RESUMO
RATIONALE: The balance between vascular prostacyclin, which is antithrombotic, and platelet thromboxane A2, which is prothrombotic, is fundamental to cardiovascular health. Prostacyclin and thromboxane A2 are formed after the concerted actions of cPLA2α (cytosolic phospholipase A2) and COX (cyclooxygenase). Urinary 2,3-dinor-6-keto-PGF1α (PGI-M) and 11-dehydro-TXB2 (TX-M) have been taken as biomarkers of prostacyclin and thromboxane A2 formation within the circulation and used to explain COX biology and patient phenotypes, despite concerns that urinary PGI-M and TX-M originate in the kidney. OBJECTIVE: We report data from a remarkable patient carrying an extremely rare genetic mutation in cPLA2α, causing almost complete loss of prostacyclin and thromboxane A2, who was transplanted with a normal kidney resulting in an experimental scenario of whole-body cPLA2α knockout, kidney-specific knockin. By studying this patient, we can determine definitively the contribution of the kidney to the productions of PGI-M and TX-M and test their validity as markers of prostacyclin and thromboxane A2 in the circulation. METHODS AND RESULTS: Metabolites were measured using liquid chromatography-tandem mass spectrometry. Endothelial cells were grown from blood progenitors. Before kidney transplantation, the patient's endothelial cells and platelets released negligible levels of prostacyclin (measured as 6-keto-prostaglandin F1α) and thromboxane A2 (measured as TXB2), respectively. Likewise, the urinary levels of PGI-M and TX-M were very low. After transplantation and the establishment of normal renal function, the levels of PGI-M and TX-M in the patient's urine rose to within normal ranges, whereas endothelial production of prostacyclin and platelet production of thromboxane A2 remained negligible. CONCLUSIONS: These data show that PGI-M and TX-M can be derived exclusively from the kidney without contribution from prostacyclin made by endothelial cells or thromboxane A2 by platelets in the general circulation. Previous work relying on urinary metabolites of prostacyclin and thromboxane A2 as markers of whole-body endothelial and platelet function now requires reevaluation.
Assuntos
6-Cetoprostaglandina F1 alfa/análogos & derivados , Aloenxertos/metabolismo , Transplante de Rim , Rim/metabolismo , Mutação com Perda de Função , Fosfolipases A2 Citosólicas/genética , Tromboxano B2/análogos & derivados , 6-Cetoprostaglandina F1 alfa/metabolismo , 6-Cetoprostaglandina F1 alfa/urina , Biomarcadores/urina , Células Cultivadas , Feminino , Humanos , Pessoa de Meia-Idade , Fenótipo , Fosfolipases A2 Citosólicas/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Tromboxano B2/metabolismo , Tromboxano B2/urinaRESUMO
Established "low" aspirin dosages inconsistently inhibit platelet function in dogs. Higher aspirin dosages consistently inhibit platelet function, but are associated with adverse effects. The objectives of this study were to use an escalation in dosage to determine the lowest aspirin dosage that consistently inhibited platelet function without inhibiting prostacyclin synthesis. Eight dogs were treated with five aspirin dosages: 0.5 mg/kg q24h, 1 mg/kg q24h, 2 mg/kg q24h, 4 mg/kg q24h and 10 mg/kg q12h for 7 days. Utilizing aggregometry and a whole-blood platelet function analyzer (PFA-100), platelet function was evaluated before and after treatment. Urine 11-dehydro-thromboxane-B2 (11-dTXB2 ) and 6-keto-prostaglandin-F1α (6-keto-PGF1α ), were measured. Compared to pretreatment, there were significant post-treatment decreases in the maximum aggregometry amplitude and increases in the PFA-100 closure times for all dosages expect 0.5 mg/kg q24h. There was no difference in amplitude or closure time among the 2 mg/kg q24h, 4 mg/kg q24h, and 10 mg/kg q12h dosages. Compared to pretreatment values, there was a significant decrease in urinary 11-dTXB2 -to-creatinine and 6-keto-PGF1α -to-creatinine ratios, but there was no dose-dependent decrease for either metabolite. An aspirin dosage of 2 mg/kg q24h consistently inhibits platelet function without decreasing prostacyclin synthesis significantly more than lower aspirin dosages.
Assuntos
Aspirina/farmacologia , Plaquetas/efeitos dos fármacos , Epoprostenol/urina , Tromboxanos/urina , 6-Cetoprostaglandina F1 alfa/urina , Animais , Aspirina/administração & dosagem , Cães , Relação Dose-Resposta a Droga , Feminino , Masculino , Agregação Plaquetária/efeitos dos fármacos , Testes de Função Plaquetária/veterinária , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
We hypothesise that molecules in the cyclooxygenase pathway affect platelet activity when seminal fluid (SF) is present. We considered the influence of SF on platelet aggregation in women, and believe that the prostanoids in SF signalling are significant. Thirty-one female subjects were studied, 20 of whom were sexually active. Male partners were given either aspirin or indomethacin to inhibit cyclooxygenase. The 6-keto prostaglandin F1α (6-keto PGF1α) and prostaglandin E metabolite (PGE-M) in SF were measured by competitive assay. Platelets and prostanoids were evaluated in women, periodically, before and after intercourse. The platelets were tested with adenosine diphosphate (ADP) and arachidonic acid (AA). To block the interaction between the uterus and SF, some couples used condoms. We found that the 6-keto prostaglandin F1α in urine at 2 hours post-intercourse (1418.75 pg/mL, Std 688.39) was greater than pre-intercourse (772.68 pg/mL, Std 116.54). Post-intercourse, a transient decrease in platelet aggregation was observed in women whose partners did not use condoms. Averages for platelet aggregation were 20.16% with ADP, and more significantly, 37.79% with AA after 2 hours. In contrast, couples using condoms showed no changes, averaging 64.02% with ADP and 72.06% with AA. Women whose partners were taking aspirin or indomethacin also showed no changes. SF from men taking aspirin or indomethacin led to no reduction in platelet aggregometry in their partners. These results indicate that in cases of exposure to SF, the transient change in women's platelet activity could be related to the cyclooxygenase pathway.
