Thrombin stimulates release of endothelin and vasopressin, but not substance P, from isolated rabbit tracheal epithelial cells

doi:10.1016/0014-2999(93)90575-3

European Journal of Pharmacology

Volume 230, Issue 3, 19 January 1993, Pages 367-370

https://doi.org/10.1016/0014-2999(93)90575-3 Get rights and content

Abstract

Release of endothelin, vasopressin and substance P from isolated rabbit tracheal epithelial cells was monitored after incubation for 2 h with thrombin (10 U/ml) in the presence and absence of cycloheximide (100 μg/ml) and 2-nitro-4-carboxyphenyl-N, N-diphenylcarbamate (NCDC, 0.6 mM). Thrombin stimulated the release of endothelin and vasopressin. Inhibition of the thrombin-stimulated release of endothelin and vasopressin by cycloheximide (44and 56%, respectively) and NCDC (59 and 88%, respectively) indicates that the release of these peptides is at least partially dependent on the ability of thrombin to stimulate protein synthesis and activate the phospholipase C pathway. Substance P, which is also present in tracheal epithelial cells, was not released by thrombin.

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The characteristics of endothelin (ET) release from guinea-pig tracheal epithelium were investigated, including examination of the effects of several pro-inflammatory mediators. In confluent cultured guinea-pig tracheal epithelial cells (GPTECs) there was a time-dependent basal release of immunoreactive ET (ir-ET) from 4–48 h. Basal ir-ET release from GPTECs was unaffected by the peptidase inhibitors, thiorphan (10 μm), benzamidine (1 mm), pepstatin-A (30 μm), aprotinin (1 μg/ml), bacitracin (20 μg/ml) or leupeptin (50 μm), but was inhibited by phosphoramidon, the neutral metalloprotease inhibitor (IC₅₀=16.8 μm), or the calcium chelator, EGTA (10 mm). There was little ir-ET release 1 day after placing GPTECs in culture, although appreciable release (>10-fold higher) was detected on days 5 and 7. No significant release of ir-ET was demonstrated from intact guinea-pig trachea. Human thrombin (0.1–10 U/ml), LPS (0.3–10 ng/ml) and the phorbol ester, phorbol 12-myristate-13-acetate (0.1 nm–1 μm), significantly increased ir-ET release, whereas TNF-α (0.1–10 ng/ml), RANTES (0.1–100 nm), IL-1 (0.01–10 ng/ml), bradykinin (1 nm–10 μm), CGRP (0.01 nm–1 μm), PDGF (0.1–3 ng/ml), Sar⁹,Met(O₂)¹¹-Sub P, Nle¹⁰-NKA 4–10 and senktide (selective NK-1, NK-2 and NK-3 receptor agonists, respectively; 1 nm–10 μm), LTD₄(1 nm–10 μm) or major basic protein (10 nm–1 μm) were without stimulatory effect. The results indicate that the enzyme responsible for the basal release of ET from cultured GPTECs is a Ca²⁺-dependent, phosphoramidon-sensitive, neutral metalloprotease. Furthermore, normally there is minimal ET release from guinea-pig airway epithelium but this can be increased markedly by culturing the cells to confluence, and by select pro-inflammatory mediators.

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Short communicationThrombin stimulates release of endothelin and vasopressin, but not substance P, from isolated rabbit tracheal epithelial cells

Abstract

Biochem. Biophys. Res. Commun.

Eur. J. Pharmacol.

Eur. J. Pharmacol.

Inositol phosphates and cell signalling

Nature

Chronic hypoxia changes the ratio of endothelin to ATP release from rat aortic endothelial cells exposed to high flow

Thrombin and histamine activate phospholipase C in human endothelial cells via a phorbol ester-sensitive pathway

J. Cell. Physiol.

The respiratory epithelium releases a smooth muscle relaxing factor

Chest

Role of epithelium in agonist-induced contractile responses of guinea-pig trachealis: influence of the surface through which drug enters the tissue

Br. J. Pharmacol.

Damage of the airway epithelium and bronchial reactivity in patients with asthma

Am. Rev. Respir. Dis.

Short communication
Thrombin stimulates release of endothelin and vasopressin, but not substance P, from isolated rabbit tracheal epithelial cells