咖啡酸增强巨噬细胞免疫抑制的机制研究

doi:10.16431/j.cnki.1671-7236.2025.04.036

摘要/Abstract

摘要： 【目的】研究咖啡酸(caffeic acid，CA)对巨噬细胞免疫抑制的增强效果和调节机理，以期为研发防治免疫抑制性疾病的新型免疫增强剂提供理论支持。【方法】采用透射电子显微镜技术、纳米颗粒跟踪分析技术(nanoparticle tracking analysis，NTA)确定巨噬细胞源性外泌体(exosomes，Exos)的形态与浓度。采用磷酰胺氮芥(phosphoramide mustard，PM)建立免疫抑制模型，给予RAW264.7细胞6.25、12.5、25、50、100和200 μmol/L CA作用24 h后，通过细胞毒性试验、中性红考察细胞活力和吞噬能力、NTA测定Exos浓度、转录组学测定Exos中mRNA基因表达、GO功能与KEGG通路分析mRNA差异基因的富集情况，探讨CA增强免疫功能的机制。【结果】Exos呈完整的杯状形态，具有双层膜结构，粒径在30～200 nm之间。与PM对照组相比，CA能极显著提高巨噬细胞的活力和吞噬能力(P＜0.01)，促进Exos分泌(P＜0.01)，并上调Exos内mRNA差异基因1 961个、下调3 274个。GO功能富集分析显示，CA主要影响细胞进程、生物调节及分子结合，其功能在于正向调节树突状细胞的趋化性；KEGG通路富集分析显示，CA对免疫系统、内分泌系统、感染性疾病及信号转导具有重要作用，主要调控T细胞受体、B细胞受体相关信号通路，表明CA能通过介导Exos内mRAN表达对免疫细胞产生重要影响。【结论】CA通过促进巨噬细胞源性Exos生成，调控mRNA基因的表达，从而提升巨噬细胞的活力和吞噬能力。

关键词: 咖啡酸; 巨噬细胞; 外泌体; mRNA; 免疫抑制

Abstract: 【Objective】 The enhancement function and regulatory mechanism of caffeic acid (CA) on immunosuppression of macrophages were studied,in order to provide theoretical support for the development of a new immunoenhancer for immunosuppressive diseases (ISD).【Method】 Transmission electron microscopy and nanoparticle tracking analysis (NTA) were used to determine the morphology and concentration of macrophage-derived exosomes (Exos).Phosphoramide mustard (PM) was used to establish the immunosuppression model,RAW264.7 cells were treated with CA of 6.25,12.5,25,50, 100 and 200 μmol/L for 24 h, respectively,cytotoxicity test and neutral red were used to investigate cell viability and phagocytosis,Exos concentration was determined by NTA,mRNA expression in Exos was tested by transcriptomics,GO function and KEGG pathway were used to analyze the enrichment of mRNA differentially expressed genes,respectively,in order to explore the mechanism of CA enhancing immune function.【Result】 Exos was a complete cup shape with a bilayer membrane structure and a particle size between 30 and 200 nm. Compared with PM model group,CA could prominently enhance the activity and phagocytosis of macrophages (P＜0.01),encourage the secretion of Exos (P＜0.01),and upregulate 1 961 mRNA differentially expressed genes and downregulate 3 274 mRNA differentially expressed genes in Exos.GO function enrichment analysis showed that CA mainly affected cell process,biological regulation and molecular binding,and its function was to positively regulate the chemotaxis of dendritic cells.KEGG pathway enrichment analysis demonstrated that CA played an important role in the immune system,endocrine system,infectious diseases and signal transduction,mainly regulating T cell receptor and B cell receptor related signal pathways,indicating that CA could exert an important influence on immune cells through mediating Exos mRAN expression.【Conclusion】 CA enhanced the viability and phagocytosis of macrophages by promoting the generation of macrophage derived Exos and regulating mRNA gene expression.

Key words: caffeic acid; macrophage; exosomes; mRNA; immunosuppression

中图分类号:

白莉霞, 李洋, 李剑勇. 咖啡酸增强巨噬细胞免疫抑制的机制研究[J]. 中国畜牧兽医, 2025, 52(4): 1844-1851.

BAI Lixia, LI Yang, LI Jianyong. Study on the Mechanism of Caffeic Acid Enhancing Macrophage Immunosuppression[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(4): 1844-1851.

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