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目的:体外研究雷公藤多甙与白介素-10(IL-10)对人外周血树突细胞表面HLA-DR、CD80抗原表达及IL-12 p40蛋白表达和mRNA转录的影响。方法:采用GM-CSF、IL-4和TNFα体外培养体系获得人外周血树突细胞(DC),HLA-DR和CD80抗原的表达经免疫荧光染色后采用流式细胞仪进行分析,用酶联免疫吸附测定和逆转录聚合酶链反应分别检测IL-12p40蛋白水平和mRNA的转录。结果:雷公藤多甙5-20mg/L显著下调HLA-DR和CD80抗原的表达,IL-10 50-200μg/L能抑制HLA-DR和CD80抗原的表达,并都具有剂量依赖关系;雷公藤多甙5-20mg/L和IL-10 50-200μg/L均能抑制DC分泌IL-12p40蛋白,同时经雷公藤多甙20mg/L和IL-10 200μg/L处理的DC中IL-12 p40 mRNA的表达受到显著抑制。结论:雷公藤多甙和IL-10能通过抑制DC表面分子的表达和IL-12的合成而发挥免疫抑制作用。
OBJECTIVE: To study the effect of Tripterygium wilfordii polyglucoside and interleukin-10 (IL-10) on the expression of HLA-DR and CD80 on human peripheral blood dendritic cells and on the expression of p40 protein and mRNA in IL-12. Methods: Human peripheral blood dendritic cells (DCs) were obtained by using GM-CSF, IL-4 and TNFα in vitro. The expression of HLA-DR and CD80 antigen was analyzed by flow cytometry after immunofluorescence staining. The levels of IL-12p40 protein and mRNA were detected by immunoadsorption assay and reverse transcription-polymerase chain reaction respectively. Results: Tripterygium glycosides 5-20mg / L significantly downregulated the expression of HLA-DR and CD80 antigens, and IL-10 50-200μg / L inhibited the expression of HLA-DR and CD80 antigens in a dose-dependent manner. Tripterygium wilfordii Polyglucoside 5-20mg / L and IL-10 50-200μg / L can inhibit the DC secretion of IL-12p40 protein, and IL-12 p40 in DC treated with tripterygium glycosides 20mg / L and IL-10 200μg / L mRNA expression was significantly inhibited. Conclusion: Tripterygium glycosides and IL-10 can exert immunosuppressive effects by inhibiting the expression of DCs and the synthesis of IL-12.