蛇床子素对脂多糖诱导的巨噬细胞极化和炎症反应的影响

doi:10.19439/j.sjos.2022.03.006

上海口腔医学 ›› 2022, Vol. 31 ›› Issue (3): 255-259.doi: 10.19439/j.sjos.2022.03.006

蛇床子素对脂多糖诱导的巨噬细胞极化和炎症反应的影响

王菲菲¹, 饶定成², 金思宇², 杨沛鸣¹, 周永梅^3#, 万光勇^4#

1.潍坊医学院口腔医学院,山东潍坊 261053;
2.上海交通大学医学院,上海 200011;
3.上海交通大学医学院附属第九人民医院口腔黏膜病科,上海交通大学口腔医学院,国家口腔医学中心, 国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011;
4.泰安市中心医院口腔科,山东泰安 271000

收稿日期:2021-10-14 修回日期:2022-02-22 出版日期:2022-06-25 发布日期:2022-07-07
通讯作者: 周永梅,E-mail:jinyuanwang39@hotmail.com;万光勇, E-mail:wgykq@163.com。^#共同通信作者
作者简介:王菲菲（1991-）,女,在读硕士研究生,E-mail:1848308168@qq.com
基金资助:
上海市教委“地高大”计划（20152226）; 黄浦区产业扶持基金（XK2020013）; 上海市申康项目（SHDC2020CR3060B）; 上海交通大学医学院大学生科创项目（1420Y503）

The effect of osthole on lipopolysaccharide-induced macrophages polarization and inflammatory reaction

WANG Fei-fei¹, RAO Ding-cheng², JIN Si-yu², YANG Pei-ming¹, ZHOU Yong-mei³, WAN Guang-yong⁴

1. College of Stomatology, Weifang Medical University. Weifang 261053, Shandong Province;
2. Shanghai Jiao Tong University School of Medicine. Shanghai 200011;
3. Department of Oral Medicine, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Center of Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011;
4. Department of Stomatology, Tai'an Central Hospital. Tai'an 271000, Shandong Province, China

Received:2021-10-14 Revised:2022-02-22 Online:2022-06-25 Published:2022-07-07

摘要/Abstract

摘要： 目的: 探讨蛇床子素(osthole, OST)对脂多糖（lipopolysaccharide, LPS）诱导的巨噬细胞极化、炎症反应的影响。方法: 以CCK-8法检测不同浓度OST对RAW264.7巨噬细胞增殖活性的影响;使用细胞内活性氧（reactive oxygen species, ROS）检测试剂盒（DCFH-DA）、免疫荧光染色、q-PCR、流式细胞术明确不同浓度（6.25、12.5、25 μmol/L）OST对巨噬细胞极化和炎症反应的影响。采用Graphpad prism 8.0软件包对数据进行统计学分析。结果: CCK-8结果显示,在25 μmol/L以下时,OST对RAW264.7无明显细胞毒性。免疫荧光染色及q-PCR结果显示,6.25、12.5、25 μmol/L的OST均能抑制M1型巨噬细胞炎症因子的表达,iNOS、TNF-α、CCR7呈浓度依赖性降低,并且有效上调M2抑炎因子IL-10、Arg-1及CD206的表达。流式细胞仪分析表明,OST可有效抑制LPS诱导的巨噬细胞M1型标志物CD86的表达。结论: OST调节LPS诱导的M1型巨噬细胞极化,减轻炎症反应。

关键词: 蛇床子素, 炎性骨病, 巨噬细胞极化, 脂多糖

Abstract: PURPOSE: To investigate the effect of Osthole (OST) on lipopolysaccharide (LPS)-induced macrophage polarization and inflammatory reaction. METHODS: The effect of different concentrations of OST on proliferative activity of RAW264.7 macrophages was examined by CCK-8 method; the effect of OST at different concentrations (6.25, 12.5 and 25 μmol/L) on macrophage polarization and inflammation was investigated by using intracellular reactive oxygen species (ROS) detection kit (DCFH-DA), immunofluorescence staining, q-PCR and flow cytometry. The effects of OST on macrophage polarization and inflammatory responses were investigated by immunoblotting of proteins. Graphpad prism 8.0 software package was used for statistical analysis of the data. RESULTS: CCK-8 results showed that OST was not significantly cytotoxic to RAW264.7 at less than 25 μmol/L, immunofluorescence and q-PCR results showed that OST at 6.25, 12.5 and 25 μmol/L inhibited the expression of inflammatory factors in M1 macrophages, and iNOS, TNF-α, CCR7 were reduced in a concentration-dependent manner, and effectively upregulated the expression of M2 inflammatory factors IL-10, Arg-1 and CD206. Flow cytometry showed that OST effectively inhibited the expression of LPS-induced M1 marker CD86 in macrophages. CONCLUSIONS: OST can regulate lipopolysaccharide-induced M1 macrophages polarization and reduce inflammatory reaction.

Key words: Osthole, Inflammatory bone disease, Macrophage polarization, Lipopolysaccharide

中图分类号:

R318.01

王菲菲, 饶定成, 金思宇, 杨沛鸣, 周永梅, 万光勇. 蛇床子素对脂多糖诱导的巨噬细胞极化和炎症反应的影响[J]. 上海口腔医学, 2022, 31(3): 255-259.

WANG Fei-fei, RAO Ding-cheng, JIN Si-yu, YANG Pei-ming, ZHOU Yong-mei, WAN Guang-yong. The effect of osthole on lipopolysaccharide-induced macrophages polarization and inflammatory reaction[J]. Shanghai Journal of Stomatology, 2022, 31(3): 255-259.

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蛇床子素对脂多糖诱导的巨噬细胞极化和炎症反应的影响

The effect of osthole on lipopolysaccharide-induced macrophages polarization and inflammatory reaction

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