Cumulative antiplatelet effect of low-dose enteric coated aspirin

Enteric coated aspirin (ECA) at doses of 325-1300 mg is an effective alternative to regular aspirin for inhibition of platelet activity while avoiding gastric irritation. The objectives of this study were to determine: (1) the lowest chronic dose of ECA providing effective inhibition of platelet activities, (2) the time course of the inhibition, and (3) the reappearance of platelet cyclo-oxygenase activity. Seven subjects were studied before and after seven daily doses of 40-325 mg ECA. Serum thromboxane (TX) B2 levels indicated that the lowest dose of ECA resulting in greater than 90% inhibition of platelet cyclo-oxygenase was 80 mg/d. Platelet aggregation and ATP release in response to collagen (1 microgram/ml) and arachidonic acid (1 mM) were abolished and bleeding times were prolonged from 6.1 +/- 1.5 min to 9.7 +/- 2.8 min (mean +/- SD, P less than 0.01). Examination of platelet cyclo-oxygenase activity on a daily basis revealed that 24 h after the first 80 mg dose serum TXB2 had decreased by approximately 60% and was suppressed by more than 90% after four doses. Recovery of platelet cyclo-oxygenase activity after a single 80 mg dose of ECA was delayed for 48-72 h indicating that aspirin reached the systemic circulation. We conclude that chronic inhibition of platelet activity may be achieved in a cumulative manner with 80 mg ECA/d.     

Effects of dipyridamole and low-dose aspirin therapy on platelet adhesion to vascular subendothelium

The effect on platelet function of low-dose aspirin (ASA) and dipyridamole alone or in combination was evaluated after repeated dosing in 5 healthy volunteers. The subjects were treated according to a randomized, single-blind, crossover design with 150 mg of dipyridamole, 25 mg of ASA, the 2 drugs together or placebo twice a day for 3 days. Platelet adhesion was evaluated using an experimental model of adhesion to rat aorta subendothelium under controlled hemodynamic conditions in the presence of red blood cells. Dipyridamole significantly reduced platelet adhesion both alone and in combination with ASA. ASA by itself did not significantly modify platelet adhesion, but completely blocked serum thromboxane production and platelet aggregation by arachidonic acid. Thus, low-dose ASA and dipyridamole may have a complementary action, modifying at the same time 2 platelet functions, adhesion and aggregation, both relevant in the pathogenesis of thrombosis.     

In vivo production of prostacyclin and thromboxane in patients with acute myocardial infarction

The in vivo production of prostacyclin and thromboxane was monitored by measuring their major urinary metabolites 2,3-dinor-thromboxane B2 and 2,3-dinor-6-keto-prostaglandin F1 alpha in ten patients with acute myocardial infarction, five on standard treatment and five receiving prostacyclin infusion. During acute myocardial infarction excretion of 2,3-dinor-thromboxane B2 and 2,3-dinor-6-keto-prostaglandin F1 alpha, measured by a gas chromatography-mass spectrometry method with deuterated internal standards, was significantly increased. This indicates that thromboxane and prostacyclin synthesis are increased during the development of acute myocardial infarction. The excretion data for 2,3-dinor-thromboxane B2 showed that after administration of aspirin there was less pronounced and more variable inhibition than expected. Prostacyclin infusion did not markedly affect the excretion of the thromboxane metabolite.     

In vivo production of prostacyclin and thromboxane in patients with acute myocardial infarction.

The in vivo production of prostacyclin and thromboxane was monitored by measuring their major urinary metabolites 2,3-dinor-thromboxane B2 and 2,3-dinor-6-keto-prostaglandin F1 alpha in ten patients with acute myocardial infarction, five on standard treatment and five receiving prostacyclin infusion. During acute myocardial infarction excretion of 2,3-dinor-thromboxane B2 and 2,3-dinor-6-keto-prostaglandin F1 alpha, measured by a gas chromatography-mass spectrometry method with deuterated internal standards, was significantly increased. This indicates that thromboxane and prostacyclin synthesis are increased during the development of acute myocardial infarction. The excretion data for 2,3-dinor-thromboxane B2 showed that after administration of aspirin there was less pronounced and more variable inhibition than expected. Prostacyclin infusion did not markedly affect the excretion of the thromboxane metabolite.     

