的影响。为了探索CASB135-150的功能机制，研究人员进行了蛋白-蛋白相互作用（PPI）分析、单细胞RNA-seq（scRNA-seq）和共免疫沉淀（co-IP）实验：结果：CASB135-150能明显减轻大鼠NEC模型的肠道损伤。免疫荧光分析表明，CASB135-150 可靶向肠道巨噬细胞，迅速抑制 NF-κB 信号传导并减轻炎症反应。ScRNA-seq分析表明FHL2与NEC的发生密切相关，co-IP证实了CASB135-150与FHL2之间的相互作用。CASB135-150 干扰了 FHL2/TRAF6 复合物，降低了 TRAF6 蛋白水平。CASB135-150中关键氨基酸的突变破坏了它与FHL2的相互作用，取消了它抑制NF-κB信号转导的能力，这也阻止了它在体内的保护作用。肠组织的RNA-seq进一步强调了CASB135-150对NF-κB信号通路的影响：我们的研究发现了人乳EVs中的一种新型多肽CASB135-150，它能快速调节巨噬细胞的炎症反应，并对NEC诱导的肠道损伤起到保护作用。这些发现为了解母乳在调节婴儿免疫系统和肠道健康方面的作用提供了新的视角。

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来源期刊

Cellular and Molecular Gastroenterology and Hepatology Medicine-Gastroenterology

CiteScore

13.00

自引率

2.80%

发文量

246

审稿时长

42 days

期刊介绍： "Cell and Molecular Gastroenterology and Hepatology (CMGH)" is a journal dedicated to advancing the understanding of digestive biology through impactful research that spans the spectrum of normal gastrointestinal, hepatic, and pancreatic functions, as well as their pathologies. The journal's mission is to publish high-quality, hypothesis-driven studies that offer mechanistic novelty and are methodologically robust, covering a wide range of themes in gastroenterology, hepatology, and pancreatology. CMGH reports on the latest scientific advances in cell biology, immunology, physiology, microbiology, genetics, and neurobiology related to gastrointestinal, hepatobiliary, and pancreatic health and disease. The research published in CMGH is designed to address significant questions in the field, utilizing a variety of experimental approaches, including in vitro models, patient-derived tissues or cells, and animal models. This multifaceted approach enables the journal to contribute to both fundamental discoveries and their translation into clinical applications, ultimately aiming to improve patient care and treatment outcomes in digestive health.