Erythromycin Suppresses The Activation Of Nuclear Factor-kappa B Induced By TNF-α In Human Bronchial Epithelial Cells

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 (COPD) . Methods:Human bronchial epithelial cell line 16HBE was cultured with 10% serum DMEM(high glucose).The cells were randomly divided into eight groups such as 1. blank control group; 2. tumor necrosis factor (TNF- a ) (20ng/ml,16h); 3.EM(0.3ug/ml, 24h)+TNF-a (20ng/ml, 16h); 4.EM(3ug/ml, 24h)+TNF-a (20ng/ml, 16h); 5.EM(30ug/ml, 24h)+TNF- a (20ng/ml, 16h); 6.EM(0.3ug/ml, 48h)+TNF-a(20ng/ml, 16h); 7.EM(3ug/ml, 48h)+TNF- a (20ng/ml, 16h); 8.EM(30ug/ml, 48h)+TNF- a (20ng/ml, 16h).The cells were incubated with various concentrations of EM for 24-48 hours before the stimulation by TNF- a . Then total cellular RNA was extracted to detect the expression of interleukin-8 (IL-8) mRNA and intercellular adhesion molecule 1(ICAM-1) mRNA by applying reverse transcription- polymerase chain reaction (RT-PCR) . And cellular nuclear proteins was isolated to detect the activity of nuclear factor-kappa B (NF- k B) by applying electrophoresis mobility shift assay (EMSA). Results:1. The activation of NF- k B in human bronchial epithelial cell line 16HBE induced by TNF- a was inhibited by the preincubation with erythromycin in dose-dependent and time-dependent fashion.2. IL-8mRNA was increased by the addition of TNF- a , which was inhibited by the preincubation with erythromycin.3. ICAM-lmRNA was increased by the addition of TNF- a ,which was inhibited by the preincubation with erythromycin in dose-dependent and time-dependent fashion.Conclusion:1. Our results indicate that IL-8mRNA and ICAM-lmRNA were increased by the stimulation of TNF- a , which may be regulated by the activation of NF-k B induced by TNF- a in human bronchial epithelial cells.2. Our results indicate that erythromycin modify inflammation presumably by suppressing IL-8mRNA and ICAM-lmRNA and that erythromycin might affect the expression of these genes through suppressing the activation of NF-k. B in human bronchial epithelial cells.