小檗碱对LPS、IL-4诱导的小鼠RAW264.7细胞分泌TNF-α、IL-10的影响

Effects of Berberine on TNF-α and IL-10 Secretion Induced by LPS and IL-4 in RAW264.7 Cells

目的观察小檗碱（Berberine，Ber）对脂多糖（LPS）以及白介素-4（IL-4）诱导的RAW264．7细胞株分泌炎症因子中肿瘤坏死因子-α（TNF-α）、抑炎因子白介素（IL-10）的影响，初步探讨Ber对巨噬细胞极化的影响，方法噻唑蓝（MTT）法测定小檗碱对RAW264．7细胞增殖的影响，选取半数抑制浓度（IC50）以下的小檗碱浓度作为干预剂量；酶联免疫法（ELISA）检测小檗碱对LPS以及IL-4不同诱导状态下RAW264．7细胞分泌TNF-α、IL-10的影响。结果小檗碱对RAW264．7细胞的IC铀在400～800μmol·L-1之间。LPS作用下RAW264．7细胞促炎因子TNF-α分泌量明显升高，Ber（剂量分别为5，10，20μmol·L-1）有显著的抑制作用，并有剂量依赖关系（均P〈0．01）；而LPS刺激下细胞IL-10分泌量也明显升高（P〈0．05），而Ber仅在20μmol·L-1剂量下表现抑制作用。在IL-4作用下，RAW264．7细胞抗炎因子IL-10分泌量明显升高，Ber（剂量分别为5，10，20μmol·L-1）有显著的抑制作用，并呈现剂量依赖关系（均P〈0．01）；IL-4作用下TNF-α分泌量明显降低（P〈0．05），Ber对TNF-α分泌量的影响无剂量依赖性。结论Ber对LPS刺激下促炎因子TNF-α的分泌具有抑制作用，对IL-4诱导下1L-10分泌量的升高也有抑制作用，提示Ber在巨噬细胞极化过程中有调节作用．

Objective To observe the effects of berberine on inflammatory eytokine tumor necrosis factor alpha （TNF-a） and anti-inflammatory cytokine interleukin-10 （IL-10） secreted by macrophage RAW264.7 cells afterstimulated by lipopolysaccharide（LPS） and IL-4 respectively, and to discuss preliminarily the effect of berberine on macrophage polarization. Methods The effect of berberine at the concentration under ICs0 on RAW264.7 cells proliferation was detected by MTT assay. Macrophage RAW 264.7 cells were stimulated by LPS and IL-4 respectively, and then the secretion of TNF-α and IL-10 was measured by ELISA. Results IC50 of berberine to RAW264.7 cells was in the range of 400-800 μmol·L-1. The concentration cells after LPS stimulation, and then was inhibited by of TNF-α increased significantly（P〈 0.01） in RAW264.7 berberine at the concentration of 5, 10, 20 μmol·L-1 significantly（P 〈 0.01 ） in a dose-dependent manner. Meanwhile, the concentration of IL-10 was also increased significantly （P 〈 0.05）, and then was inhibited by berberine at 20 μmol·L-1. When RAW264.7 cells were stimulated by IL-4, the concentration of IL-10 was increased and TNF-α was decreased significantly, and berberine at the concentration of 5, 10, 20μmol·L-1had dose-dependent effect only on decreasing IL-10 content. Conclusion Berberine can inhibit RAW264.7 ceils secreting pro-inflammatory cytokine TNF-α when stimulated by LPS, and can inhibit RAW264.7 cells secreting anti-inflammatory cytokine IL-10 when stimulated by IL-4, which suggests that berberine plays a regulation role in macrophage polarization process.