Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.5 (thrombin)
 33,306 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We wished to determine whether the metabolism of arachidonic acid, through lipoxygenase and cytochrome P-450 pathways, is involved in production of endothelium-derived relaxing factor(s) (EDRFs) in canine femoral veins. Veins were removed from anesthetized dogs and cut into rings. Endothelium was deliberately removed from some rings. In separate sets of experiments, rings were incubated with either AA861 (10(-5) M) or TMK777 (10(-6) M), inhibitors of 5-lipoxygenase, nordihydroguaiaretic acid (NDGA 3 x 10(-6) M), an inhibitor of lipoxygenase or proadifen (SKF 525A, 10(-6) M), an inhibitor of cytochrome P-450. In addition, some rings were incubated with a combination of indomethacin (10(-5) M) and NG-monomethyl-L-arginine (L-NMMA 10(-4) M) or, where appropriate, a solvent control. Concentration-response curves were obtained for acetylcholine, adenosine diphosphate, thrombin, A23187, and nitric oxide in rings contracted with a submaximal concentration of prostaglandin F2 alpha. AA861 and TMK777 did not alter endothelium-dependent relaxations to the agonists, whether with or without indomethacin and L-NMMA. However, indomethacin plus L-NMMA reduced endothelium-dependent relaxations to thrombin. These results suggest that metabolism of arachidonic acid, through lipoxygenase and cytochrome P-450 pathways, does not produce an EDRF in veins. However, thrombin receptor-activated relaxations are mediated in part by products of the cyclooxygenase pathway and nitric oxide.
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PMID:Role of lipoxygenase and cytochrome P-450 in production of endothelium-derived relaxing factors in canine femoral veins. 127 84

The effect of platelets on polymorphonuclear leukocytes (PMN) O2- production was examined using autologous sheep and human cell systems. Coincubation of sheep platelets with sheep PMNs in the absence of thrombin resulted in a significant inhibition in basal PMN O2- production. The platelet-derived inhibitory activity was released into the medium and could be destroyed by adenosine deaminase suggesting that the inhibitor was adenosine. Addition of alpha-thrombin or platelet activating factor (PAF) enhanced PMN O2- production but only when platelets were present. The enhancement of O2- production in response to thrombin was dependent upon the thrombin concentration and the platelet-PMN ratio. With a platelet: PMN ratio of 30: 1, addition of 10 nM thrombin to sheep cells resulted in a 5-fold increase in O2- production, whereas addition of 10 nM PAF caused a 2-fold increase in O2-. Addition of thrombin or PAF to either PMNs or platelets by themselves did not initiate an increase in O2- generation. The response of human cells was similar except that both thrombin and PAF triggered a 2-fold increase in PMN O2- production in the presence of platelets. The platelet-derived enhancement activity was not released into the medium and was not blocked by WEB 2086, NDGA, ETYA, aspirin or adenosine deaminase. The enhancement effect appeared to be localized to the platelet membrane and we believe requires platelet-PMN contact.
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PMID:Platelet modulation of neutrophil superoxide anion production. 216 Jan 33

Using radioimmunoassay techniques we studied the formation of the 5-lipoxygenase-derived cysteinyl-leukotrienes (LT) in comparison to the cyclooxygenase product thromboxane (TX) B2 in whole human blood allowed to clot at 37 degrees C in vitro. Spontaneous clotting resulted in a time-dependent release of smaller amounts of cysteinyl-LT as well as release of large amounts of TXB2 into the serum. Cysteinyl-LT were characterized by their immunoreactive behaviour and their biological activity in the guinea pig ileum bioassay, an effect which could be antagonized by the SRS-A antagonist FPL 55712 (0.38 microM). By reversed phase HPLC cysteinyl-LT in the serum were identified as a mixture of LTC4, LTD4 and LTE4. At 90 and 120 min part of the immunoreactive material consisted of the omega-oxidized metabolite 20-OH-LTE4. Almost complete inhibition of cyclooxygenase activity by indomethacin (2.8 microM) did not affect cysteinyl-LT formation by clotting whole human blood in vitro nor did activation of platelets by compounds such as the TX mimetic U 46619 (10 microM), platelet-activating factor (PAF, 1 microM) or thrombin (3 IU/ml). In contrast, the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA, 10 microM), the Ca2+-chelating anticoagulants trisodium citrate (10 microM) and edetate disodium (EDTA, 5.4 mM) as well as the functionally unrelated heparin (20 IU/ml) significantly inhibited the formation of cysteinyl-LT as well as of TXB2. Thus, an event related to the process of clotting of whole human blood appears to be able to induce cysteinyl-LT formation in amounts which might be functionally relevant during thromboembolic events.
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PMID:Clotting of whole human blood induces cysteinyl-leukotriene formation. 254 55

