Endothelium-dependent contractile actions of proteinase-activated receptor-2-activating peptides in human umbilical vein

Release of a contracting factor via a novel receptor

Mahmoud Saifeddine, Samir S. Roy, Bahjat Al-Ani, Christopher Triggle, Morley D. Hollenberg

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

1. The contractile actions of the proteinase-activated receptor-2-activating peptides (PAR2APs), SLIGRL-NH2 (SL-NH2), SLIGKV-NH2 (KV-NH2), trans-cinnamoyl-LIGRLO-NH2 (tc-NH2), and the PAR1-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, PAR2APs caused a concentration-dependent contractile response, whereas LSIGRL-NH2 trypsin and PAR1APs were inactive. The contractile response was not affected by the endothelin ETA receptor antagonist, BQ123, the cyclooxygenase inhibitor, indomethacin, the leukotriene synthesis inhibitor, MK886, or the epoxygenase/P450 inhibitor, SKF-525A. Other pharmacological antagonists (prazosin, Losartan®) were similarly inactive. 3. The order of potencies of the PAR2APs to cause a contraction in the endothelium-intact preparation was: SL-NH2 >> KV-NH2 ≥ 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 also 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 mM), the endothelium-intact HUV tissue (but not endothelium-denuded HUV) released a contractile factor (EDCF) in response to SL-NH2 (50 μM) but not to trypsin or LSIGRL-NH2. The SL-NH2-mediated release/action of the EDCF was not affected by BQ123, indomethacin, MK886 or SKF-525A. 6. In the 'sandwich assay', trypsin (4-10 nM), SL-NH2, KV-NH2 and tc-NH2 caused 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 (ω)nitro-L-arginine-methylester (L-NAME). Neither thrombin (10 u ml-1, 100 nM) nor TF-NH2 (50 μM) were active in this EDRF-release assay. 7. The relative potencies of the PAR2 agonists for causing the release of EDRF in the HUV sandwich assay were: trypsin >> SL-NH2 >> tc-NH2 > KV-NH2. This order of potencies differed from the one observed for the same agonists in the HUV contraction assay (above) and in an intracellular calcium signalling assay, conducted with cloned human PAR2 that was expressed in cultured rat kidney KNRK cells: trypsin >> SL-NH2 = tc-NH2 > KV-NH2. 8. We conclude that PAR2APs (but not PAR1APs) via a receptor distinct from PAR2, can cause a contractile response in endothelium-intact HUV tissue via the release of a diffusable EDCF, that is different from previously recognized smooth muscle agonists (e.g. prostanoid metabolites, endothelin, noradrenaline, angiotensin-II, acetylcholine).

Original languageEnglish
Pages (from-to)1445-1454
Number of pages10
JournalBritish Journal of Pharmacology
Volume125
Issue number7
Publication statusPublished - 1998
Externally publishedYes

Fingerprint

PAR-2 Receptor
Umbilical Veins
Endothelium
Trypsin
Peptides
Thrombin
Proadifen
NG-Nitroarginine Methyl Ester
Indomethacin
Calcium Signaling
Losartan
Cyclooxygenase Inhibitors
seryl-leucyl-isoleucyl-glycyl--arginyl-leucinamide
Leukotrienes
Prazosin
Endothelins
Angiotensin II
Acetylcholine
Prostaglandins
Smooth Muscle

Keywords

  • Endothelium-derived contracting factor
  • PAR
  • PAR
  • Proteinase-activated receptor
  • Thrombin
  • Trypsin
  • Umbilical vein

ASJC Scopus subject areas

  • Pharmacology

Cite this

Endothelium-dependent contractile actions of proteinase-activated receptor-2-activating peptides in human umbilical vein : Release of a contracting factor via a novel receptor. / Saifeddine, Mahmoud; Roy, Samir S.; Al-Ani, Bahjat; Triggle, Christopher; Hollenberg, Morley D.

In: British Journal of Pharmacology, Vol. 125, No. 7, 1998, p. 1445-1454.

Research output: Contribution to journalArticle

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T1 - Endothelium-dependent contractile actions of proteinase-activated receptor-2-activating peptides in human umbilical vein

T2 - Release of a contracting factor via a novel receptor

AU - Saifeddine, Mahmoud

AU - Roy, Samir S.

AU - Al-Ani, Bahjat

AU - Triggle, Christopher

AU - Hollenberg, Morley D.

PY - 1998

Y1 - 1998

N2 - 1. The contractile actions of the proteinase-activated receptor-2-activating peptides (PAR2APs), SLIGRL-NH2 (SL-NH2), SLIGKV-NH2 (KV-NH2), trans-cinnamoyl-LIGRLO-NH2 (tc-NH2), and the PAR1-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, PAR2APs caused a concentration-dependent contractile response, whereas LSIGRL-NH2 trypsin and PAR1APs were inactive. The contractile response was not affected by the endothelin ETA receptor antagonist, BQ123, the cyclooxygenase inhibitor, indomethacin, the leukotriene synthesis inhibitor, MK886, or the epoxygenase/P450 inhibitor, SKF-525A. Other pharmacological antagonists (prazosin, Losartan®) were similarly inactive. 3. The order of potencies of the PAR2APs to cause a contraction in the endothelium-intact preparation was: SL-NH2 >> KV-NH2 ≥ 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 also 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 mM), the endothelium-intact HUV tissue (but not endothelium-denuded HUV) released a contractile factor (EDCF) in response to SL-NH2 (50 μM) but not to trypsin or LSIGRL-NH2. The SL-NH2-mediated release/action of the EDCF was not affected by BQ123, indomethacin, MK886 or SKF-525A. 6. In the 'sandwich assay', trypsin (4-10 nM), SL-NH2, KV-NH2 and tc-NH2 caused 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 (ω)nitro-L-arginine-methylester (L-NAME). Neither thrombin (10 u ml-1, 100 nM) nor TF-NH2 (50 μM) were active in this EDRF-release assay. 7. The relative potencies of the PAR2 agonists for causing the release of EDRF in the HUV sandwich assay were: trypsin >> SL-NH2 >> tc-NH2 > KV-NH2. This order of potencies differed from the one observed for the same agonists in the HUV contraction assay (above) and in an intracellular calcium signalling assay, conducted with cloned human PAR2 that was expressed in cultured rat kidney KNRK cells: trypsin >> SL-NH2 = tc-NH2 > KV-NH2. 8. We conclude that PAR2APs (but not PAR1APs) via a receptor distinct from PAR2, can cause a contractile response in endothelium-intact HUV tissue via the release of a diffusable EDCF, that is different from previously recognized smooth muscle agonists (e.g. prostanoid metabolites, endothelin, noradrenaline, angiotensin-II, acetylcholine).

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KW - Endothelium-derived contracting factor

KW - PAR

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KW - Proteinase-activated receptor

KW - Thrombin

KW - Trypsin

KW - Umbilical vein

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