Endothelin-1 increases cholinergic nerve-mediated contraction of human bronchi via tachykinin synthesis induction.
1. In some asthmatics, muscarinic receptor antagonists are effective in limiting bronchoconstrictor response, suggesting an abnormal cholinergic drive in these subjects. There is a growing body of evidences indicating that cholinergic neurotransmission is also enhanced by endothelin-1 (ET-1) in rabbit bronchi, mouse trachea and in human isolated airway preparations. 2. We investigated the role of secondary mediators in ET-1 induced potentiation of cholinergic nerve-mediated contraction in human bronchi, in particular the possible role of neuropeptides in this phenomenon. 3. Bronchial tissues after endothelin treatment were exposed to a standard electrical field stimulation (EFS) (30% of EFS 30 Hz)-induced contraction. In addition, in some experiments, preparations were treated with a tachykinin NK(2) receptor antagonist and subsequently exposed to the same protocol. HPLC and RIA were performed on organ bath fluid samples. Moreover, the human bronchi were used for the beta-PPT (preprotachykinin) mRNA extraction and semiquantitative reverse transcription polymerase chain reaction (RT - PCR), prior to and 30-40 min following ET-1 challenge. 4. The selective tachykinin NK(2) receptor antagonist, SR48968, was effective to reduce ET-1 potentiation of EFS mediated contraction. HPLC or RIA showed significant increased quantities of NKA in organ bath effluents after EFS stimulation in bronchi pretreated with ET-1. Finally, beta-PPT mRNA level after stimulation of bronchi with ET-1 was increased about 2 fold respect to control untreated bronchi. 5. In conclusion, this study demonstrated that, at least in part, the ET-1 potentiation of cholinergic nerve-mediated contraction is mediated by tachykinin release, suggesting that in addition to nerves, several type of cells, such as airway smooth muscle cell, may participate to neuropeptide production.
D'Agostino
B
B
Department of Experimental Medicine, Section of Pharmacology 'Leonardo Donatelli' Faculty of Medicine and Surgery, 2nd University of Naples, Italy. bruno.dagostino@unina2.it
Advenier
C
C
Falciani
M
M
Gallelli
L
L
Marrocco
G
G
Piegari
E
E
Filippelli
A
A
Rossi
F
F
D000109:Acetylcholine / Q000494:pharmacology; D000368:Aged; D001549:Benzamides / Q000494:pharmacology; D001980:Bronchi / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D002799:Cholinergic Fibers / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D019332:Endothelin-1 / Q000494:pharmacology*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D009119:Muscle Contraction / Q000187:drug effects*; D015288:Neurokinin A / Q000378:metabolism; D010880:Piperidines / Q000494:pharmacology; D011498:Protein Precursors / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011863:Radioimmunoassay; D018041:Receptors, Neurokinin-2 / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013373:Substance P / Q000378:metabolism; D015320:Tachykinins / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D014665:Vasodilator Agents / Q000494:pharmacology
D001549:Benzamides; D019332:Endothelin-1; D010880:Piperidines; D011498:Protein Precursors; D012333:RNA, Messenger; D018041:Receptors, Neurokinin-2; D015320:Tachykinins; D014665:Vasodilator Agents; C044206:preprotachykinin; D013373:Substance P; C073839:SR 48968; D015288:Neurokinin A; D000109:Acetylcholine