Increased prostacyclin and thromboxane A2 biosynthesis in atherosclerosis.

It has been proposed that atherosclerotic arteries produce less prostacyclin (PGI2) than nonatherosclerotic arteries do, thereby predisposing arteries to vasospasm and thrombosis in vivo. We reexamined this concept by measuring spontaneous as well as arachidonate-induced PGI2 biosynthesis in aortic segments from nonatherosclerotic and cholesterol-fed atherosclerotic New Zealand White rabbits. Thromboxane A2 (TXA2) generation was also measured. Formation of PGI2, as well as TXA2, as measured by radioimmunoassay (RIA) of their metabolites, was increased in atherosclerotic aortic segments relative to nonatherosclerotic segments (P less than or equal to 0.05) at 0, 5, 10, 15, and 30 min of incubation with arachidonate. Pretreatment of arterial segments with indomethacin inhibited PGI2 as well as TXA2 formation, whereas pretreatment with the selective TXA2 inhibitor OKY-046 inhibited only TXA2 release, thus confirming the identity of icosanoids. To confirm the RIA data, aortic segments were incubated with [14C]arachidonate prior to stimulation with unlabeled arachidonate. The uptake of arachidonate was similar, but the release of incorporated [14C]arachidonate was significantly (P less than or equal to 0.05) greater in atherosclerotic segments than in nonatherosclerotic ones. Conversions of released [14C]arachidonate to 6-keto[14C]prostaglandin F1 alpha and [14C]thromboxane B2 were similar in the two types of aortic segments. Thus, synthesis of PGI2 as well as TXA2 is increased in atherosclerosis, and this alteration in arachidonate metabolism is related to increased release of arachidonate.     

Inhibition of prostacyclin and thromboxane A2 generation by low-dose aspirin at the site of plug formation in man in vivo

In a double-blind placebo-controlled crossover study, we investigated in seven healthy male volunteers the effect of a low-dose aspirin regimen (35 mg acetylsalicylate per day for 7 days) on the formation of thromboxane A2 (TxA2) and prostacyclin (PGI2) in blood emerging from a standardized injury of the microvasculature made to determine skin bleeding time. When subjects were treated with placebo, there was rapid and substantial generation of TxA2 and PGI2 at the site of platelet-vessel wall interaction within the first 2 min after vascular injury. This was reflected by a greater than 100-fold and greater than 10-fold increase in thromboxane B2 (TxB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) in blood obtained from incisions made to determine bleeding time as compared with the corresponding plasma values. Low-dose aspirin caused a significant inhibition of both TxA2 and PGI2 generation in blood sampled from the skin incisions, represented by a 85% and 92% and 81% and 84% inhibition of TxB2 and 6-keto-PGF1 alpha, respectively, as compared with controls. We therefore conclude that rapid activation of both platelet prostaglandin metabolism and vascular PGI2 biosynthesis occurs at the site of platelet-vessel wall interaction, and low-dose aspirin results in a significant inhibition of both platelet and vascular cyclooxygenase activity. Thus, our data fail to confirm the concept of a differential effect of low-dose aspirin on platelet and vascular prostaglandin synthesis in man in vivo.     

Intestinal prostacyclin and thromboxane production in irreversible hemorrhagic shock

