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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IC50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IL-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 effect of Berberine (Ber) on the secretion of inflammatory cytokines TNF-α and TNF-α in RAW264.7 cells induced by lipopolysaccharide (LPS) and interleukin-4 (IL-4) Interleukin (IL-10), to investigate the Ber on the polarization of macrophages. Methods The effect of berberine on the proliferation of RAW264.7 cells was determined by MTT assay. The concentration of berberine below the IC50 was chosen as the intervention dose. The effect of berberine on the proliferation of RAW264.7 cells was detected by enzyme-linked immunosorbent assay (ELISA) And IL-4 induced by different state RAW264.7 cells secrete TNF-α, IL-10. Results The IC50 of berberine to RAW264.7 cells was between 400 ~ 800μmol·L-1. The secretion of pro-inflammatory cytokines TNF-α in RAW264.7 cells was significantly increased by LPS, and Ber (5,10,20 μmol·L-1, respectively) was significantly inhibited (all P <0.01 ), While LPS-stimulated IL-10 secretion was significantly increased (P <0.05), while Ber only showed inhibitory effect at 20μmol·L-1 dose. Under the action of IL-4, the secretion of anti-inflammatory cytokine IL-10 in RAW264.7 cells was significantly increased, Ber (5,10 and 20μmol·L-1 respectively) showed a significant inhibitory effect in a dose-dependent manner (All P <0.01). The secretion of TNF-α was significantly decreased by IL-4 (P <0.05). The effect of Ber on the secretion of TNF-α was dose-dependent. Conclusion Ber can inhibit the secretion of TNF-α, a proinflammatory cytokine induced by LPS, and inhibit the secretion of IL-10 induced by IL-4, suggesting that Ber may play a regulatory role in the polarization of macrophages.