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