食品科学2025，Vol.46Issue(22)：157-170,14.DOI:10.7506/spkx1002-6630-20250509-038
苦荞类外泌体纳米囊泡调控巨噬细胞RAW264.7炎症反应及极化的体外机制
Mechanism by Which Tartary Buckwheat-Derived Exosome-like Nanovesicles Regulate the Inflammatory Response and Polarization of RAW264.7 Cells in Vitro
摘要
Abstract
Objective:To investigate the regulatory effect and underlying mechanism of tartary buckwheat-derived exosome-like nanovesicles(TELN)on lipopolysaccharide(LPS)-induced inflammatory response and polarization in murine macrophage RAW264.7 cells.Methods:TELN were extracted from tartary buckwheat using size-exclusion chromatography and characterized.RAW264.7 macrophages were divided into a control,an LPS-treated group,and an LPS+TELN-treated group.Transcriptomic analysis was performed to identify the differences in gene expression between these groups.The secretion levels of the key inflammatory cytokines interleukin(IL)-6,tumor necrosis factor-α(TNF-α),IL-1β,and IL-10 in cell culture supernatants were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of these cytokines was detected by real-time quantitative polymerase chain reaction(qPCR).The impact of TELN on LPS-induced macrophage polarization was analyzed using flow cytometry and Western blot.Results:TELN exhibited a homogeneous size distribution with an average diameter of 145.8 nm and a concentration of(1.305±0.074)×1010 particles/mL.The transcriptomic analysis revealed significant upregulation of the genes encoding macrophage-derived chemotactic factors,ubiquitinated proteins,phosphorylated proteins,and immune/inflammatory regulators(e.g.,S100A8)in the TELN-treated group.The ELISA results demonstrated that compared with the LPS group,TELN significantly reduced the levels of the pro-inflammatory cytokines IL-6,TNF-α,and IL-1β in supernatants by(75.6±0.9)%(P<0.000 1),(10.9±0.2)%(P<0.000 1),and(57.8±6.8)%(P<0.000 1),respectively,while elevating the anti-inflammatory cytokine IL-10 by(69.7±4.6)%(P<0.000 1).The qPCR results confirmed that TELN significantly downregulated the mRNA expression of IL-6,TNF-α,and IL-1β and upregulated IL-10 mRNA expression at the transcriptional level.Flow cytometry and Western blot analyses consistently showed that TELN significantly suppressed the LPS-induced expression of the M1 polarization marker CD86.However,no detectable CD206 protein expression was observed,indicating that TELN did not promote M2 polarization.Conclusion:TELN exerts an anti-inflammatory effect by bidirectionally modulating cytokine secretion(effectively inhibiting the release of the pro-inflammatory cytokines IL-6,TNF-α,and IL-1β while significantly enhancing the level of anti-inflammatory IL-10)and suppressing M1 macrophage polarization.Particularly,TELN has a pronounced inhibitory effect on IL-6 expression,suggesting that its mechanism of action may be related to targeting signal pathways such as nuclear factor kappa-B(NF-κB).This study provides novel insights into the nutritional and immunomodulatory mechanisms of tartary buckwheat.关键词
苦荞来源类外泌体纳米囊泡/巨噬细胞RAW264.7/转录组学/生物信息学/炎症Key words
tartary buckwheat-derived exosome-like nanovesicles/RAW264.7 macrophages/transcriptomics/bioinformatics/inflammation分类
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曹亚楠,刘乙志,彭镰心..苦荞类外泌体纳米囊泡调控巨噬细胞RAW264.7炎症反应及极化的体外机制[J].食品科学,2025,46(22):157-170,14.基金项目
四川省科技厅计划项目(2023ZYD0078) （2023ZYD0078）
现代农业产业技术体系四川杂粮创新团队专项(SCCXTD-2024-20) （SCCXTD-2024-20）
西南民族大学青藏高原民族药用资源保护与利用实验室开放基金项目(QTPEM2201) （QTPEM2201）