Arterial and intestinal venous blood were sampled every hour for measurement of thromboxane B2 (TXB2) and 6-keto-PGF1 alpha, stable metabolites of thromboxane A2 and prostacyclin, respectively, in dogs subjected to hemorrhagic hypotension at 32.8 +/- 1.4 mm Hg for 3 h, followed by reinfusion of the remaining shed blood. Control dogs were treated alike without hypotension. Arterial and intestinal venous TXB2 significantly increased during hypotensive and post-transfusion periods, the venous concentration being significantly higher than the corresponding arterial. The arterial and venous 6-keto-PGF1 alpha increased during hypotension but decreased during post-transfusion periods. Furthermore, arterial and venous TXB2 to 6-keto-PGF1 alpha concentration ratio increased. Intestinal TXB2 release (blood flow X arteriovenous concentration difference) increased progressively, whereas 6-keto-PGF1 alpha release decreased. No significant changes occurred in the control dogs. This study shows an imbalance in intestinal production and release of TXA2 and PGI2, in favor of TXA2 during severe hemorrhagic hypotension and after blood transfusion. The imbalance may contribute to the development of irreversible hemorrhagic shock and reperfusion injury.     

小剂量阿司匹林对老年缺血性脑血管病患者血小板聚集功能的影响

目的；观察小剂量阿司匹林（ASA）对老年缺血脑血管病（ICVD）患者血小板聚集功能的影响。方法：以花生四烯酸，二磷酸腺苷，肾上腺素和胶原为诱导剂检测了老年ICVD患者服用ASA（40mg/d)70例，未服ASA30例及健康对照组50例的血小板最大聚集率（MAR）。结果：40mg/d的ASA组中花生四烯酸，胶原和肾上腺素诱导MAR的变异系数较大，且70例中有25例对花生四烯酸诱导的聚集无显著抑制。此25例中随机选取10例ASA增至80mg/d后，有8例达到了对花生四烯酸聚集的显著抑制，另2例继续增至100mg/d后才达到。结论：在老年ICVD患者中，小剂量ASA作用的个体差异较大，临床ASA效果需要实验室评价并需个体化。     

Endogenous biosynthesis of thromboxane and prostacyclin in 2 distinct murine models of atherosclerosis

Thromboxane A(2) is a potent vasoconstrictor and platelet agonist; prostacyclin is a potent platelet inhibitor and vasodilator. Altered biosynthesis of these eicosanoids is a feature of human hypercholesterolemia and atherosclerosis. This study examined whether in 2 murine models of atherosclerosis their levels are increased and correlated with the evolution of the disease. Urinary 2,3-dinor thromboxane B(2) and 2,3-dinor-6-keto prostaglandin F(1 alpha), metabolites of thromboxane and prostacyclin, respectively, were assayed in apoliprotein E (apoE)-deficient mice on chow and low-density lipoprotein receptor (LDLR)-deficient mice on chow and a Western-type diet. Atherosclerosis lesion area was measured by en face method. Both eicosanoids increased in apoE-deficient mice on chow and in LDLR-deficient mice on a high-fat diet, but not in LDLR-deficient mice on chow by the end of the study. Aspirin suppressed ex vivo platelet aggregation, serum thromboxane B(2), and 2,3-dinor thromboxane B(2), and significantly reduced the excretion of 2,3-dinor-6-keto prostaglandin F(1 alpha) in these animals. This study demonstrates that thromboxane as well as prostacyclin biosynthesis is increased in 2 murine models of atherogenesis and is secondary to increased in vivo platelet activation. Assessment of their generation in these models may afford the basis for future studies on the functional role of these eicosanoids in the evolution and progression of atherosclerosis. (Blood. 2000;96:3823-3826)     

Selective inhibition of platelet cyclooxygenase with controlled release, low-dose aspirin

The hypothesis that slow administration of low doses of aspirin may selectively inhibit platelet cyclooxygenase and thromboxane A₂ formation was evaluated using controlled release aspirin formulations. In the first study, doses of either 50, 100, 325 and 1,300 mg of these formulations and 300 mg soluble aspirin were ingested daily by healthy volunteers for one week. In the second study, doses of 5, 10, 25 and 50 mg controlled release aspirin, 50 mg soluble aspirin and 100 mg aspirin and glycine formulation were ingested daily for ten days. Platelet function and urinary prostaglandin production were assessed immediately before and on the seventh day of dosing in both studies and in the second study, repeated on the tenth day of dosing. Platelet function and serum thromboxane B₂ production were fully inhibited by all formulations of 50 mg aspirin and above, but not by doses of controlled release aspirin below 50 mg doses. The excretion of urinary 6-keto-PGF₁α (a major metabolite of prostacyclin) was significantly reduced at controlled release aspirin doses above 100 mg and at all doses of rapidly absorbed aspirin tested. As no significant reduction in the urinary 6-keto-PGF₁α production was observed at doses of controlled release aspirin of 50 and 100 mg and below, it appeared that these doses did not inhibit the systemic vascular cyclooxygenase. These data are consistent with a selective inhibition of platelet function by daily doses of 50 and 100 mg of the controlled release formulation of aspirin.     

