克拉霉素抑制气道黏液高分泌细胞内机制的研究

摘要

Objective To investigate the inhibit effect of clarithromycin (CLA) on airway mucous hypersecretion and it's intracellular molecule mechanism.Methods Stimulated human bronchial epithelial cell line HBE16 with neutrophil elastase (NE) to create airway mucous hypersecretion model, interfered the model with CLA and external-signal regulated kinase 1/2 (ERK1/2) specific inhibitor U0126 respectively.Gene transcription and protein expression levels of mucin(MUC)5AC in each group were analyzed by quantitative RT-PCR and ELISA.Phosphorylated epidermal growth factor receptor (p-EGFR), Phosphorylated ERK (p-ERK), Phosphorylated c-jun N-terminal kinase (p-JNK) and Phosphorylated P38 (p-P38) were all analyzed by Western blotting.Results CLA and U0126 reduced the the levels of MUC5AC gene transcription and protein expression in airway mucous hypersecretion model, also the protein expressions of p-ERK.But CLA and U0126 didn't reduce the expressions of p-EGFR, p-JNK and p-P38 obviously.Conclusion CLA could inhibit the intracellular signaling pathway molecule ERK1/2's phosphorylation level thereby reduce the production of MUC5AC in airway epithelial cells, thus might inhibits airway mucous hypersecretion.%目的 研究克拉霉素(CLA)抑制气道黏液高分泌的细胞内分子机制.方法 运用中性粒细胞弹性蛋白酶(NE)刺激人类支气管上皮细胞株HBE16构建气道黏液高分泌模型,予以CLA和信号末端调节激酶1/2(ERK1/2)特异性抑制剂U0126进行干预,采用RT-PCR检测细胞内黏蛋白(MUC)5AC mRNA表达水平,ELISA法检测细胞上清液中MUC5AC蛋白含量,Westernblotting检测细胞内磷酸化表皮生长因子受体(p-EGFR)、磷酸化ERK(p-ERK)、磷酸化c-Jun N端激酶(p.ⅢK)、磷酸化P38(p-P38)的含量.结果 克拉霉素和U0126均能降低气道黏液高分泌模型中MUC5AC基因转录和蛋白表达水平,并且使p-ERK表达水平下降,但对p-EGFR、p-JNK、p-P38的表达无明显作用.结论 克拉霉素能抑制细胞内信号通路分子ERK1/2磷酸化水平,从而减少了气道上皮细胞产生MUC5AC,由此发挥抑制气道黏液高分泌的作用.