Acetylcholine leads to signal transducer and activator of transcription 1 (STAT-1) mediated oxidative/nitrosative stress in human bronchial epithelial cell line

详细信息    查看全文

• 作者：Mirella Profita ; Giusy Daniela Albano ; Angela Marina Montalbano ; Caterina Di Sano ; Giulia Anzalone ; Rosalia Gagliardo ; Loredana Riccobono ; Anna Bonanno ; Liboria Siena ; Michael Paul Pieper ; Mark Gjomarkaj
• 关键词：Epithelial cell ; Oxidative/nitrosative stress ; STAT-1 ; Acetylcholine
• 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Basis of Disease
• 出版年：December, 2013
• 年：2013
• 卷：1832
• 期：12
• 页码：1949-1958
• 全文大小：1148 K

文摘

The induction of nitric oxide synthase (iNOS) expression via the signal transducer and activator of transcription 1 (STAT-1) is involved in the mechanism of oxidative/nitrosative stress. We investigated whether acetylcholine (ACh) generates oxidative/nitrosative stress in bronchial epithelial cells during airway inflammation of COPD and evaluated the effects of Tiotropium, a once-daily antimuscarinic drug, and Olodaterol, a long-acting 尾2-agonist on these mechanisms. Human bronchial epithelial cells (16-HBE) were stimulated (4 h, 37 掳C) with induced sputum supernatants (ISSs) from healthy controls (HC) (n = 10), healthy smokers (HS) (n = 10) or COPD patients (n = 10), as well as with ACh (from 1 渭M to 100 渭M). The activation of STAT-1 pathway (STAT-1Ser727 and STAT-1Tyr701) and iNOS was evaluated in the cell lysates by Western blot analysis as well as nitrotyrosine levels by ELISA, while reactive oxygen species (ROS) were evaluated by flow cytometry. Finally, the effect of Tiotropium (Spiriva庐) (100 nM), alone or in combination with Olodaterol (1 nM), was tested in this model. ISSs from COPD patients significantly increased the phosphorylation of STAT-1Ser727 and STAT-1Tyr701, iNOS and ROS/Nitrotyrosine when compared with ISSs from HC or HS subjects in 16-HBE cells. Furthermore, synthetic ACh increased all these parameters in stimulated 16HBE when compared with untreated cells. Tiotropium and Olodaterol reduced the oxidative/nitrosative stress generated by ACh and ISSs. We concluded that ACh mediated the oxidative/nitrosative stress involving the STAT-1 pathway activation in human bronchial epithelial cells during COPD. 尾2-Long acting and antimuscarinic drugs, normally used in the treatment of COPD as bronchodilator, might be able to control these cellular events.