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目的:探讨红霉素对哮喘大鼠磷酸化P38及Th2类细胞因子的影响。方法:分离培养正常对照组、哮喘组、红霉素处理组大鼠的外周血单核细胞(PBMCs)。分别采用ELISA方法检测细胞上清液中IL-4、IL-5、IL-13浓度。逆转录聚合酶链反应(RT-PCR)、Western blotting技术检测PBMCs P38mRNA、P38磷酸化蛋白的表达。结果:哮喘组PB-MCs P38mRNA、P38磷酸化蛋白表达水平及细胞上清液中IL-4、IL-5、IL-13的浓度较正常对照组显著升高(P<0 .05) ,红霉素处理组P38 mRNA、磷酸化蛋白表达水平及细胞上清液中IL-4、IL-5、IL-13的浓度较哮喘组显著降低(P<0 .05)。通过直线相关分析发现,PBMCs P38磷酸化蛋白与细胞上清液中IL-4、IL-5、IL-13的表达水平之间均呈显著正相关(r分别=0 .75、0 .82、0 .70 ,P均<0 .05)。结论:P38可能参与支气管哮喘的病理生理过程。红霉素可能通过作用于P38这一靶点发挥其免疫调节作用。
Objective: To investigate the effect of erythromycin on phosphorylated P38 and Th2 cytokines in asthmatic rats. Methods: Peripheral blood mononuclear cells (PBMCs) of rats in normal control, asthma and erythromycin groups were isolated and cultured. The concentrations of IL-4, IL-5 and IL-13 in the cell supernatant were detected by ELISA. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of P38mRNA and P38 phosphorylation protein in PBMCs. Results: The expression of PBMCs P38mRNA, P38 phosphorylation protein and IL-4, IL-5 and IL-13 in asthma group were significantly higher than those in control group (P <0.05) The levels of P38 mRNA, phosphorylated protein and IL-4, IL-5 and IL-13 in the supernatant of the untreated group were significantly lower than those in the asthma group (P <0.05). By linear correlation analysis, there was a significant positive correlation between PBMCs P38 phosphorylation protein and IL-4, IL-5, IL-13 expression in cell supernatant (r = 0.75,0.82, 0 .70, all P <0 .05). Conclusion: P38 may be involved in the pathophysiological process of bronchial asthma. Erythromycin may exert its immunomodulatory effect through acting on the target of P38.