Identification of two distinct vasodilator pathways activated by ATP in the mesenteric bed of the rat.

Adenosine 5'-triphosphate (ATP) has important roles in the cardiovascular system, modulating vascular tone by acting as both a vasoconstrictor and a vasodilator. The dilator function of ATP is traditionally thought to be monophasic and mediated primarily by nitric oxide (NO). Here we have identified the endothelium-dependent biphasic nature of ATP-induced vasodilatation of the rat isolated mesenteric bed and investigated the two distinct pathways involved. ATP, at doses of 1x10(-11) to 1x10(-8) moles, induced transient relaxations that were inhibited by the NO synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME: 1x10(-4) M), the soluble guanylyl cyclase inhibitor, 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ: 3x10(-6) M) and KCl (6x10(-2) - 1.2x10(-1) M). At doses upwards of 1x10(-8) moles (1x10(-8) - 3x10(-7) moles), ATP also induced prolonged vasodilatations which were unaltered by L-NAME, L-NAME (1x10(-3) M) and indomethacin (1x10(-5) M), or by ODQ, but were abolished in the presence of KCl. In addition, the cannabinoid CB(1) receptor antagonist SR141716A (1x10(-5) M) was found to inhibit the second prolonged phase of vasodilatation. However, at the concentration used SR141716A is reported to be non-selective. A second CB(1) receptor antagonist, AM251 (1x10(-6) M), had a small but significant inhibitory effect on the second phase of ATP-induced vasodilatation. SR141716A, AM251 and KCl (6x10(-2) - 1.2x10(-1) M) all inhibited anandamide-induced relaxation of the isolated mesenteric bed. These observations demonstrate that ATP stimulates vasodilatation of the mesenteric bed by two distinct mechanisms involving the release of NO and an EDHF. In the absence of better pharmacological tools we can only speculate as to the involvement of an endogenous CB(1) receptor ligand in these responses.

Release of GM-CSF and G-CSF by human arterial and venous smooth muscle cells: differential regulation by COX-2.

In addition to their traditional contractile function, vascular smooth muscle cells can be stimulated under inflammatory conditions to release a range of potent biological mediators. Indeed, we and others have shown that human vascular smooth muscle release the colony stimulating factors (CSF) granulocyte macrophage-CSF (GM-CSF) and granulocyte-CSF (G-CSF) as well as large amounts of prostaglandins following the induction of cyclo-oxygenase-2 (COX-2), when stimulated with cytokines. Here we demonstrate, for the first time, that co-induced COX-2 activity simultaneously suppresses GM-CSF release and potentiates G-CSF release by human vascular cells. Moreover, the differential regulation of GM-CSF and G-CSF release by COX-2 was mimicked by the prostacyclin (PGI(2)) mimetic, cicaprost. These observations suggest that PGI(2), released following the induction of COX-2, differentially regulates the release of GM-CSF (suppresses) and G-CSF (potentiates) from human vascular cells.

Cyclooxygenase-2 regulates granulocyte-macrophage colony-stimulating factor, but not interleukin-8, production by human vascular cells: role of cAMP.

Vascular smooth muscle is now recognized as an important site of mediator generation under inflammatory conditions. Indeed, the release of leukocyte activators, such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-8, by human arterial smooth muscle cells has recently been demonstrated. However, the potential for venous cells to release GM-CSF has not been addressed. We have shown that human vascular smooth muscle cells express the "inflammatory" form of cyclooxygenase (COX), cyclooxygenase-2 (COX-2), when stimulated with cytokines. In some nonvascular cell types, the COX activity has been shown to regulate the release of GM-CSF and IL-8, although the nature of the isoform responsible was not addressed. We show that human venous smooth muscle cells, like their arterial counterparts, release GM-CSF after stimulation with IL-1beta. Similarly, both cell types released IL-8. Under the same conditions, we found that COX-2 activity suppressed GM-CSF, but not IL-8, release by both types of human vascular cells. Moreover, the prostacyclin mimetic, cicaprost, and the cAMP analogue, dibutyryl cAMP, inhibited GM-CSF release from these cells. These observations suggest that COX-2 activity suppresses GM-CSF release via a cAMP-dependent pathway in human vascular cells and illustrates a novel mechanism by which this enzyme can modulate immune and inflammatory events.