Endotoxin-induced production of thromboxane and prostacyclin by equine peritoneal macrophages

Equine peritoneal macrophages were isolated and cultured in vitro to assess their ability to produce thromboxane (TxA2) and prostacyclin (PGI2) in response to endotoxin. Peritoneal macrophages (2.5 x 10(6)/ml) were incubated in tissue culture media, containing 1) no additive (nonstimulated control), 2) endotoxin (0.5 to 100 ng/ml) or 3) the calcium ionophore, A23187 (0.95 microM) for two and six h. Concentrations of the stable metabolites of TxA2 and PGI2 thromboxane B2 (TxB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), in the incubation media were determined by radioimmunoassay. The concentrations of both metabolites increased from two to six h incubation. Endotoxin increased the production of TxA2 and PGI2 over the nonstimulated control values at both two and six h and endotoxin-induced concentrations of 6-keto-PGF1 alpha were higher at six than at two h. The response of macrophages to A23187 was similar to endotoxin. Mean eicosanoid concentrations did not differ among the range of endotoxin concentrations at either time; however there was significant curvilinear regression between endotoxin concentration and TxB2 at both times, and between endotoxin and 6-keto-PGF1 alpha at two h. The results indicate that equine macrophages may be a significant source of TxA2 and PGI2 during endotoxemia.     

The effect of aspirin on mean platelet volume in patients with paroxysmal atrial fibrillation

Aspirin is one of the preferred therapies in the primary prevention of ischemic stroke in paroxysmal atrial fibrillation (PAF). Mean platelet volume (MPV) is a marker of platelet size and activation. Increased MPV reflects active and large platelets. The present observational study was designed to investigate whether aspirin treatment does affect MPV levels in patients with PAF. The study included 101 patients who were detected to have PAF by 24-hour Holter monitoring and divided into two groups based on aspirin treatment [ASA (+) and ASA (?)]. MPV was measured. Patients with aortic and mitral stenosis, hyperthyroidism, hypothyroidism, malignancy, infection, and pregnancy were excluded from the study. Of the 101 patients, 50 had no antiplatelet therapy and 51 had daily aspirin (100?mg) intake. Mean age of the patients was 66?±?10 years and 35 (68%) were male in ASA (+) group. There was no difference in median levels of MPV (9.9 vs. 10.2?fl, respectively; p?=?0.9) between groups. Both uni- and multivariate logistic regression analyses did not show an association between MPV and ASA use. Our results indicate that MPV as a predictive marker of platelet size and activity is not affected by aspirin use in patients with PAF.     

Imbalance of thromboxane/prostacyclin biosynthesis in patients with lupus anticoagulant

