prostaglandin-h2 and ozagrel

It has been shown that increases in intraluminal flow elicit dilation in venules, but the mediation of response is not yet clarified. We hypothesized that - in addition to nitric oxide (NO) and dilator prostaglandins (PGI(2)/ PGE(2)) - thromboxane A(2) (TxA(2)) contributes to the mediation of flow-induced responses of venules.. Isolated rat gracilis muscle venules (259 +/- 11 microm at 10 mm Hg) dilated as a function of intraluminal flow, which was augmented in the presence of the TxA(2) receptor antagonist SQ 29,548 or the TxA(2) synthase inhibitor ozagrel. In the presence of SQ 29,548, indomethacin or Nomega-nitro-L-arginine methyl-ester decreased flow-induced dilations, whereas in their simultaneous presence dilations were abolished. The selective cyclooxygenase (COX) 1 inhibitor SC 560 reduced, whereas the selective COX-2 inhibitor NS 398 enhanced flow-induced dilations. Immunohistochemistry showed that both COX-1 and COX-2 are present in the wall of venules.. In skeletal muscle venules, increases in intraluminal flow elicit production of constrictor TxA(2), in addition to the dilator NO and PGI(2)/PGE(2), with an overall effect of limited dilation. These mediators are likely to have important roles in the multiple feedback regulation of wall shear stress in venules during changes in blood flow velocity and/or viscosity.

Topics: Animals; Blood Flow Velocity; Blood Viscosity; Bridged Bicyclo Compounds, Heterocyclic; Cyclooxygenase 1; Cyclooxygenase 2; Dinoprostone; Enzyme Inhibitors; Epoprostenol; Fatty Acids, Unsaturated; Hydrazines; Male; Methacrylates; Muscle, Skeletal; NG-Nitroarginine Methyl Ester; Nitric Oxide; Prostaglandin H2; Rats; Rats, Wistar; Regional Blood Flow; Stress, Mechanical; Thromboxane A2; Venules

Stretch-induced contraction of rabbit pulmonary artery depends on endothelium-derived vasoactive prostanoids. We investigated which prostanoid(s) was responsible for the stretch-induced contraction of the artery, and whether integrin was involved in this mechanotransduction process. Stretch increased productions of untransformed prostaglandin H(2), prostaglandin E(2), prostaglandin F(2alpha), and thromboxane A(2) in the pulmonary artery with intact endothelium. A blocking peptide for integrins (RGD peptide) significantly inhibited productions of thromboxane A(2) and prostaglandin F(2alpha), but the peptide did not affect productions of untransformed prostaglandin H(2) and prostaglandin E(2), as well as contraction in response to stretch. SQ29,548, a prostaglandin H(2)/thromboxane A(2) receptor antagonist, inhibited the contractile response to not only stretch but also exogenous prostaglandin H(2). Acetylcholine (up to 30 microM) also contracted the artery in an endothelium-dependent manner. Ozagrel (10 nM-1 microM), an inhibitor of thromboxane synthase, abolished the production of thromboxane A(2), in response to both stretch and acetylcholine, whereas the inhibitor mostly inhibited acetylcholine-induced contraction, but it did not suppress stretch-induced contraction. The results suggested that prostaglandin H(2) and thromboxane A(2), either released from endothelium by mechanical stretch or by acetylcholine, produced contraction of rabbit pulmonary artery in a RGD-independent manner.

Topics: Acetylcholine; Animals; Bridged Bicyclo Compounds, Heterocyclic; Dinoprost; Dinoprostone; Endothelium, Vascular; Fatty Acids, Unsaturated; Hydrazines; In Vitro Techniques; Isometric Contraction; Methacrylates; Oligopeptides; Prostaglandin Antagonists; Prostaglandin H2; Pulmonary Artery; Rabbits; Receptors, Thromboxane A2, Prostaglandin H2; Stress, Mechanical; Thromboxane A2; Thromboxane-A Synthase; Vasoconstriction; Vasodilator Agents