4 beta-Phorbol-12-myristate-13-acetate (PMA) at 100 ng/ml was able to induce platelet aggregation in the presence of agents which inhibited aggregation, triggered by other agonists such as adenosine diphosphate sodium salt (ADP), thrombin and collagen. PMA induced aggregation in acid-citrate-dextrose platelet-rich plasma. 100 microM tetracaine, 5 microM bromophenacyl bromide and 0.2 mM mepacrine decreased PMA-induced aggregation by only 10% in contrast to their high inhibitory effect on other aggregation systems. However, 0.4 mM mepacrine did inhibit PMA-induced aggregation at the same rate as the other aggregation systems. 100 mg/ml vincristine slightly affected PMA-induced platelet aggregation, whereas cytochalasin B rather enhanced it. Nordihydroguaiaretic acid, 5, 8, 11, 14-eicosatetraynoic acid and p-nitrophenyl-phosphorylcholine had no effect on PMA- or collagen-induced platelet aggregation, partially inhibited aggregation triggered by ADP and strongly inhibited aggregation caused by thrombin. It is suggested that PMA exerts its effect on platelets mainly due to its ability to alter their membranes.
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PMID:Phorbol-myristate-acetate-induced platelet aggregation in the presence of inhibitors. 314 99

F2-isoprostanes are isomers of the prostaglandin PGF2alpha. At least one compound of this group, 8-epi-PGF2alpha, exhibits biological activity, and therefore special interest is focused on the mechanism of isoprostane formation: enzyme catalyzed or radical mediated. We analyzed the formation of isoprostanes in vitro and in vivo. In both systems, purified cyclooxygenase isoenzymes and cell models specific for the cyclooxygenase isoenzymes, 8-epi-PGF2alpha formation could be totally suppressed by cyclooxygenase inhibitors. Indomethacin inhibited concentration-dependent 8-epi-PGF2alpha formation in platelets stimulated with calcium ionophore, arachidonic acid or thrombin. Nordihydroguaiaretic acid, an antioxidant, blocked isoprostane formation with a similar IC50 value as thromboxane B2 synthesis, pointing toward cyclooxygenase as the primary target of inhibition. Based on the turnover number, cyclooxygenase-2 formed higher levels of 8-epi-PGF2alpha than cyclooxygenase-1. Endogenous 8-epi-PGF2alpha production in rat mesangial cells correlated well with the mRNA and protein expression of cyclooxygenase-2 during interleukin-1 induction. However, in contrast to human platelets, which produced different forms of isoprostanes, rat mesangial cells appeared to form only 8-epi-PGF2alpha. Further, this indicates that mesangial cells may represent a cellular origin for renal 8-epi-PGF2alpha formation. Next, we analyzed the formation of isoprostanes in humans. A direct correlation was observed between indomethacin treatment and the decrease in 8-epi-PGF2alpha and isoprostane levels, but compared with other prostanoids the inhibition was less pronounced. In summary, based on the in vitro studies, a clear cyclooxygenase-dependent formation of isoprostanes, especially 8-epi-PGF2alpha, was observed. However, in vivo additional formation via cyclooxygenase enzyme-independent mechanisms is likely.
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PMID:Generation of the isoprostane 8-epi-prostaglandin F2alpha in vitro and in vivo via the cyclooxygenases. 931 84

Arachidonic acid (AA) liberation and metabolism via cyclo-oxygenase or lipoxygenases may be an important regulatory pathway for mitogenic signalling in human cultured airway smooth muscle (ASM) cells. In cytokine-treated cells, thrombin markedly enhances production of the anti-mitogenic arachidonic acid metabolite, PGE(2). In this study, in the absence of cytokines, we examined the role of endogenous AA metabolism in thrombin-stimulated ASM DNA synthesis. Selective inhibitors of cyclo-oxygenase of 5-lipoxygenase metabolism had no significant effect on 0.3 U/ml thrombin-stimulated DNA synthesis. However, the non-selective, redox-active lipoxygenase inhibitors NDGA and BWA4C inhibited thrombin-stimulated DNA synthesis. Under basal conditions, and following stimulation by thrombin, the levels of the AA metabolites PGE(2), TxA(2), and LTC(4), remained below assay detection limits. Exogenous addition of AA, LTD(4), or 5-, 12-, and 15-HETE and HpETE metabolites had no consistent or substantial stimulatory effect on either basal or thrombin-stimulated DNA synthesis. These data suggest that the non-selective lipoxygenase inhibitors influence DNA synthesis via effects unrelated to lipoxygenase inhibition. The lack of detection of AA metabolites, the lack of influence of selective antagonists/inhibitors of the AA pathway, and the failure of selected AA metabolites to either enhance or directly stimulate DNA synthesis suggest that in the absence of cytokines, cyclo-oxygenase and lipoxygenase metabolism has little role in signalling of human ASM DNA synthesis by thrombin.
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PMID:Thrombin-stimulated DNA synthesis in human cultured airway smooth muscle occurs independently of products of cyclo-oxygenase or 5-lipoxygenase. 1100 67