The mechanism involved in the association between antiphospholipid antibodies and thrombosis or fetal loss remains unclear. We assessed the biosynthesis of thromboxane A2 and prostacyclin in 31 samples from 25 patients with lupus anticoagulant and in 32 controls. The urinary excretion of the major thromboxane metabolite of platelet origin (11-dehydrothromboxane B2) was very significantly increased (P less than .0003) in the patients. In contrast, the urinary metabolite reflecting the vascular production of prostacyclin (2,3-dinor-6-keto-prostaglandin F1 alpha) was much less increased (P less than .02). We found no correlation between the levels of anticardiolipin antibodies and the urinary excretion of 11-dehydro-thromboxane B2. Six patients with elevated urinary 11-dehydrothromboxane B2 were treated with low-dose aspirin (20 mg/d during 7 days). In these patients, there was a close relationship between the extent of inhibition of the thromboxane urinary metabolite (72%) and serum thromboxane B2 (79%). In contrast, the urinary excretion of 2,3-dinor-6-ketoprostaglandin F1 alpha was nearly unchanged (13% reduction). In addition, the F(ab')2 fragments isolated from six patients presenting increased urinary 11-dehydro-thromboxane B2 enhanced the generation of thromboxane B2 (P = .04) and the release of 14C serotonin (P = .009) by normal washed platelets, as compared with F(ab')2 from controls. In summary, our study shows that in patients with lupus anticoagulant, platelet activation may occur without a compensatory increment in the vascular biosynthesis of prostacyclin. This observation may be crucial to cause or reflect an increased risk for thrombosis. In addition, our results may suggest a rationale for antiplatelet agents for the prophylaxis of thrombosis in many patients with the antiphospholipid syndrome.     

Acute antithrombotic effect of a front-loaded regimen of clopidogrel in patients with atherosclerosis on aspirin

There is a need for a rapid antithrombotic effect after the administration of antiplatelet drugs in the setting of acute coronary syndromes and percutaneous interventions. Clopidogrel, a new thienopyridine derivative, is an efficient antiplatelet agent. However, the standard regimen of clopidogrel (75 mg/d) requires 2 to 3 days before significant antithrombotic effects. Patients with stable arterial disease on chronic aspirin therapy (n=20) were treated with clopidogrel either with a front-loaded regimen, 300 mg the first day and 75 mg/d the next 7 days, or with a standard regimen, 75 mg/d for 8 days. Blood thrombogenicity was assessed by quantification of platelet-thrombus formation in an ex vivo perfusion chamber, by ADP-induced platelet aggregation, and by ADP-induced fibrinogen binding. At 2 hours, mean total thrombus area with the standard regimen was not significantly reduced. In contrast, at 2 hours, the mean total thrombus area with the front-loaded regimen was significantly decreased by 23.1+/-8.5% versus baseline (P<0.05). ADP-induced platelet aggregation (with 5 and 10 micromol/L) was also significantly (P<0.05) reduced with the front-loaded regimen at 2 hours, with the mean platelet aggregation being 82.2+/-4.4% and 81.8+/-4.5%, respectively, versus baseline. Similarly, flow cytometry demonstrated a significant decrease (P<0. 05) in the ADP-induced fibrinogen binding (with 0.12 and 0.6 micromol/L) at 2 hours in this front-loaded regimen group (36.1+/-2. 0% and 53.2+/-9.3%). With the standard regimen, platelet activity was not significantly reduced at 2 hours. Our data suggest that a front-loaded regimen of clopidogrel added to aspirin achieves a significant antithrombotic effect at 2 hours in patients with known atherosclerotic disease on chronic aspirin therapy. This provides a rationale for using front-loaded clopidogrel in combination with aspirin in percutaneous coronary interventions.     

An inhibitory effect of camonagrel-a new thromboxane synthase inhibitor, on P-selectin-mediated platelet/PMN adhesion

P-selectin (PADGEM protein, GMP-140 or CD 62) is a glycoprotein of platelet a-granules and endothelial Weibel-Palade bodies that, by mediating cellular adhesion, initiates recruitment of leukocytes and lymphocytes into injured tissue. Both of the endothelial antiplatelet autacoids prostacyclin (PGI(2)) and nitric oxide (NO) have been demonstrated to inhibit P-selectin expression. Prostaglandin endoperoxides PGG(2)/PGH(2) that are generated by activated platelets have been demonstrated to be used by endothelium for generation of prostacyclin. In an experimental model in vitro that resembles vessel wall/platelet/PMN interaction in vivo, we found that aspirin (100 μM), a COX inhibitor, but not L-NMMA (100 μM) and a NO-synthase inhibitor, reversed the inhibitory effect of arterial wall on P-selectin mediated platelet/PMN adhesion. The anti-adhesive potency of vessel wall reversed by aspirin was dose-dependently restored by camonagrel (3-100 μM), a new TXA(2) synthase inhibitor. We conclude that selective TXA(2)-synthase inhibitors may inhibit P-selectin mediated platelet/PMN adhesion by augmenting formation of prostacyclin by vessel walls.     

