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目的摸索利用IFN-γ诱导THP-1源巨噬细胞活化的方法,筛选用于检测巨噬细胞活化状态的最佳细胞因子和最佳检测时间。方法 PMA(佛波酯)诱导THP-1分化成为巨噬细胞,两次加入IFN-γ诱导巨噬细胞活化(两步法),ELISA法检测不同浓度和不同刺激时间培养上清中细胞因子TNF-α、IL-10、IL-12、IL-23的水平。结果 IFN-γ诱导的巨噬细胞可大量分泌TNF-α,分泌量大于1 000 ng/mL,而IL-10、IL-12、IL-23分泌量少或未见明显分泌;两次IFN-γ诱导巨噬细胞活化的最低浓度均为2.5 ng/mL,初次最佳刺激时间为8~16 h,再次最佳刺激6~18 h。结论 IFN-γ可诱导巨噬细胞活化,两次IFN-γ刺激的效果比单次刺激效果好,TNF-α可作为巨噬细胞活化的理想检测指标。
Objective To explore the method of IFN-γ-induced activation of THP-1-derived macrophages and to screen for the best cytokines and optimal detection time for detecting macrophage activation status. Methods PMA (phorbol ester) induced THP-1 to differentiate into macrophages, twice with IFN-γ to induce macrophage activation (two-step method). The levels of cytokines TNF -α, IL-10, IL-12, IL-23. Results The secretion of TNF-α by IFN-γ-induced macrophages was greater than 1 000 ng / mL, while the secretions of IL-10, IL-12 and IL- The lowest concentration of γ-induced macrophage activation was 2.5 ng / mL, the best initial stimulation time was 8-16 h, and the best stimulation was 6-18 h. Conclusion IFN-γ can induce the activation of macrophages. The effect of two IFN-γ stimulation is better than that of single stimulation. TNF-α can be used as an ideal marker of macrophage activation.