Arachidonic acid (AA) is a potent inducer of platelet aggregation in vitro; this activity is due to its conversion to biologically active metabolites, prostaglandin (PG) endoperoxides and thromboxane A2 (TxA2). PG endoperoxides and TxA, are thought to act on the same receptor; however, at least two isoforms of this receptor have been identified. The aim of our work was to clarify whether endoperoxides and TxA2 activate the same or different receptor subtypes to induce aggregation and calcium movements in human platelets. AA-induced aggregation and calcium rises were still detectable in platelets preincubated with thromboxane synthase inhibitors, which suppress TxA2 formation and induce PGH2 accumulation, suggesting that PG endoperoxides can activate platelets. Exogenously added PGH2 was able to induce aggregation and calcium rises. Pretreatment of platelets with GR32191B or platelet activating factor, which desensitize one of the two receptor subtypes identified in platelets, did not prevent calcium rises induced by endogenously generated or by exogenouly added PGH2, indicating that TxA2 and PG endoperoxides share the same receptor subtype(s) to activate platelets. HEK-293 cells overexpressing either of the two thromboxane receptor isoforms cloned to date (TPalpha and TPbeta) and identified in human platelets, stimulated with PGH2, or with the stable endoperoxide analog U46619, formed inositol phosphates. These data show that endoperoxides and TXA2 mediate their effects on platelets acting on both, and the same, receptor isoform(s).

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Aspirin; Biphenyl Compounds; Blood Platelets; Bridged Bicyclo Compounds, Heterocyclic; Calcium Signaling; Cells, Cultured; Enzyme Inhibitors; Fatty Acids, Unsaturated; Heptanoic Acids; Humans; Hydrazines; Imidazoles; Inositol Phosphates; Kidney; Methacrylates; Phenylacetates; Platelet Activating Factor; Platelet Activation; Prostaglandin H2; Prostaglandins H; Protein Isoforms; Receptors, Thromboxane; Recombinant Fusion Proteins; Sulfonamides; Thromboxane A2; Thromboxane B2; Thromboxane-A Synthase

We investigated the role of endothelium and vasoconstrictor prostanoids in the norepinephrine (NE)-induced vasoconstriction of isolated rat common carotid arteries (CCAs). Isolated CCAs were cannulated with stainless steel cannulae by the cannula inserting method. NE was administered intra- or extraluminally by a single microinjection. For denudation of endothelium, the intimal surface was gently rubbed with a cotton pellet. NE produced dose-related vasoconstricting responses in isolated arteries with intact endothelium. These responses were attenuated by pretreatment with denudation, OKY046, a thromboxane A2 synthesis inhibitor or indomethacin. The residual responses after denudation were further blocked by OKY046 or indomethacin. All NE-induced responses were blocked by a single injection of prazosin. There were no significant differences in responses between intra- and extraluminal administration of NE. These results show that most vasoconstriction induced by NE via alpha1-adrenoceptors in CCAs is dependent on vasoconstrictor prostanoids distributed both in endothelium and vascular smooth muscle.

Topics: Animals; Carotid Artery, Common; Endothelium, Vascular; In Vitro Techniques; Indomethacin; Male; Methacrylates; Norepinephrine; Perfusion; Prostaglandin H2; Prostaglandins; Prostaglandins H; Rats; Rats, Wistar; Receptors, Adrenergic, alpha-1; Thromboxane A2; Vasoconstriction

To clarify the mode of action of a selective thromboxane A2 (TXA2) blockade in platelet reactivity, we examined the effect of (E)-3-[4-(1-imidazolylmethyl) phenyl]-2-propenoic acid hydrochloride (OKY-046), a potent TXA2 synthetase inhibitor, on human platelet aggregation induced by arachidonic acid (1 mM) in the absence and presence of aspirin-treated aortic microsomes containing prostacyclin (PGI2) synthetase activity ex vivo. The production of TXA2 and PGI2 in platelet rich plasma was determined by the amounts of their stable catabolites, TXB2 and 6-keto-PGF1 alpha respectively, measured by radioimmunoassay. In the absence of aortic microsomes, OKY-046 (greater than 10(-5) M) produced more than 90% inhibition of TXA2 production, whereas platelet aggregation was less inhibited, about 40% inhibition over control, by OKY-046 in that concentration. In the presence of aortic microsomes, the inhibitory effect of OKY-046 on platelet aggregation was markedly augmented in a dose-dependent manner in proportion to the increment of PGI2 production, which paralleled the OKY-046-induced inhibition of TXA2. These results suggest that a selective TXA2 blockade produces effects on platelet aggregation mainly in dual fashion in the presence of PGI2 synthetase: one is due to mere inhibition of TXA2 synthetase and the other is due to the enhancement of PGI2 production probably involving "prostaglandin H2 (PGH2) steal" mechanism, in which PGH2 accumulated in platelets is partly converted to a substrate of PGI2 synthetase in aortic microsomes to produce PGI2.

Topics: Acrylates; Animals; Aorta; Arachidonic Acid; Arachidonic Acids; Cattle; Humans; In Vitro Techniques; Methacrylates; Microsomes; Platelet Aggregation; Prostaglandin Endoperoxides; Prostaglandin Endoperoxides, Synthetic; Prostaglandin H2; Prostaglandins H