Assuntos
Coito , Agregação Plaquetária , Prostaglandina-Endoperóxido Sintases/metabolismo , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Alprostadil/análogos & derivados , Alprostadil/urina , Aspirina/farmacologia , Preservativos , Feminino , Humanos , Indometacina/farmacologia , Masculino , Pessoa de Meia-Idade , Agregação Plaquetária/efeitos dos fármacos , Sêmen/efeitos dos fármacos , Sêmen/metabolismoRESUMO
BACKGROUND: Acute kidney injury (AKI) is frequently observed in critically ill patients in the intensive care unit (ICU) and is associated with increased mortality. Prostanoids regulate numerous biological functions, including hemodynamics and renal tubular transport. We herein investigated the ability of urinary prostanoid metabolites to predict the onset of AKI in critically ill adult patients. METHODS: The current study was conducted as a prospective observational study. Urine of patients admitted to the ICU at Okayama University Hospital was collected and the urinary levels of prostaglandin E2 (PGE2), PGI2 metabolite (2,3-dinor-6-OXO-PGF1α), thromboxane A2 (TXA2) metabolite (11-dehydro-TXB2) were determined. RESULTS: Of the 93 patients, 24 developed AKI (AKIN criteria). Surgical intervention (93, 75 %) was the leading cause of ICU admission. Overall, the ratio of the level of serum Cr on Day 1 after ICU admission to that observed at baseline positively correlated with the urinary 2,3-dinor-6-OXO-PGF1α/Cr (r = 0.57, p < 0.0001) and 11-dehydro-TXB2/Cr (r = 0.47, p < 0.0001) ratios. In 16 cases of de novo AKI, the urinary 2,3-dinor-6-OXO-PGF1α/Cr and 11-dehydro-TXB2/Cr values were significantly elevated compared with that observed in the non-AKI group, whereas the urinary PGE2/Cr values were not. The urinary 2,3-dinor-6-OXO-PGF1α/Cr ratio exhibited the best diagnostic and predictive performance among the prostanoid metabolites according to the receiver operating characteristic (ROC) analysis [ROC-area under the curve (AUC): 0.75]. CONCLUSIONS: Taken together, these results demonstrate that the urinary 2,3-dinor-6-OXO-PGF1α/Cr and 11-dehydro-TXB2/Cr ratios are associated with the subsequent onset of AKI and poor outcomes in adult heterogeneous ICU patients.
Assuntos
Injúria Renal Aguda/diagnóstico , Injúria Renal Aguda/urina , Prostaglandinas/urina , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Biomarcadores/urina , Creatinina/sangue , Cuidados Críticos , Estado Terminal , Feminino , Humanos , Japão , Masculino , Complicações Pós-Operatórias/terapia , Complicações Pós-Operatórias/urina , Valor Preditivo dos Testes , Estudos Prospectivos , Tromboxano B2/análogos & derivados , Tromboxano B2/urina , Resultado do TratamentoRESUMO
AIM: A genetic variant (rs20417) of the PTGS2 gene, encoding for COX-2, has been associated with decreased COX-2 activity and a decreased risk of cardiovascular disease (CVD). However, this genetic association and the role of COX-2 in CVD remain controversial. METHODS AND RESULTS: The association of rs20417 with CVD was prospectively explored in 49 232 subjects (ACTIVE-A, CURE, epiDREAM/DREAM, ONTARGET, RE-LY, and WGHS) and the effect of potentially modifiable risk factors on the genetic association was further explored in 9363 INTERHEART participants. The effect of rs20417 on urinary thromboxane and prostacyclin metabolite concentrations was measured in 117 healthy individuals. Carriage of the rs20417 minor allele was associated with a decreased risk of major CVD outcomes (OR = 0.78, 95% CI: 0.70-0.87; P = 1.2 × 10(-5)). The genetic effect was significantly stronger in aspirin users (OR: 0.74, 95% CI: 0.64-0.84; P = 1.20 × 10(-5)) than non-users (OR: 0.87, 95% CI: 0.72-1.06; P = 0.16) (interaction P-value: 0.0041). Among patients with previous coronary artery disease (CAD), rs20417 carriers had a stronger protective effect on risk of major adverse events when compared with individuals without previous CAD (interaction P-value: 0.015). Carriers had significantly lower urinary levels of thromboxane (P = 0.01) and prostacyclin (P = 0.01) metabolites when compared with non-carriers. CONCLUSION: The rs20417 polymorphism is associated with a reduced risk of major cardiovascular events and lower levels of thromboxane and prostacyclin. Our results suggest that a genetic decrease in COX-2 activity may be beneficial with respect to CVD risk, especially, in higher risk patients on aspirin.
