Endothelium-dependent contractile actions of proteinase-activated receptor-2-activating peptides in human umbilical vein: release of a contracting factor via a novel receptor

Citation
M. Saifeddine et al., Endothelium-dependent contractile actions of proteinase-activated receptor-2-activating peptides in human umbilical vein: release of a contracting factor via a novel receptor, BR J PHARM, 125(7), 1998, pp. 1445-1454
Citations number
30
Categorie Soggetti
Pharmacology & Toxicology
Journal title
BRITISH JOURNAL OF PHARMACOLOGY
ISSN journal
00071188 → ACNP
Volume
125
Issue
7
Year of publication
1998
Pages
1445 - 1454
Database
ISI
SICI code
0007-1188(199812)125:7<1445:ECAOPR>2.0.ZU;2-3
Abstract
1 The contractile actions of the proteinase-activated receptor-2-activating peptides (PAR(2)APs), SLIGRL-NH2 (SL-NH2), SLIGKV-NH2 (KV-NH2), trans-cinn amoyl-LIGRLO-NH2 (tc-NH2), and the PAR(1)-AP, TFLLR-NH2 (TF-NH2) as well as trypsin and thrombin were studied in endothelium-denuded and intact human umbilical vein (HUV) ring preparations. 2 In HUV rings with, but not without an intact endothelium, PAR(2)APs cause d a concentration-dependent contractile response, whereas LSIGRL-NH2 trypsi n and PAR(1)APs were inactive. The contractile response was not affected by the endothelin ETA receptor antagonist, BQ123, the cyclooxygenase inhibito r, indomethacin, the leukotriene synthesis inhibitor, MK886, or the epoxyge nase/P450 inhibitor, SKF-525A. Other pharmacological antagonists (prazosin, Losartan(R)) were similarly inactive. 3 The order of potencies of the PAR(2)APs to cause a contraction in the end othelium-intact preparation was: SL-NH2> >KV-NH(2)greater than or equal to tc-NH2. 4 Using an endothelium-free rat aorta ring as a reporter tissue, surrounded with endothelium-intact HUV as a donor tissue in a 'sandwich assay,' we al so monitored the ability of SL-NH2, TF-NH2, trypsin and thrombin to release either contractile (EDCF) or relaxant (EDRF) factors. 5 In the 'sandwich assay' done in the presence of L-NAME (0.1 mh I), the en dothelium-intact HUV tissue (but not endothelium-denuded HUV) released a co ntractile factor (EDCF) in response to SL-NH2 (50 mu M) but not to trypsin or LSIGRL-NH2. The SL-NH2-mediated release/action of the EDCF was not affec ted by BQ123, indomethacin, MK886 or SKF-525A. 6 In the 'sandwich assay', trypsin (4-10 nM), SL-NH2, KV-NH2 and tc-NH2 cau sed the release of a relaxant activity (EDRF) from the endothelium-intact ( but not the denuded) HUV preparation. The release of EDRF was blocked by 0. 1 mM (omega)nitro-L-arginine-methylester (L-NAME). Neither thrombin (10 u m l(-1), 100 nM) nor TF-NH2 (50 mu M) were active in this EDRF-release assay. 7 The relative potencies of the PAR(2) agonists for causing the release of EDRF in the HUV sandwich assay were: trypsin > >SL-NH2> >tc-NH2>KV-NH2. Thi s order of potencies differed from the one observed for the same agonists i n the HUV contraction assay (above) and in an intracellular calcium signall ing assay, conducted with cloned human PAR(2) that was expressed in culture d rat kidney KNRK cells: trypsin > > SL-NH2 = tc-NH2 > KV-NH2. 8 We conclude that PAR(2)APs (but not PAR(1)APs) via a receptor distinct fr om PAR(2), can cause a contractile response in endothelium-intact HUV tissu e via the release of a diffusable EDCF, that is different from previously r ecognized smooth muscle agonists (e.g. prostanoid metabolites, endothelin, noradrenaline, angiotensin-II, acetylcholine).