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

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。

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

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.