Assuntos
Ciclo-Oxigenase 2/genética , Infarto do Miocárdio/genética , Polimorfismo Genético/genética , Acidente Vascular Cerebral/genética , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Idoso , Aspirina/uso terapêutico , Inibidores de Ciclo-Oxigenase/uso terapêutico , Feminino , Heterozigoto , Humanos , Masculino , Estudos Multicêntricos como Assunto , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/urina , Estudos Prospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto , Acidente Vascular Cerebral/tratamento farmacológico , Acidente Vascular Cerebral/urina , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
OBJECTIVE: We examined the value of inflammatory and oxidative biomarkers in predicting acute kidney injury (AKI) following orthotopic liver transplantation (OLT). METHODS: Urinary excretion of tumour necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-10 (IL-10), superoxide dismutase (SOD), malondialdehyde (MDA), 6-keto prostaglandin F1α (6-keto-PGF1α), hydrogen peroxide (H2O2), and 8-keto prostaglandin F2α (8-iso-PGF2α), serum creatinine (SCr), blood urea nitrogen (BUN), urinary N-acetyl-beta-D-glucosaminidase (NAG), ß2-microglobulin (ß2-MG) and γ-glutamyl-transferase (γ-GT), were measured before surgery (baseline), at 2 h after graft reperfusion and 24 h after OLT in 28 liver transplantation patients. RESULTS: The levels of TNF-α, IL-8, IL-10, SOD, MDA, 6-keto-PGF1α, H2O2 and 8-iso-PGF2α in urine were all significantly higher in patients who had AKI than in those who did not at 2 h after graft reperfusion and 24 h after OLT (p < 0.01).
Assuntos
Injúria Renal Aguda/diagnóstico , Injúria Renal Aguda/urina , Biomarcadores/urina , Transplante de Fígado , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Dinoprosta/análogos & derivados , Dinoprosta/urina , Feminino , Humanos , Peróxido de Hidrogênio/urina , Interleucina-10/urina , Interleucina-8/urina , Masculino , Malondialdeído/urina , Pessoa de Meia-Idade , Período Pós-Operatório , Período Pré-Operatório , Prognóstico , Curva ROC , Superóxido Dismutase/urina , Fatores de Tempo , Fator de Necrose Tumoral alfa/urinaRESUMO
BACKGROUND: To examine the effect of maternal smoking in pregnancy on the production of two eicosanoids, thromboxane A(2) and prostacyclin I2, and their role in the pathogenesis of intrauterine growth restriction. METHODS: Prospective case control study enrolled smoking and non-smoking women at ≤14 weeks gestation. Maternal urine samples were obtained at ≤14, 28 and 36 weeks. High performance liquid chromatography tandem mass spectrometry (LC-MS-MS) was used to quantify 11-dehydrothromboxane B(2) (TX-M) and 2,3 dinor-6-ketoprostaglandin F1α (PG-M), stable urinary metabolites of thromboxane A(2) and prostacyclin I2. Confirmation of the smoking status was performed by quantitation of urinary nicotine metabolites. Data was analysed using SPSS and Stata(®). RESULTS: Thirty five were enrolled in the smoking group and 32 in the non-smoking group. Smoking resulted higher levels of TX-M at ≤14, 28 and 36 weeks gestation. There was no difference in PG-M at any gestational time point between the two groups. The median customised birthweight centile in the smoking group was 17.0 (0-78) compared to 55.5 (4-100) in the non-smoking group (P<0.001). A causal relationship between elevated TX-M and IUGR could not be established. CONCLUSIONS: Maternal smoking in pregnancy is associated with altered eicosanoid production in favour of the vasoconstrictor thromboxane A(2) which occurs early in the first trimester.
Assuntos
Retardo do Crescimento Fetal/etiologia , Fumar/efeitos adversos , Tromboxano A2/metabolismo , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Estudos de Casos e Controles , Feminino , Retardo do Crescimento Fetal/metabolismo , Humanos , Gravidez , Tromboxano B2/análogos & derivados , Tromboxano B2/urina , Adulto JovemRESUMO
Patients with Down syndrome (DS) and a left-to-right shunt often develop early severe pulmonary hypertension (PH) and pulmonary vascular obstructive disease (PVOD); the pathophysiological mechanisms underlying the development of these complications are yet to be determined. To investigate the mechanisms, we evaluated the biosynthesis of thromboxane (TX) A(2) and prostacyclin (PGI(2)) in four groups of infants, cross-classified as shown below, by measuring the urinary excretion levels of 11-dehydro-TXB(2) and 2,3-dinor-6-keto-PGF(1alpha): DS infants with a left-to-right shunt and PH (D-PH, n = 18), DS infants without congenital heart defect (D-C, n = 8), non-DS infants with a left-to-right shunt and PH (ND-PH, n = 12), and non-DS infants without congenital heart defect (ND-C, n = 22). The urinary excretion ratios of 11-dehydro-TXB(2) to 2,3-dinor-6-keto-PGF(1alpha) in the D-PH, D-C, ND-PH, and ND-C groups were 7.69, 4.71, 2.10, and 2.27, respectively. The ratio of 11-dehydro-TXB(2) to 2,3-dinor-6-keto-PGF(1alpha) was higher in the presence of DS (P < 0.001), independently of the presence of PH (P = 0.297). The predominant biosynthesis of TXA(2) over PGI(2), leading to vasoconstriction, was observed in DS infants, irrespective of the presence/absence of PH. This imbalance in the biosynthesis of vasoactive eicosanoids may account for the rapid progression of PVOD in DS infants with a left-to-right shunt.
