基础医学与临床 ›› 2019, Vol. 39 ›› Issue (5): 646-651.

• 研究论文 • 上一篇    下一篇

小檗碱促进巨噬细胞系RAW264.7由M1促炎表型向M2抗炎表型极化

王青竹1,石婧1,刘琴1,杨黎星1,郭磊1,叶菜英2,张德昌1   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院基础医学研究所药理室
  • 收稿日期:2019-01-10 修回日期:2019-03-15 出版日期:2019-05-05 发布日期:2019-04-24
  • 通讯作者: 郭磊 E-mail:leiguo@ibms.cams.cn
  • 基金资助:
    国家自然科学基金面上项目;中国医学科学院医学与健康科技创新工程

Berberine promotes M1 proinflammatory phenotype to M2 anti-inflammatory phenotype polarization in macrophage cell line RAW264.7

  • Received:2019-01-10 Revised:2019-03-15 Online:2019-05-05 Published:2019-04-24

摘要: 目的 探讨小檗碱(BBR)对 RAW264.7 巨噬细胞极化分型的影响。方法 将 RAW264.7 细胞分为对照组、高脂和炎性反应模型组(以100 μg/L 脂多糖和100 μg/L 聚合型低密度脂蛋白刺激) 和小檗碱低、中、高浓度(5、10、20 μmol/L)处理组。用ELISA和流式细胞计量术检测巨噬细胞两种极化分型( M1 型和 M2 型)相关标志物TNF-α、MCP-1、CD86、IL-4、IL-13、TGF-β1和CD206的表达,以观察细胞极化。用Western blot和RT-qPCR检测载脂蛋白 E(apoE)、极低密度脂蛋白受体(VLDL)或载脂蛋白E受体2(apoER2)的表达。结果 BBR引起M1型巨噬细胞相应标志物TNF-α、MCP-1和CD86表达减少(P<0.05); M2型巨噬细胞相应标志物IL-4、TGF-β1和CD206表达增加。同时也能够促进RAW264.7细胞内apoE的蛋白表达和VLDL的mRNA表达(P<0.001)。结论 BBR可能促进apoE与VLDL的结合,进而诱导RAW264.7从M1促炎表型向M2抗炎表型极化。

关键词: 关键词:小檗碱, 巨噬细胞表型, 炎性反应, 载脂蛋白

Abstract: Objective To test the effect of berberine (BBR) on polarization of RAW264.7 mouse macrophages. Methods RAW264.7 cells were divided into control group, high lipid- and inflammatory-model group (stimulated with 100μg/L lipopolysaccharide and aggregated LDL), BBR 5 μmol/L group, BBR 10 μmol/L group and BBR 20 μmol/L group. ELISA and flow cytometry were used to assess specific cell markers (TNF-α, MCP-1, CD86, IL-4, IL-13, TGF-β1 and CD206) to define M1 and M2 polarization. Protein expression of apolipoprotein E (apoE) and very-low-density lipoprotein receptor (VLDL-R) or mRNA expression of apoE receptor-2 (apoER2) were detected by Western blot and RT-qPCR,respetively. Results BBR decreased M1 macrophage-specific cell markers TNF-α and MCP-1 levels (P<0.05)and downregulated the expressions of CD86. In contrast, M2 macrophage-specific cell markers IL-4, TGF-β1 and CD206were upregulated by BBR. BBR could promote the protein expression of apoE and the mRNA expression of VLDL (P<0.001). Conclusions BBR treatment may shift RAW264.7 mouse macrophage to antiinflammatory M2 phenotype by promoting the binding of apoE to VLDLR.

Key words: Key words Berberine, macrophage phenotype, inflammation, apolipoprotein

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