基于巨噬细胞炎症模型探讨榆干离褶伞溶栓酶的抗炎作用

doi:10.7506/spkx1002-6630-20250520-128

食品科学 ›› 2025, Vol. 46 ›› Issue (20): 199-207.doi: 10.7506/spkx1002-6630-20250520-128

基于巨噬细胞炎症模型探讨榆干离褶伞溶栓酶的抗炎作用

苏新，张晴，王佳怡，沈明花

（1.延边大学医学院，吉林延吉 133002；2.通化医药健康职业学院，吉林通化 134001）

出版日期:2025-10-25 发布日期:2025-09-17
基金资助:
吉林省自然科学基金资助项目（YDZJ202201ZYTS212）

Anti-inflammatory Effect of Lyophyllum ulmarium Fibrinolytic Enzyme on Macrophages

SU Xin, ZHANG Qing, WANG Jiayi, SHEN Minghua

(1. Medical College, Yanbian University, Yanji 133002, China; 2. Tonghua Medical and Health Vocational College, Tonghua 134001, China)

Online:2025-10-25 Published:2025-09-17

摘要/Abstract

摘要： 基于脂多糖（lipopolysaccharide，LPS）诱导的J774A.1巨噬细胞炎症模型，探究榆干离褶伞溶栓酶（Lyophyllum ulmarium fibrinolytic enzyme，LUFE）的抗炎作用。将J774A.1细胞分为正常对照组、LPS组以及LPS＋LUFE低和高剂量组（LPS＋LUFE-L组、LPS＋LUFE-H组）。LPS＋LUFE-L组和LPS＋LUFE-H组细胞先以10、20 μg/mL的LUFE培养24 h后，除正常对照组以外的其余各组均以1 μg/mL的LPS处理24 h。酶联免疫吸附法检测细胞白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor alpha，TNF-α）以及细胞培养上清液的单核细胞趋化蛋白-1 （monocyte chemotactic protein-1，MCP-1）水平；细胞迁移实验观察细胞趋化能力；细胞与细胞外基质实验检测细胞黏附功能；中性红吞噬实验检测细胞吞噬能力；免疫荧光染色观察细胞活性氧（reactive oxygen species，ROS）水平；比色法检测细胞丙二醛（malondialdehyde，MDA）含量、过氧化氢酶（catalase，CAT）及超氧化物歧化酶（superoxide dismutase，SOD）的活性。蛋白免疫印迹方法测定Toll样受体4（toll-like receptor 4，TLR4）及核因子相关因子2（nuclear factor erythroid 2-related factor，Nrf2）通路相关蛋白的表达情况。结果表明，LUFE干预后能够降低IL-6、IL-1β、MCP-1、ROS、MDA水平，抑制LPS所致巨噬细胞的趋化、黏附及吞噬功能的提高，增强细胞SOD、CAT活性。同时，下调TLR4、髓样分化因子88（myeloid differentiation primary response protein 88，MyD88）、Kelch样ECH关联蛋白1及核因子κB蛋白表达及活化水平；上调Nrf2、血红素加氧酶1、NADPH醌氧化还原酶1水平。综上，LUFE可通过调控Nrf2及TLR4/MyD88信号通路抑制脂多糖诱导的巨噬细胞炎症反应。

关键词: 榆干离褶伞；溶栓酶；炎症；氧化应激；巨噬细胞

Abstract: This study evaluated the anti-inflammatory effect of a fibrinolytic enzyme from Lyophyllum ulmarium (LUFE) on lipopolysaccharide-induced J774A.1 macrophages. The cells were divided into four groups: normal control, LPS, LPS + low-dose LUFE (LUFE-L), and LPS + high-dose (LUFE-H). The LPS + LUFE-L and LPS + LUFE-H groups were first incubated with 10 and 20 μg/mL of LUFE for 24 h, respectively, and then all groups except the normal control group were treated with 1 μg/mL of LPS for 24 h. Enzyme-linked immunosorbent assay (ELISA) was conducted to determine interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and monocyte chemotactic protein-1 (MCP-1) levels in cell culture supernatants; the Transwell assay was used to observe cell chemotaxis ability; cell adhesion function was detected by the cell-extracellular matrix assay and cell phagocytosis by neutral red phagocytosis assay; immunofluorescence staining was performed to observe the level of cellular reactive oxygen species (ROS); colorimetric assays were used to detect cellular malondialdehyde (MDA) and catalase (CAT) levels and superoxide dismutase (SOD) activity. Western blotting was performed to determine the expression of Toll-like receptor 4 (TLR4) and nuclear factor erythroid 2-related factor (Nrf2) pathway-related proteins. The results showed that LUFE intervention reduced the levels of IL-6, IL-1β, MCP-1, ROS, and MDA, inhibited LPS-induced macrophage chemotaxis, adhesion, and phagocytosis, and elevated cellular SOD and CAT levels. Meanwhile, it down-regulated the expression and activation of TLR4, myeloid differentiation primary response protein 88 (MyD88), Kelch-like ECH-associated protein 1 (Keap-1) and nuclear factor kappa-B (NF-κB) proteins and up-regulated the levels of Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H-quinone oxidoreductase 1 (NQO1). In summary, LUFE could inhibit LPS-induced inflammatory responses in macrophages by modulating the Nrf2 and TLR4/MyD88 signaling pathways.

Key words: Lyophyllum ulmarium; thrombolytic enzyme; inflammation; oxidative stress; macrophage

中图分类号:

R282.71

苏新，张晴，王佳怡，沈明花. 基于巨噬细胞炎症模型探讨榆干离褶伞溶栓酶的抗炎作用[J]. 食品科学, 2025, 46(20): 199-207.

SU Xin, ZHANG Qing, WANG Jiayi, SHEN Minghua. Anti-inflammatory Effect of Lyophyllum ulmarium Fibrinolytic Enzyme on Macrophages[J]. FOOD SCIENCE, 2025, 46(20): 199-207.

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基于巨噬细胞炎症模型探讨榆干离褶伞溶栓酶的抗炎作用

Anti-inflammatory Effect of Lyophyllum ulmarium Fibrinolytic Enzyme on Macrophages

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