Assuntos
Síndrome de Down/metabolismo , Epoprostenol/biossíntese , Cardiopatias Congênitas/metabolismo , Hipertensão Pulmonar/metabolismo , Pneumopatias Obstrutivas/metabolismo , Doença Cardiopulmonar/metabolismo , Tromboxano A2/biossíntese , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Pré-Escolar , Estudos Transversais , Síndrome de Down/complicações , Feminino , Cardiopatias Congênitas/etiologia , Humanos , Hipertensão Pulmonar/etiologia , Lactente , Pneumopatias Obstrutivas/etiologia , Masculino , Prognóstico , Doença Cardiopulmonar/etiologia , Radioimunoensaio , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
Eicosanoids play an important role in the evaluation of pro-inflammatory responses and in the safety and toxicity of novel therapeutic agents. This work describes a high-throughput UFLCMS/MS method for the analysis of three urinary prostanoid biomarkers of pro-inflammatory responses, tetranor PGEm, 6-keto PGF(1alpha) and 2,3-dinor-6-keto PFG(1alpha). Nine male volunteers of various age and fitness level participated in this study. Six provided pre- and post-exercise samples and three provided intraday samples. Tetranor PGEm and 6-keto PGF(1alpha) increased significantly in patients after exercise (p<0.017 and p<0.029). In individual patient sets, tetranor PGEm levels increased from 1.5- to 6-fold pre- vs. post-exercise, levels of 6-keto PGF(1alpha) increased more dramatically from 2- to 55-fold pre- vs. post-exercise. The prostanoid 2,3-dinor-6-keto PGF(1alpha) remained unchanged post-exercise. Data was normalized to urinary creatinine concentration, which increased approximately 40% post-exercise.
Assuntos
6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Cromatografia Líquida , Exercício Físico , Prostaglandinas/urina , Espectrometria de Massas em Tandem , Adulto , Creatinina/urina , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: It was hypothesized that calcineurin inhibitors suppress vascular cyclooxygenase (COX)-2 and exert a reciprocal influence on in vivo prostacyclin and thromboxane. This could contribute to cardiovascular morbidity in transplanted patients. METHODS: The ability of immunosuppressives to suppress vascular COX-2 expression in vitro was studied in cultured human vascular smooth muscle cells. Blood and urine samples were collected from 28 renal transplant patients before and 2, 4 and 6 h after intake of immunosuppressives and from 11 controls. ELISA was used to measure (1) plasma 6-keto-PGF1alpha and TxB2; (2) urine excretion of PGI-M and TxB(2); (3) 6-keto-PGF1alpha in the whole-blood COX-2 assay; and (4) TxB2 in the whole-blood COX-1 assay. Platelet aggregation was measured optically. RESULTS: COX-2 in cultured vascular smooth muscle cells was suppressed by cyclosporine A (CsA); tacrolimus and rapamycin had no effect. Human renal arteries and vascular smooth muscle expressed calcineurin Abeta and Agamma isoforms. CsA had no effect on plasma 6-keto-PGF1alpha, whole-blood COX-2 activity or PGI-M urine excretion; after rapamycin intake, the former two increased. Plasma TxB2 did not change after drug intake. TxB2 in the COX-1 assay and urine excretion of TxB2 was significantly lower in tacrolimus- and rapamycin-treated patients compared to the CsA group. Platelet aggregation was increased significantly in the CsA group. CONCLUSIONS: Although CsA suppressed COX-2 in cultured vascular smooth muscle cells, systemic prostacyclin was not suppressed by either CsA or tacrolimus in vivo. Rapamycin and tacrolimus may actively suppress platelet and renal thromboxane formation. Differential changes in prostanoids may have implications for long-term cardiovascular hazard in patients treated with immunosuppressives.
Assuntos
Ciclo-Oxigenase 2/metabolismo , Imunossupressores/farmacologia , Transplante de Rim , Rim/efeitos dos fármacos , Rim/enzimologia , 6-Cetoprostaglandina F1 alfa/sangue , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Idoso , Calcineurina/metabolismo , Células Cultivadas , Ciclosporina/farmacologia , Feminino , Humanos , Rim/irrigação sanguínea , Masculino , Pessoa de Meia-Idade , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/enzimologia , Agregação Plaquetária/efeitos dos fármacos , Sirolimo/farmacologia , Tacrolimo/farmacologia , Tromboxano B2/sangue , Tromboxano B2/urina , Adulto JovemRESUMO
BACKGROUND: The precise pathway by which alcohol causes the characteristic features of fetal alcohol spectrum disorders is unknown. Proposed mechanisms for fetal injury from maternal alcohol use include cellular damage from oxidative stress and impaired fetal oxygenation related to maternal systemic vasoconstriction. Our objective was to compare the levels of urinary markers of oxidative stress and systemic vasoconstriction between women consuming large amounts of alcohol during pregnancy and women who did not drink alcohol during pregnancy. METHODS: Pregnant women consuming > or =48 g alcohol per day (n = 29) on average and pregnant women who abstained from alcohol use (n = 39) were identified using detailed interviews and home visits. Random maternal urine specimens were collected. Urinary levels of the oxidative stress marker, 8-isoprostane F2alpha, and of the vasoactive prostaglandin metabolites, 2,3-dinor-6-keto-prostaglandin F1alpha (a vasodilator) and 11-dehydro-thromboxane B2 (a vasoconstrictor), were measured using mass spectrometric methods. All analyte levels were corrected for urinary creatinine. RESULTS: In crude analyses, there was no significant difference in 8-isoprostane F2alpha between pregnant drinkers and nondrinkers (2.16 vs. 2.08 ng/mg creatinine, respectively, p = 0.87). There were no significant differences between the drinking and nondrinking groups in levels of 2,3-dinor-6-keto-prostaglandin F1alpha (1.03 vs. 1.17 ng/mg creatinine, respectively, p = 0.50), 11-dehydro-thromboxane B2 (0.72 vs. 0.59 ng/mg creatinine, respectively, p = 0.21), or the ratio of vasodilatory metabolite to vasoconstrictive metabolite (1.73 vs. 2.72, respectively, p = 0.14). Adjusting for maternal age, marital status, smoking, and gestational age at sampling did not substantially alter the results. CONCLUSION: Our results show no difference in levels of urinary eicosanoid markers of oxidative stress and systemic vasoconstriction between pregnant women who drink heavily and pregnant women who abstain. These findings speak against a role for maternal oxidative stress or systemic vasoconstriction in the pathogenesis of alcohol damage to the fetus.
