Page 7 - 南京医科大学学报自然科学版

第43卷第11期 南京医科大学学报（自然科学版）
2023年11月 Journal of Nanjing Medical University（Natural Sciences） ·1479 ·

·基础研究·

羟氯喹介导NF⁃κB/NLRP3通路抑制M1型巨噬细胞极化缓解小

鼠肠道炎症

王 舒 ，江婧月 ，王 迪 ，林俊杰 ，赵小静 ，张红杰 1*
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南京医科大学第一附属医院消化内科，江苏 南京 210029；徐州医科大学附属医院消化内科，江苏 徐州 221000
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［摘 要］ 目的：探讨羟氯喹（hydroxychloroquine，HCQ）对葡聚糖硫酸钠（dextran sulfate sodium salt，DSS）诱导的结肠炎模型鼠
肠道炎症的影响及其 M1 型巨噬细胞极化机制。方法：C57BL/6 小鼠随机分 3 组：对照组（正常饮水+200 μL 纯水每日灌胃）、
DSS组（自由饮用3.5% DSS溶液+200 μL纯水每日灌胃）、DSS+HCQ组［自由饮用3.5% DSS+200 μL HCQ溶液（60 mg/kg）每日
灌胃］。造模期间观察小鼠粪便性状，记录体重、疾病活动指数（disease activity index，DAI）评分。造模后测量小鼠结肠长度、
结肠组织HE染色行组织病理学评分；提取结肠组织固有层单个核细胞，流式细胞术检测M1型巨噬细胞比例。体外提取并诱
导分化小鼠骨髓来源的巨噬细胞，HCQ 处理后，流式细胞术检测 M1 型巨噬细胞比例；蛋白质印迹法检测 p⁃STAT1、IRF5、
NF⁃κB/p65等表达水平。结果：HCQ可以减轻DSS诱导的小鼠肠道炎症并减少结肠中M1型巨噬细胞比例。细胞实验显示HCQ可
抑制M1型巨噬细胞极化并同时抑制NF⁃κB信号通路及其下游NOD样受体热蛋白结构域相关蛋白3（NOD⁃like receptor thermal
protein domain associated protein 3，NLRP3）炎症小体的形成；NF⁃κB通路抑制剂PDTC可得到相似的结果。结论：羟氯喹可抑制
NF⁃κB/NLRP3信号通路减少M1型巨噬细胞极化，减轻DSS诱导的小鼠肠道炎症。
［关键词］ 溃疡性结肠炎；羟氯喹；巨噬细胞极化
［中图分类号］ R574.62 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）11⁃1479⁃08
doi：10.7655/NYDXBNS20231101

Hydroxychloroquine ameliorates colitis in mice by inhibiting M1 macrophage polarization

through NF⁃κB/NLRP3 signaling pathway
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WANG Shu ，JIANG Jingyue ，WANG Di ，LIN Junjie ，ZHAO Xiaojing ，ZHANG Hongjie 1*
1 Department of Gastroenterology，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029；
Department of Gastroenterology，the Affiliated Hospital of Xuzhou Medical University，Xuzhou 221000，China
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［Abstract］ Objective：The current study aims to explore the effect of hydroxychloroquine（HCQ）on dextran sulfate sodium salt（DSS）⁃
induced colitis in mice and to reveal the underlying mechanisms of HCQ in the inhibition of M1 macrophage polarisation. Methods：All
the C57BL/6 mice were randomly divided into 3 groups：the control group（common drinking water+200 μL pure water by gavage），DSS
group（3.5% DSS drinking water+200 μL pure water by gavage）and DSS+HCQ group（3.5% DSS drinking water+200 μL HCQ of 60
mg/kg by gavage）. The disease activity index（DAI）score was evaluated according to stool property and body weight changes every
day. The mice in each group were sacrificed after modeling experiment with the colon length measured. The colon tissues were stained
with haematoxylin and eosin（HE）for histological score calculation. The colonic lamina propria mononuclear cells were isolated and
the proportion of M1 macrophages was measured by flow cytometry. In vitro，bone marrow⁃derived macrophages were isolated and
treated with HCQ，and the proportion of M1 macrophages was measured by flow cytometry. The levels of p⁃STAT1，IRF5，NF⁃κB /p65，
etc in the macrophages were detected by Western blot. Results：HCQ ameliorated DSS⁃induced colitis and significantly decreased the
percentage of M1 macrophages in colon in DSS ⁃ induced colitis mice. In vitro，HCQ significantly repressed M1 polarisation and
decreased the phosphorylation of NF ⁃ κB and the downstream of NOD ⁃ like receptor thermal protein domain associated protein 3
（NLRP3）inflammasome formation in M1 macrophages. The similar results were observed with PDTC（an inhibitor of NF⁃κB signaling）
treatment. Conclusion：HCQ attenuates colonic inflammation in mice by supressing M1 macrophage polarization through the inhibition
［基金项目］ 国家自然科学基金（82070568，82200582）
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通信作者（Corresponding author），E⁃mail：hjzhang06@163.com