Phosphodiesterase 4 inhibition decreases MUC5AC expression induced by epidermal growth factor in human airway epithelial cells
Phosphodiesterase 4 inhibition decreases MUC5AC expression induced by epidermal growth factor in human airway epithelial cells
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DOI：
10.1136/thx.2004.025692
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发表时间：
2005-02-01
期刊：
影响因子：
10
通讯作者：
Morcillo, EJ
中科院分区：
文献类型：
--
作者：
Mata, M;Sarriá, B;Morcillo, EJ
背景：慢性炎症性气道疾病如哮喘和慢性阻塞性肺疾病（COPD）的一个共同病理特征是粘液分泌过多。MUC5AC是在健康气道中表达的主要粘蛋白基因，在哮喘和COPD患者中表达增加。最近的临床试验表明，磷酸二酯酶4 （PDE4）抑制剂可能对慢性阻塞性肺病和哮喘有治疗价值。然而，它们对粘蛋白表达的直接影响很少被研究。方法：在体外培养的人气道上皮细胞（A549）和经表皮生长因子（EGF, 25 ng/ml）刺激的人离体支气管组织中检测MUC5AC mRNA和蛋白的表达。实时RT-PCR检测MUC5AC mRNA， ELISA（细胞裂解液和组织匀浆）、Western blotting（组织匀浆）和免疫组织化学检测MUC5AC蛋白。结果：EGF增加了A549细胞MUC5AC mRNA和蛋白的表达。PDE4抑制剂对EGF诱导的MUC5AC mRNA和蛋白表达产生浓度依赖性抑制，效价（-log IC50）：罗氟司特（类似于7.5）。罗利普兰（类似于6.5）和西洛司特（类似于5.5）。罗氟司特还抑制EGF诱导的A549细胞中磷酸化酪氨酸蛋白、EGF受体以及磷酸化p38-和p44/42-MAPK的表达。在人离体支气管中，EGF诱导的MUC5AC mRNA和蛋白表达被罗氟米司特（1mum）抑制，MUC5AC免疫组化染色呈阳性。结论：选择性抑制PDE4可有效降低EGF诱导的人气道上皮细胞MUC5AC的表达。这一作用可能有助于这类新药治疗黏液高分泌性疾病的临床疗效。
Background: A common pathological feature of chronic inflammatory airway diseases such as asthma and chronic obstructive pulmonary disease (COPD) is mucus hypersecretion. MUC5AC is the predominant mucin gene expressed in healthy airways and is increased in asthmatic and COPD patients. Recent clinical trials indicate that phosphodiesterase type 4 (PDE4) inhibitors may have therapeutic value for COPD and asthma. However, their direct effects on mucin expression have been scarcely investigated.Methods: MUC5AC mRNA and protein expression were examined in cultured human airway epithelial cells (A549) and in human isolated bronchial tissue stimulated with epidermal growth factor (EGF; 25 ng/ml). MUC5AC mRNA was measured by real time RT-PCR and MUC5AC protein by ELISA (cell lysates and tissue homogenates), Western blotting (tissue homogenates) and immunohistochemistry.Results: EGF increased MUC5AC mRNA and protein expression in A549 cells. PDE4 inhibitors produced a concentration dependent inhibition of the EGF induced MUC5AC mRNA and protein expression with potency values (-log IC50): roflumilast (similar to7.5). rolipram (similar to6.5) > cilomilast (similar to5.5). Roflumilast also inhibited the EGF induced expression of phosphotyrosine proteins, EGF receptor, and phospho-p38- and p44/42-MAPK measured by Western blot analysis in A549 cells. In human isolated bronchus, EGF induced MUC5AC mRNA and protein expression was inhibited by roflumilast (1 muM) as well as the MUC5AC positive staining shown by immunohistochemistry.Conclusion: Selective PDE4 inhibition is effective in decreasing EGF induced MUC5AC expression in human airway epithelial cells. This effect may contribute to the clinical efficacy of this new drug category in mucus hypersecretory diseases.