Assuntos
Consumo de Bebidas Alcoólicas/fisiopatologia , Eicosanoides/urina , Transtornos do Espectro Alcoólico Fetal/etiologia , Isoprostanos/urina , Estresse Oxidativo/fisiologia , Vasoconstrição/fisiologia , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Adolescente , Adulto , Consumo de Bebidas Alcoólicas/efeitos adversos , Estudos de Casos e Controles , Depressores do Sistema Nervoso Central/efeitos adversos , Estudos de Coortes , Dinoprosta/análogos & derivados , Dinoprosta/urina , Etanol/efeitos adversos , Feminino , Transtornos do Espectro Alcoólico Fetal/fisiopatologia , Seguimentos , Humanos , Gravidez , Tromboxano B2/análogos & derivados , Tromboxano B2/urina , Adulto JovemRESUMO
BACKGROUND: In dogs, the effects of immunosuppressive medications on hemostasis are not well known. HYPOTHESIS/OBJECTIVES: The objective was to determine the effects of immunosuppressive medications on primary and secondary hemostasis. Our hypothesis was that cyclosporine and prednisone would increase markers of hypercoagulability and thromboxane synthesis, while azathioprine, mycophenolate mofetil, and leflunomide would have minimal effects on hemostasis. ANIMALS: Eight healthy dogs. METHODS: A randomized, cross-over study used aggregometry, the PFA-100 platelet function analyzer, viscoelastometry, platelet count, and prothrombin and activated partial thromboplastin times to evaluate hemostasis during the administration of prednisone, azathioprine, cyclosporine, mycophenolate mofetil, and leflunomide for 1 week each at standard oral doses. Urine 11-dehydro-thromboxane-B2 (11-dTXB2 ) and 6-keto-prostaglandin-F1α (6-keto-PGF1α ) concentrations, normalized to urine creatinine concentration, were measured. RESULTS: The aggregometry amplitude decreased from 51 ± 21 to 27 ± 14 (P = .002) during leflunomide treatment (ADP activation), but there were no differences in amplitude (P = .240) for any medications when platelets were activated with collagen. For all medications, there were no significant differences in viscoelastometry indices (ACT, P = .666; ClotRate, P = .340; and platelet function, P = .411) and platelet count (P = .552). Compared with pretreatment values, urinary 11-dTXB2 -to-creatinine ratio increased (P = .001) after drug administration (from 3.7 ± 0.6 to 5.6 ± 1.1). Cyclosporine was associated with an increase (P < .001) in the 6-keto-PGF1α -to-creatinine ratio (from 10.3 ± 4.6 to 22.1 ± 5.3). CONCLUSIONS AND CLINICAL IMPORTANCE: Most immunosuppressive drugs do not enhance platelet function or coagulation in healthy dogs, suggesting that these medications might not predispose hypercoagulable dogs to thromboembolism. The results of our study need to be correlated with the clinical outcomes of hypercoagulable dogs.
Assuntos
Hemostasia/efeitos dos fármacos , Imunossupressores/farmacologia , 6-Cetoprostaglandina F1 alfa/urina , Animais , Azatioprina/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Plaquetas/efeitos dos fármacos , Creatinina/urina , Estudos Cross-Over , Ciclosporina/farmacologia , Cães , Feminino , Isoxazóis/farmacologia , Leflunomida , Masculino , Ácido Micofenólico/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Contagem de Plaquetas/veterinária , Prednisona/farmacologia , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
Prostaglandin endoperoxides (PGG2/PGH2), precursors of thromboxane (TX) A2 and prostaglandins, may accumulate sufficiently in the presence of a TXA2 synthase inhibitor to exert biological activity. To address whether this modulates the response to TXA2 synthase inhibition in the setting of thrombosis in vivo, we examined the interaction of a TXA2 synthase inhibitor (U63,557a) and a TXA2/prostaglandin endoperoxide receptor antagonist (L636,499) in a canine model of coronary thrombosis after electrically induced endothelial injury. U63,557a exerted little inhibitory effect in this model despite a marked reduction in serum TXB2 and urinary 2,3-dinor-TXB2, an index of TXA2 biosynthesis. Combination of the two drugs was more effective than either drug alone. The enhanced effect achieved upon addition of the TXA2/prostaglandin endoperoxide receptor antagonist to the TXA2 synthase inhibitor suggests that the response to the latter compound was limited by the proaggregatory effects of prostaglandin endoperoxides. The increased effect of the combination over the receptor antagonist alone may reflect metabolism of PGG2/PGH2 to platelet inhibitory prostaglandins. This is supported by the following findings: (a) urinary 2,3-dinor-6-keto-PGF1 alpha, an index of prostacyclin biosynthesis, increased after administration of the synthase inhibitor, an effect that was exaggerated in the presence of thrombosis; (b) inhibition of arachidonate-induced platelet aggregation by U63,557a was dependent on the formation of a platelet-inhibitory prostaglandin; and (c) pretreatment with aspirin abolished the synergism between these compounds. These studies demonstrate that prostaglandin endoperoxides modulate the response to TXA2 synthase inhibition in vivo and identify a drug combination of potential therapeutic efficacy in the prevention of thrombosis.
