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目的探讨红霉素(EM)对香烟烟雾提取物(CSE)刺激的人巨噬细胞释放白细胞介素-8(IL-8)的影响及其机制。方法体外培养人类单核细胞系U937细胞,用佛波酯将其诱导分化为人巨噬细胞,ELISA试剂盒检测细胞上清液IL-8的质量浓度;凝胶迁移率阻滞实验(EMSA)检测核因子-κB(NF-κB)的活性;蛋白印迹实验(Western blot)检测组蛋白去乙酰化酶1(HDAC1)和NF-κB蛋白表达。结果红霉素预孵育可抑制香烟烟雾诱导的人巨噬细胞上清中IL-8的含量可下调香烟烟雾诱导的人巨噬细胞转录因子NF-κB的活性,可增强香烟烟雾抑制的人巨噬细胞HDAC1蛋白表达。结论红霉素可能通过上调香烟烟雾抑制的HDAC1蛋白表达水平而抑制NF-кB活性,进而抑制炎症介质IL-8释放。
Objective To investigate the effect and mechanism of erythromycin (EM) on the release of interleukin-8 (IL-8) from human macrophages stimulated by cigarette smoke extract (CSE). Methods Human monocytic U937 cells were cultured in vitro and induced to differentiate into human macrophages with phorbol myristate. The concentration of IL-8 in supernatant of the cells was measured by ELISA kit. The results of gel permeation retardation assay (EMSA) Nuclear factor-κB (NF-κB) activity was detected by immunohistochemistry. Histone deacetylase 1 (HDAC1) and NF-κB protein expression were detected by Western blotting. Results Erythromycin preincubation inhibited cigarette smoke-induced IL-8 levels in human macrophage supernatant and reduced cigarette smoke-induced human macrophage transcription factor NF-κB activity, and enhanced human cigarette smoke suppression Macrophage HDAC1 protein expression. Conclusion Erythromycin may inhibit NF-κB activity by up-regulating the HDAC1 protein expression inhibited by cigarette smoke, and then inhibit IL-8 release from inflammatory mediators.