Milara Javier, Morell Anselm, Ballester Bea, Sanz Celia, Freire Jose, Qian Xiaozhong, Alonso-Garcia Maggie, Morcillo Esteban, Cortijo Julio
Clinical Research Unit, University General Hospital Consortium, Valencia, Spain.
CIBERES, Health Institute Carlos III, Valencia, Spain.
Respir Res. 2015 Feb 5;16(1):12. doi: 10.1186/s12931-015-0179-5.
Chronic obstructive pulmonary disease (COPD) is characterised by chronic pulmonary inflammation punctuated by periods of viral exacerbations. Recent evidence suggests that the combination of roflumilast with corticosteroids may improve the compromised anti-inflammatory properties of corticosteroids in COPD. We analyzed differential and combination anti-inflammatory effects of dexamethasone and roflumilast N-oxide in human bronchial epithelial cells (HBECs) stimulated with viral toll like receptor (TLR) agonists.
Lung tissue and HBECs were isolated from healthy (n = 15), smokers (n = 12) and smokers with COPD (15). TLR3 expression was measured in lung tissue and in HBECs. IL-8 secretion was measured in cell cultures after TLR3 stimulation with poly I:C 10 μg/mL.
We found that TLR3 expression was increased by 1.95 fold (protein) and 2.5 fold (mRNA) in lung tissues from smokers with COPD and inversely correlated with lung function. The TLR3 agonist poly I:C 10 μg/mL increased the IL-8 release in HBECs that was poorly inhibited by dexamethasone in smokers (24.5%) and smokers with COPD (21.6%). In contrast, roflumilast showed similar inhibitory effects on IL-8 release in healthy (58.8%), smokers (56.6%) and smokers with COPD (50.5%). The combination of roflumilast N-oxide and dexamethasone showed additive inhibitory effects. Mechanistically, roflumilast N-oxide when combined with dexamethasone increased the expression of MKP1, and enhanced the inhibitory effects on phospho-p38, AP1 and NFκB activities which may explain the additive anti-inflammatory effects.
Altogether, our data provide in vitro evidence for a possible clinical utility to add roflumilast on top of inhaled corticosteroid in COPD.
慢性阻塞性肺疾病(COPD)的特征是慢性肺部炎症,并伴有病毒感染加重期。最近的证据表明,罗氟司特与皮质类固醇联合使用可能改善COPD中皮质类固醇受损的抗炎特性。我们分析了地塞米松和罗氟司特N-氧化物在病毒Toll样受体(TLR)激动剂刺激的人支气管上皮细胞(HBECs)中的差异抗炎作用和联合抗炎作用。
从健康人(n = 15)、吸烟者(n = 12)和患有COPD的吸烟者(15)中分离肺组织和HBECs。检测肺组织和HBECs中TLR3的表达。在用10μg/mL聚肌胞苷酸刺激TLR3后,检测细胞培养物中IL-8的分泌。
我们发现,患有COPD的吸烟者肺组织中TLR3表达增加了1.95倍(蛋白质)和2.5倍(mRNA),且与肺功能呈负相关。10μg/mL的TLR3激动剂聚肌胞苷酸增加了HBECs中IL-8的释放,地塞米松对吸烟者(24.5%)和患有COPD的吸烟者(21.6%)中IL-8释放的抑制作用较弱。相比之下,罗氟司特对健康人(58.8%)、吸烟者(56.6%)和患有COPD的吸烟者(50.5%)中IL-8释放具有相似的抑制作用。罗氟司特N-氧化物与地塞米松联合使用具有相加的抑制作用。从机制上讲,罗氟司特N-氧化物与地塞米松联合使用时会增加MKP1的表达,并增强对磷酸化p38、AP1和NFκB活性的抑制作用,这可能解释了相加的抗炎作用。
总之,我们的数据提供了体外证据,表明在COPD中,在吸入皮质类固醇基础上加用罗氟司特可能具有临床应用价值。