Assuntos
Doença das Coronárias/enzimologia , Trombose Coronária/enzimologia , Endoperóxidos de Prostaglandina/farmacologia , Tromboxano-A Sintase/antagonistas & inibidores , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Animais , Dibenzotiepinas/farmacologia , Cães , Masculino , Agregação Plaquetária/efeitos dos fármacos , Endoperóxidos Sintéticos de Prostaglandinas/farmacologia , Tromboxano B2/análogos & derivados , Tromboxano B2/sangue , Tromboxano B2/urinaRESUMO
The clinical use of doxorubicin, an anthracycline chemotherapeutic agent, is limited by cardiotoxicity, particularly when combined with herceptin, an antibody that blocks the HER2 receptor. Doxorubicin induces cyclooxygenase-2 (COX-2) activity in rat neonatal cardiomyocytes. This expression of COX-2 limits doxorubicin-induced cardiac cell injury, raising the possibility that the administration of a prostaglandin may protect the heart during the in vivo administration of doxorubicin. Doxorubicin (15 mg/kg) administered to adult male Sprague Dawley rats induced COX-2 expression and activity in cardiac tissue. Prostacyclin generation measured as the excretion of 2,3-dinor-6-keto-PGF(1alpha) also increased, and this was blocked by a COX-2 inhibitor, SC236. In contrast, administration of a COX-1 inhibitor SC560 at a dose that reduced serum thromboxane B2 by more than 80% did not prevent the doxorubicin-induced increase in prostacyclin generation. Doxorubicin increased cardiac injury, detected as a rise in plasma cardiac troponin T, serum lactate dehydrogenase, and cardiomyocyte apoptosis; this was aggravated by coadministration of SC236 but not SC560. The degree of injury in animals treated with a combination of doxorubicin and SC236 was attenuated by prior administration of the prostacyclin analogue iloprost. These data raise the possibility of protecting the heart during the administration of doxorubicin by prior administration of prostacyclin.
Assuntos
6-Cetoprostaglandina F1 alfa/análogos & derivados , Aspirina/farmacologia , Cardiomiopatias/induzido quimicamente , Inibidores de Ciclo-Oxigenase/toxicidade , Doxorrubicina/toxicidade , Coração/efeitos dos fármacos , Isoenzimas/antagonistas & inibidores , Miocárdio/patologia , Nitrobenzenos/farmacologia , Pirazóis/toxicidade , Sulfonamidas/farmacologia , Sulfonamidas/toxicidade , 6-Cetoprostaglandina F1 alfa/urina , Animais , Apoptose/efeitos dos fármacos , Ácido Araquidônico/farmacologia , Biomarcadores , Cardiomiopatias/metabolismo , Cardiomiopatias/prevenção & controle , Ciclo-Oxigenase 1 , Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase/farmacologia , Doxorrubicina/farmacologia , Indução Enzimática/efeitos dos fármacos , Epoprostenol/biossíntese , Epoprostenol/fisiologia , Iloprosta/uso terapêutico , Isoenzimas/biossíntese , Isoenzimas/genética , Isoenzimas/fisiologia , L-Lactato Desidrogenase/sangue , Masculino , Proteínas de Membrana , Miocárdio/metabolismo , Prostaglandina-Endoperóxido Sintases/biossíntese , Prostaglandina-Endoperóxido Sintases/genética , Prostaglandina-Endoperóxido Sintases/fisiologia , Pirazóis/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/metabolismo , Tromboxano B2/sangue , Fatores de Transcrição/metabolismo , Troponina T/sangueRESUMO
This multicenter, double-blind, randomized, placebo-controlled, parallel-group study assessed renal function during dosing with etoricoxib 90 mg daily, celecoxib 200 mg twice daily, and naproxen 500 mg twice daily. Male and female subjects 60 to 81 years old (n = 85), in sodium balance on a controlled, normal sodium diet, were treated for 15 days. There were no clinically meaningful between-treatment differences in urinary sodium excretion, creatinine clearance, body weight, or serum electrolytes during the 2 weeks of treatment. Etoricoxib and celecoxib had no effect on the urinary thromboxane metabolite, 11-dehydrothromboxane B(2), while significantly decreasing the urinary prostacyclin metabolite, 2,3-dinor-6-keto PGF(1alpha). Decreases were greater for both metabolites following naproxen. Ambulatory systolic blood pressures were significantly higher than placebo for all treatments, with moderately greater increases for etoricoxib relative to other active treatments on day 14. Ambulatory diastolic blood pressures were significantly higher than placebo for etoricoxib and naproxen but not for celecoxib.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Piridinas/farmacologia , Sódio/urina , Sulfonas/farmacologia , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Administração Oral , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/efeitos adversos , Peso Corporal/efeitos dos fármacos , Celecoxib , Constipação Intestinal/induzido quimicamente , Creatinina/sangue , Creatinina/urina , Diarreia/induzido quimicamente , Relação Dose-Resposta a Droga , Método Duplo-Cego , Eletrólitos/sangue , Etoricoxib , Feminino , Cefaleia/induzido quimicamente , Humanos , Masculino , Pessoa de Meia-Idade , Naproxeno/administração & dosagem , Naproxeno/efeitos adversos , Naproxeno/farmacologia , Potássio/urina , Prostaglandinas/urina , Pirazóis/administração & dosagem , Pirazóis/efeitos adversos , Pirazóis/farmacologia , Piridinas/administração & dosagem , Piridinas/efeitos adversos , Sulfonamidas/administração & dosagem , Sulfonamidas/efeitos adversos , Sulfonamidas/farmacologia , Sulfonas/administração & dosagem , Sulfonas/efeitos adversos , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
