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目的观察褐藻多糖硫酸酯(Fucoidan)对巨噬细胞RAW264.7体外吞噬活性、细胞因子TNF-α和IL-6分泌,以及Toll样受体4(TLR4)mRNA表达的影响。方法实验分对照组,Fucoidan高、中、低剂量组(浓度分别是200、400和800μg/mL)。药物处理6~48h后,MTT法检测RAW264.7细胞活力;中性红比色法检测细胞吞噬活性;ELISA法检测培养上清中TNF-α和IL-6的分泌水平;实时定量PCR检测Toll样受体4(TLR4)mRNA表达量。结果与对照组相比,Fucoidan显著增强RAW264.7细胞代谢活力和吞噬能力(P<0.01),增加TNF-α和IL-6的分泌,上调TLR4的表达,呈剂量依赖关系。结论 Fucoidan可上调TLR4表达,增强巨噬细胞代谢和吞噬活性,增加TNF-α和IL-6的分泌,具有潜在的调节免疫作用。
Objective To observe the effects of fucoidan on macrophage RAW264.7 in vitro phagocytic activity, cytokines TNF-α and IL-6 secretion and Toll-like receptor 4 (TLR4) mRNA expression. Methods Experimental control group, Fucoidan high, medium and low dose groups (concentrations were 200,400 and 800μg / mL). After 6 to 48 hours of drug treatment, the viability of RAW264.7 cells was detected by MTT assay; the phagocytic activity of the cells was detected by neutral red colorimetric assay; the levels of TNF-α and IL-6 secreted by the supernatant were measured by ELISA; Like receptor 4 (TLR4) mRNA expression. Results Compared with the control group, Fucoidan significantly increased the viability and phagocytosis of RAW264.7 cells (P <0.01), increased the secretion of TNF-α and IL-6 and up-regulated the expression of TLR4 in a dose-dependent manner. Conclusion Fucoidan can up-regulate the expression of TLR4, enhance the metabolism and phagocytosis of macrophages, and increase the secretion of TNF-α and IL-6, which may potentially regulate the immune function.