人参来源细胞外囊泡通过抑制cGAS-STING通路调控辐射诱导的巨噬细胞极化与炎症反应

doi:10.7506/spkx1002-6630-20260302-015

摘要/Abstract

摘要： 目的：探讨人参来源细胞外囊泡（ginseng-derived extracellular vesicles，Gin-DEVs）对辐射诱导的RAW264.7巨噬细胞极化与炎症反应的影响及其潜在作用机制。方法：采用超速离心结合蔗糖密度梯度法从鲜人参中提取并纯化Gin-DEVs，通过透射电镜与纳米粒径仪表征其形态、粒径与电位。采用CCK-8法检测不同质量浓度Gin-DEVs对RAW264.7巨噬细胞活力的影响并筛选最适给药浓度；通过荧光示踪法观察RAW264.7细胞对Gin- DEVs的摄取情况。此外，建立6 Gy γ射线辐射损伤模型，将RAW264.7巨噬细胞分为空白对照组、模型组及Gin- DEVs干预组，利用流式细胞术检测M1型（CD80＋CD206－）与M2型（CD80＋CD206＋）巨噬细胞比例；通过酶联免疫吸附测定（enzyme-linked immunosorbent assay，ELISA）法测定细胞上清液中肿瘤坏死因子-α（tumor necrosis factor-alpha，TNF‑α）、白细胞介素（interleukin，IL）-1β、IL-10及一氧化氮（nitric oxide，NO）水平，通过2’,7’-二氯二氢荧光素二乙酸酯（2’,7’-dichlorodihydrofluorescein diacetate，DCFH-DA）荧光探针法检测细胞内活性氧（reactive oxygen species，ROS）水平；通过Western blot法分析干扰素基因刺激因子（stimulator of interferon genes，STING）及其下游因子TANK结合激酶1（TANK-binding kinase 1，TBK1）、磷酸化TBK1（phosphorylated- TBK1，p-TBK1）、干扰素调节因子3（interferon regulatory factor 3，IRF3）、磷酸化IRF3（phosphorylated-IRF3，p-IRF3）的表达。结果：纯化后的Gin-DEVs呈典型双层膜结构，粒径集中在（155.67±0.38）nm，颗粒浓度为1.0×10 12 Particles/mL。CCK8分析结果显示，Gin-DEVs对RAW264.7细胞的增殖具有促进作用，且具有浓度依赖效应。荧光示踪实验证实Gin-DEVs可被RAW264.7细胞有效摄取。流式细胞术分析结果显示，Gin-DEVs可显著抑制辐射诱导的M1型巨噬细胞极化，促进M2型极化（P＜0.05）。ELISA分析结果显示，与模型组相比，Gin-DEVs可显著降低TNF-α、IL-1β及NO水平，提高IL-10水平（P＜0.01）。DCFH-DA荧光探针分析结果显示，Gin-DEVs可显著抑制辐射诱导的细胞内ROS积累（P＜0.01）。Western blot分析结果显示，与模型组比较，Gin-DEVs可显著抑制STING、p-TBK1、p-IRF3蛋白的表达（P＜0.05）。结论：Gin-DEVs可能通过抑制环磷酸鸟苷-腺苷合成酶-STING信号通路的激活，调节辐射诱导的巨噬细胞极化，抑制炎症反应与氧化应激。

关键词: 人参来源细胞外囊泡；辐射损伤；巨噬细胞极化；炎症反应；环磷酸鸟苷-腺苷合成酶-干扰素刺激基因通路

Abstract: Objective: To investigate the effects of ginseng-derived extracellular vesicles (Gin-DEVs) on radiation-induced polarization and inflammatory responses in RAW264.7 macrophages and to explore the underlying mechanisms. Methods: Gin-DEVs were extracted and purified from fresh ginseng using ultracentrifugation combined with sucrose density gradient centrifugation. Their morphology, particle size, and zeta potential were characterized by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The effects of different concentrations of Gin-DEVs on the viability of RAW264.7 macrophages were assessed using the CCK-8 assay to determine the optimal concentration for subsequent experiments. The uptake of Gin-DEVs by RAW264.7 cells was observed using fluorescence tracing. A radiation injury model was established by exposing cells to 6 Gy γ-ray irradiation. RAW264.7 cells were divided into a blank control group, a model group, and a Gin-DEVs intervention group. The proportions of M1-type (CD80 +CD206–) and M2-type (CD80+CD206+) macrophages were determined by flow cytometry. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-10, and nitric oxide (NO) in the cell culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Intracellular reactive oxygen species (ROS) levels were detected using a fluorescent probe, 2’,7’- dichlorodihydrofluorescein diacetate (DCFH-DA). The protein expression levels of stimulator of interferon genes (STING) and its downstream factors, TANK-binding kinase 1 (TBK1), phosphorylated TBK1 (p-TBK1), interferon regulatory factor 3 (IRF3), and phosphorylated IRF3 (p-IRF3), were analyzed by Western blot. Results: The purified Gin-DEVs exhibited a typical bilayer membrane structure, with an average particle size of (155.67 ± 0.38) nm and a particle concentration of 1.0 × 10 12 particles/mL. CCK-8 results indicated that Gin-DEVs promoted the proliferation of RAW264.7 cells in a concentration-dependent manner. Flow cytometry results showed that Gin-DEVs significantly inhibited radiation-induced M1 macrophage polarization and promoted M2 polarization (P < 0.05). ELISA results demonstrated that compared with the control group, Gin-DEVs significantly reduced the levels of TNF-α, IL-1β, and NO but increased the level of IL-10 (P < 0.01). DCFH-DA fluorescent probe results revealed that Gin-DEVs significantly suppressed radiation-induced intracellular ROS accumulation (P < 0.01). Western blot analysis revealed that compared with the model group, Gin-DEVs significantly suppressed the protein expression of STING, p-TBK1, and p-IRF3 (P < 0.05). Conclusion: Gin-DEVs may regulate radiation-induced macrophage polarization and suppress inflammatory responses and oxidative stress by inhibiting the activation of the cyclic GMP-AMP synthase (cGAS)-STING signaling pathway.

Key words: ginseng-derived extracellular vesicles; radiation damage; macrophage polarization; inflammatory response; cyclic GMP-AMP synthase-stimulator of interferon genes pathway

中图分类号:

R285.5

陈祉君，秦兰，于兴泰，关晓娴，王文艺，杨锦璇，耿淑慧，范玲帆，翟永松，郑宇，陆洋. 人参来源细胞外囊泡通过抑制cGAS-STING通路调控辐射诱导的巨噬细胞极化与炎症反应[J]. 食品科学, 2026, 47(19): 213-221 https://doi.org/10.7506/spkx1002-6630-20260302-015

CHEN Zhijun, QIN Lan, YU Xingtai, GUAN Xiaoxian, WANG Wenyi, YANG Jinxuan, GENG Shuhui, FAN Lingfan, ZHAI Yongsong, ZHENG Yu, LU Yang. Ginseng-Derived Extracellular Vesicles Regulate Radiation-Induced Macrophage Polarization and Inflammatory Responses through Suppressing the cGAS-STING Signaling Pathway[J]. FOOD SCIENCE, 2026, 47(19): 213-221 https://doi.org/10.7506/spkx1002-6630-20260302-015

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人参来源细胞外囊泡通过抑制cGAS-STING通路调控辐射诱导的巨噬细胞极化与炎症反应

Ginseng-Derived Extracellular Vesicles Regulate Radiation-Induced Macrophage Polarization and Inflammatory Responses through Suppressing the cGAS-STING Signaling Pathway

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