Our study aimed to determine the role of cyclooxygenase-2 in the release of prostaglandin-(PG)-I2 following mesenteric traction during abdominal surgery. In a prospective double-blind, randomized, placebo-controlled study, 40 patients electively scheduled for non-laparoscopic abdominal surgery, were pretreated with the cyclooxygenase-2 inhibitor parecoxib (n=20) or placebo (n=20). Heart rate, arterial blood pressure, oxygenation ratio and plasma concentrations of the stable PGI2-metabolite 6-keto-PGF1alpha were compared between groups before injection of parecoxib (-40 min), immediately before mesenteric traction (0 min), and 5, 10, and 30 min thereafter. In addition, plasma concentrations of valdecoxib, the active metabolite of the prodrug parecoxib, were determined. Plasma concentrations of 6-keto-PGF1alpha and heart rate increased in both groups after mesenteric traction. There were no significant differences between groups at individual times in heart rate, arterial blood pressure and plasma concentrations of 6-keto-PGF1alpha. Oxygenation ratio decreased after 10 and 30 min following mesenteric traction in the parecoxib group with a significant difference between treatment groups at 10 and 30 min. Plasma concentrations of valdecoxib revealed therapeutic values. Our data indicate that PGI2 release following mesenteric traction is mediated by cyclooxygenase-1.
Assuntos
Abdome/cirurgia , Ciclo-Oxigenase 1/metabolismo , Epoprostenol/metabolismo , Complicações Intraoperatórias/prevenção & controle , Isoxazóis/uso terapêutico , 6-Cetoprostaglandina F1 alfa/sangue , 6-Cetoprostaglandina F1 alfa/urina , Pressão Sanguínea/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/administração & dosagem , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Método Duplo-Cego , Epoprostenol/sangue , Epoprostenol/urina , Feminino , Rubor/etiologia , Rubor/fisiopatologia , Rubor/prevenção & controle , Frequência Cardíaca/efeitos dos fármacos , Humanos , Hipotensão/etiologia , Hipotensão/fisiopatologia , Hipotensão/prevenção & controle , Complicações Intraoperatórias/etiologia , Complicações Intraoperatórias/fisiopatologia , Isoxazóis/administração & dosagem , Isoxazóis/sangue , Laparoscopia , Masculino , Pessoa de Meia-Idade , Oxigênio/metabolismo , Estudos Prospectivos , Sulfonamidas/sangue , Procedimentos Cirúrgicos Operatórios/efeitos adversos , Procedimentos Cirúrgicos Operatórios/métodos , Taquicardia/etiologia , Taquicardia/fisiopatologia , Taquicardia/prevenção & controle , Fatores de Tempo , Resultado do TratamentoRESUMO
Gestational choriocarcinoma metastasizes rapidly, in which process the vasoactive prostanoids may be significant. We therefore compared the urinary excretion of prostacyclin and thromboxane A2 (TxA2) metabolites in 19 women with gestational choriocarcinoma and 20 healthy age-matched women by assessing spot urine samples for 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and 2,3-dinor-6-keto-prostaglandin F1 alpha (2,3-dinor-6-keto-PGF1 alpha) (degradation products of prostacyclin) as well as for thromboxane B2 (TxB2) and 2,3-dinor-TxB2 (degradation products of TxA2) by high-pressure liquid chromatography, followed by radioimmunoassay; the data were related to urinary creatinine concentration. The urinary output of 6-keto-PGF1 alpha [29.56 +/- 7.0 versus 25.08 +/- 3.91 ng/mmol creatinine (SE)] in patients with choriocarcinoma was normal, but that of 2,3-dinor-6-keto-PGF1 alpha in cancer patients was higher than in controls (24.44 +/- 5.20 versus 14.84 +/- 1.94, P less than 0.02), as was that of TxB2 (22.72 +/- 4.69 versus 9.69 +/- 1.52, P less than 0.001) and 2,3-dinor-TxB2 (114.21 +/- 30.81 versus 51.81 +/- 10.40, P less than 0.01). The ratio of net prostacyclin output (6-keto-PGF1 alpha plus 2,3-dinor-6-keto-PGF1 alpha) to the net TxA2 output (TxB2 plus 2,3-dinor-TxB2) in cancer patients [0.52 +/- 0.1 (SE)] was lower (P less than 0.03) than in the controls (0.83 +/- 0.1), and in an inverse relation (r = -0.54, P less than 0.05) to the scoring index of poor prognosis for the disease. We conclude that the prostanoid excess in gestational trophoblastic disease, as evidenced for the first time in this study, may originate from choriocarcinoma cells, or may be a paraneoplastic phenomenon, and we conclude also that TxA2 excess may contribute to the tumor growth and/or formation of metastases.