The inhibitory effect of PGI2 (prostacyclin) on white platelet arterial thrombus formation.

Prostaglandins play an important role in the platelet-vessel wall interactions. Superfusion with PGI2 of arterial segments prepared in an in vivo model for thrombus induction resulted in a very marked dose-effect-related decrease of white arterial thrombus formation.     

Differential inhibitory actions by glucocorticoid and aspirin on cytokine-induced nitric oxide production in vascular smooth muscle cells

Glucocorticoids and nonsteroidal antiinflammatory drugs (NSAIDs) are widely used for the treatment of inflammatory and immune diseases. Nitric oxide (NO) has a diversity of physiological functions, but its excess production has been implicated in the inflammatory process. The present study was designed to elucidate the mechanisms by which glucocorticoids and NSAIDs affect inducible nitric oxide synthase (iNOS) expression in cultured rat vascular smooth muscle cells (VSMCs). Both interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha potently stimulated nitrite/nitrate (NOx) production with a concomitant expression of iNOS mRNA and protein as demonstrated by Northern and Western blot analysis, respectively. Both IL-1beta and TNF-alpha activated nuclear factor (NF)-kappaB as demonstrated by electrophoretic mobility shift assay. Dexamethasone, salicylate and aspirin, but not indomethacin, dose dependently inhibited cytokine-stimulated NOx production and iNOS protein expression. Dexamethasone decreased cytokine-induced NF-kappaB activation and iNOS mRNA expression, but neither salicylate nor aspirin affected NF-kappaB activation or iNOS mRNA expression. IL-1beta caused a rapid increase in phosphorylated IkappaB-alpha levels and subsequent transient decrease in IkappaB-alpha levels, an inhibitor of NF-kappaB, as revealed by Western blot analysis using specific antibodies for phosphorylated and nonphosphorylated IkappaB-alpha. These effects were blocked by pretreatment with dexamethasone. Aspirin dose dependently inhibited iNOS enzymatic activity, whereas salicylate and dexamethasone had limited effect. The present study demonstrates that 1) inhibitory effect of dexamethasone on cytokine-induced iNOS expression and NO production in rat VSMCs, although potentially acting at multiple levels, is partly mediated by inhibition of NF-kappaB activation resulting from decreased phosphorylation and degradation of IkappaB-alpha, 2) both salicylate and aspirin inhibit cytokine-stimulated NO production at translational and/or posttranslational levels without affecting NF-kappaB- mediated iNOS gene expression, and 3) aspirin directly inhibits iNOS enzyme activity. These data suggest the differential inhibitory mechanisms of iNOS-mediated NO synthesis by glucocorticoids and NSAIDs in the vasculature.     

Gastric changes in coronary-operated patients with low-dose aspirin.

Low-dose aspirin is widely used in patients operated on for coronary disease as secondary prevention of coronary artery occlusion. The changes caused by aspirin in therapeutic doses to gastric mucosa are well documented, but the effect of long-term low-dose aspirin is not so well known. Forty-six volunteer coronary-operated patients with daily low-dose aspirin were interviewed postoperatively, and an upper gastrointestinal tract endoscopy was performed and biopsy specimens taken 3 months after the operation. The findings were compared with a normal population sample of 358 persons from a study previously published. There were significantly more erosions and ulcers or fresh scars in the study group than in the control population--11 of 46 patients and 24 of 358 patients, respectively. The presence of superficial gastritis was similar. Mostly, the lesions were asymptomatic. History of peptic ulcer disease, use of other ulcerogenic drugs, smoking, and alcohol consumption had no predictive value for acute lesions. In contrast, the lesions were associated with chronic superficial gastritis and Helicobacter pylori infection.