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

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

目的探讨小檗碱(BBR)对巨噬细胞系RAW264.7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100μg/L脂多糖和100μg/L聚合型低密度脂蛋白刺激)和小檗碱低、中、高浓度(5、10、20μmol/L)处理组。用ELISA和流式细胞计量术检测RAW264.7两种极化分型(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(VLDLR)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VLDLR的mRNA表达(P<0.001)。结论 BBR可能促进apoE与VLDLR的结合,进而诱导RAW264.7从M1促炎表型向M2抗炎表型极化。

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 100 μg/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 expression of CD86. In contrast, M2 macrophage-specific cell markers IL-4, TGF-β1 and CD206 were upregulated by BBR. BBR promoted the protein expression of apoE and the mRNA expression of VLDLR ( P <0.001). Conclusions BBR treatment may shift RAW264.7 mouse macrophage to antiinflammatory M2 phenotype by promoting the binding of apoE to VLDLR.