表没食子儿茶素没食子酸酯通过NF-κB通路抑制脂多糖诱导的巨噬细胞向M1表型极化

doi:10.7506/spkx1002-6630-201801022

食品科学 ›› 2018, Vol. 39 ›› Issue (1): 142-148.doi: 10.7506/spkx1002-6630-201801022

表没食子儿茶素没食子酸酯通过NF-κB通路抑制脂多糖诱导的巨噬细胞向M1表型极化

吴琼1，王乐锋1，张妍淞2，汤小芳2，张贤益2，李露2，舒瑶2，黄成2，廖金珠2，王富龙2，李文娟2,*

1.南昌大学第二附属医院，江西南昌 330006；2.南昌大学食品科学与技术国家重点实验室，江西南昌 330047

出版日期:2018-01-15 发布日期:2018-01-05
基金资助:
国家自然科学基金地区科学基金项目（31560460）；江西省科技厅自然科学基金项目（20151512041185）； “三区”人才支持计划科技特派人员专项（0210208651）

EGCG Inhibits LPS-Caused Polarization of Macrophages into M1 Phenotype via the NF-κB Pathway

WU Qiong1, WANG Lefeng1, ZHANG Yansong2, TANG Xiaofang2, ZHANG Xianyi2, LI Lu2, SHU Yao2, HUANG Cheng2, LIAO Jinzhu2, WANG Fulong2, LI Wenjuan2,*

1. The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China; 2. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China

Online:2018-01-15 Published:2018-01-05

摘要/Abstract

摘要： 目的：研究表没食子儿茶素没食子酸酯（(-)-epigallocatechin gallate，EGCG）的抗炎作用及其对巨噬细胞极化的影响。方法：原代培养小鼠腹腔巨噬细胞，通过对细胞培养基进行不同的处理，将实验分为对照组、脂多糖（lipopolysaccharides，LPS，1 μg/mL）组、EGCG（25 μmol/L）组和EGCG＋LPS（25 μmol/L＋1 μg/mL）组；流式细胞仪检测吞噬活性、活性氧（reactive oxygen species，ROS）含量和细胞凋亡情况，噻唑蓝法检测细胞增殖情况；Griess法测定NO的含量；酶联免疫吸附法测定肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和白细胞介素-1β（interleukin-1β，IL-1β）含量，Western blot检测Toll-样受体4（Toll like receptor 4，TLR4）和核转录因子-κB（nuclear transcription factor-κB，NF-κB）p65蛋白表达量。结果：EGCG和/或LPS对小鼠腹腔巨噬细胞增殖和凋亡无显著影响；与LPS组相比，EGCG可以极显著抑制LPS刺激腹腔巨噬细胞中ROS、NO、TNF-α、IL-1β增加（P＜0.01），提示EGCG可抑制巨噬细胞向M1表型极化，具有抗炎作用；与LPS组相比，EGCG＋LPS组中巨噬细胞TLR4和细胞核蛋白中NF-κB p65表达量明显下降（P＜0.01）。结论：EGCG对LPS致炎的小鼠腹腔巨噬细胞具有抗炎作用，其作用机制可能与其阻断TLR4受体介导的NF-κB通路活化，抑制巨噬细胞向M1表型极化有关。

关键词: 表没食子儿茶素没食子酸酯, 抗炎作用, 细胞表型, NF-κB通路, 巨噬细胞极化

Abstract: Objective: The aim of this study was to explore the anti-inflammatory effect of (-)-epigallocatechin gallate (EGCG) and its effects on macrophage polarization. Methods: Primary cultures of mouse peritoneal macrophages were divided into 4 groups: control, 1 μg/mL lipopolysaccharides (LPS), 25 μmol/L EGCG, and 25 μmol/L EGCG + 1 μg/mL LPS groups. Flow cytometry analysis was used to determine phagocytosis, reactive oxygen species (ROS) and apoptosis. Cell proliferation was monitored by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. NO generation was analyzed by Griess method. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme linked immunosorbent assay (ELISA). The expression of Toll-like receptor 4 (TLR4) and NF-κB p65 proteins was detected by using Western blot analysis. Results: EGCG and/or LPS were no significant effect on cell proliferation or apoptosis in mouse peritoneal macrophages. Compared with the LPS group, EGCG could significantly inhibit the LPS-induced increasing in ROS, NO, TNF-α and IL-1β in peritoneal macrophages (P < 0.01), thereby consequently suppress the polarization of macrophages into M1 phenotype and consequently exerting anti-inflammatory effects. Compared with the LPS group, the expression of TLR4 protein in macrophages and the expression of nuclear transcription factor-κB (NF-κB) p65 in nucleus were significantly decreased in the EGCG + LPS group (P < 0.01). Conclusion: EGCG could inhibit LPS-induced inflammatory responses in mouse peritoneal macrophages, and the underlying mechanism may be related to the blocking of the polarization of macrophages into M1 phenotype via the TLR4-mediated NF-κB pathway.

Key words: (-)-epigallocatechin gallate, anti-inflammatory effects, cell phenotype, NF-κB pathway, macrophage polarization

中图分类号:

TS201.4

吴琼，王乐锋，张妍淞，汤小芳，张贤益，李露，舒瑶，黄成，廖金珠，王富龙，李文娟. 表没食子儿茶素没食子酸酯通过NF-κB通路抑制脂多糖诱导的巨噬细胞向M1表型极化[J]. 食品科学, 2018, 39(1): 142-148.

WU Qiong, WANG Lefeng, ZHANG Yansong, TANG Xiaofang, ZHANG Xianyi, LI Lu, SHU Yao, HUANG Cheng, LIAO Jinzhu, WANG Fulong, LI Wenjuan. EGCG Inhibits LPS-Caused Polarization of Macrophages into M1 Phenotype via the NF-κB Pathway[J]. FOOD SCIENCE, 2018, 39(1): 142-148.

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表没食子儿茶素没食子酸酯通过NF-κB通路抑制脂多糖诱导的巨噬细胞向M1表型极化

EGCG Inhibits LPS-Caused Polarization of Macrophages into M1 Phenotype via the NF-κB Pathway

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