核因子-κB在红霉素抗炎机制的表达

目的：从分子水平上探讨红霉素（EM）的抗炎作用机制,为临床防治慢性阻塞性肺疾病提供新的思路。方法：体外培养人支气管上皮细胞（16HBE）,将细胞随机分为8组,先加入EM干预,后加入肿瘤坏死因子（TNF-α）刺激。分组：①空白对照组;②TNF-α（20μg／L,16h）;③EM（0．3mg／L,24h）＋TNF-α（20μg／L,16h）;④EM（3mg／L,24h）＋TNF-α（20μg／L,16h）;⑤EM（30mg／L,24h）＋TNF-α（20μg／L,16h）;⑥EM（0．3mg／L,48h）＋TNF-α（20μg／L,16h）。然后收集各组细胞分别提取核蛋白,应用电泳迁移率改变分析法（EMSA）检测核因子-κB（NF-κB）的活性。结果：EMSA结果显示,TNF-α刺激人支气管上皮细胞（16HBE）后,细胞内NF-κB表达增高。先加入不同浓度及作用时间的EM,后加入前炎因子TNF-α刺激16HBE,EMSA结果显示各组细胞NF—κB表达均降低,且提示有随着EM浓度增加,作用时间延长,NF-κB表达受抑制愈明显的趋势,但随着EM浓度增加及作用时间延长到一定的范围,NF-κB表达受抑制差别不明显。结论：TNF-α能激活16HBENF-κB,促进炎症的发生发展,在炎症进程中发挥着重要作用。EM能抑制人16HBENF-κB的激活水平,这可能是EM的抗炎作用机制之一。

THE EXPRESSION OF NUCLEAR FACTOR-KAPPA B IN THE ANTI-INFLAMMATORY MOLECULAR MECHANISM OF ERYTHROMYCIN

Objective： To investigate the anti-inflammatory molecular mechanism of erythromycin （EM） in order to find a new way to prevent and treat chronic obstructive pulmonary disease. Methods： Human bronchial epithelial cell line 16HBE was cultured with 10% serum DMEM（high glucose）. The cells were randomly divided into eight groups ： （1）control group;（2）tumor necrosis factor （TNF-α）（20 μg/L, 16 h）;（2）EM （0. 3 mg/L,24 h）＋TNF-α（20 μg/L,16 h）;（4）EM（3 mg/L,24 h）4-TNF-α（20 μg/L,16 h）;（5）EM（30 mg/ L,24 h）＋TNF-α（20 μg/L,16 h） ;（6）EM（0.3 Mg/L,48 h）＋TNF-α（20 μg/L,16 h）. The cells were incubated with various concentrations of EM for 24~48 hours before the stimulation by TNF-α. The cellular nuclear protein was isolated to detect the activity of nuclear factor-kappa B（NF-κB）by applying electrophoresis mobility shift assay （EMSA）. Results： The activation of NF-κB in human bronchial epithelial cell line 16HBE induced by TNF-α was inhibited by the preincubation with erythromycin in dose-dependent and time-dependent fashion. Conclusion： Our results indicate that erythromycin can modify inflammation presumably by suppressing the activation of NF-κB in human bronchial epithelial cells.