Assuntos
6-Cetoprostaglandina F1 alfa/urina , Biomarcadores Tumorais/urina , Coriocarcinoma/urina , Epoprostenol/metabolismo , Tromboxano A2/urina , Neoplasias Uterinas/urina , 6-Cetoprostaglandina F1 alfa/análogos & derivados , Adulto , Feminino , Humanos , Gravidez , Valores de Referência , Tromboxano A2/metabolismo , Tromboxano B2/análogos & derivados , Tromboxano B2/urinaRESUMO
PURPOSE: Obstructive sleep apnea increases the risk of cardiovascular diseases. Alternations in prostacyclin and thromboxane concentrations and balance could constitute one of mechanisms linking sleep apnea and cardiovascular events. Thus we aimed to assess the concentrations of 6-keto-prostaglandin F1α (6-keto-PGF1α) (metabolite of prostacyclin) and thromboxane B2 (TXB2) (metabolite of thromboxane A2) in urine and blood of obstructive sleep apnea patients and controls (snoring subjects with otherwise normal polysomnogram). MATERIAL AND METHODS: Overnight urine and morning blood samples were taken from subjects and controls at baseline and in sleep apnea group during continuous positive airway pressure (CPAP) treatment. Samples were analyzed using mass chromatography/gas spectrometry. RESULTS: We analyzed data from 26 obstructive sleep apnea subjects (mean apnea-hypopnea index 45.4±17.3) and 22 well-matched controls. At baseline sleep apnea patients, when compared to controls, have higher 6-keto-PGF1α in urine (0.89±0.15 vs 0.34±0.06, p=0.01) and blood (24.49±1.54 vs 19.70±1.77, p=0.04). TXB2 levels in urine and blood were not different across groups. CPAP treatment significantly decreased 6-keto-PGF1α in urine (0.92±0.17 vs 0.22±0.10, p=0.04), but not in blood. TXB2 levels during CPAP treatment did not change significantly. CONCLUSIONS: These results suggest augmented systemic prostacyclin production in obstructive sleep apnea patients, which potentially could constitute a protective mechanism against detrimental effects of sleep apnea.
Assuntos
Epoprostenol/biossíntese , Apneia Obstrutiva do Sono/metabolismo , Tromboxanos/biossíntese , 6-Cetoprostaglandina F1 alfa/sangue , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Idoso , Estudos de Casos e Controles , Pressão Positiva Contínua nas Vias Aéreas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Apneia Obstrutiva do Sono/sangue , Apneia Obstrutiva do Sono/urina , Tromboxano B2/sangue , Tromboxano B2/urinaRESUMO
Excretion of 2,3-dinor-6-ketoprostaglandin F1 alpha and 2,3-dinorthromboxane B2, the main urinary metabolites of prostacyclin and thromboxane, was evaluated by gas chromatography-mass spectroscopy and radioimmunoassay, respectively, at various conditions in man. In healthy young males excretion of 2,3-dinor-6-ketoprostaglandin F1 alpha was of little variability, whereas urinary 2,3-dinorthromboxane B2 showed marked interindividual but moderate intraindividual variations. The ratio of urinary 2,3-dinorthromboxane B2 to thromboxane B2 in young males was about 15. Excretion of 2,3-dinor-6-ketoprostaglandin F1 alpha in women of reproductive age was higher (155 +/- 23 ng/g creatinine, P less than 0.005) than in postmenopausal women (97 +/- 24 ng/g creatinine) and in men (78 +/- 7.6 ng/g creatinine) and increased significantly during pregnancy (1st trimester 230 +/- 50 ng/g creatinine; 3rd trimester 522 +/- 53 ng/g creatinine). Urinary 2,3-dinorthromboxane B2 showed no gender differences and no directed change was observed during pregnancy. In neonates urinary 2,3-dinorthromboxane B2 (6.328 +/- 1.210 ng/g creatinine) was high in their 3rd day of life and decreased rapidly thereafter. This pattern paralleled the behavior of 6-ketoprostaglandin F1 alpha. In young male smokers and non-smokers excretion of 2,3-dinor-6-ketoprostaglandin F1 alpha was not significantly different, whereas urinary 2,3-dinorthromboxane B2 was elevated in smokers (609 +/- 61 versus 351 +/- 41 ng/g creatinine, P less than 0.001). Values are mean +/- S.E.
Assuntos
6-Cetoprostaglandina F1 alfa/análogos & derivados , Tromboxano B2/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Adulto , Envelhecimento , Pré-Escolar , Creatinina/urina , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Recém-Nascido , Cinética , Tromboxano B2/urinaRESUMO
In vivo biosynthesis of thromboxane and prostacyclin is currently evaluated by measuring urinary excretion of selected metabolites. Urinary thromboxane B2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) (non-enzymatic hydrolysis products of thromboxane and prostacyclin) are thought to derive from renal biosynthesis of the parent compounds, while enzymatic metabolites such as 2,3-dinor-TXB2 and 2,3-dinor-6-keto-PGF1 alpha appear to be mainly derived from systemic (platelet) thromboxane and (vascular) prostacyclin, respectively. Using immunoaffinity extraction and high-resolution gas chromatography-negative ion chemical ionization mass spectrometry (HRGC-NICIMS), we measured the paired excretion of non-enzymatic and enzymatic metabolites of thromboxane and prostacyclin in healthy subjects before, during and after an eight-day schedule of oral low-dose aspirin (30 mg/day), a treatment known to inhibit platelet and perhaps vascular but not renal cyclooxygenase. Low-dose aspirin cumulatively reduced urinary excretion of TXB2 and 2,3-dinor-TXB2 (about 80% inhibition on day 8 of aspirin treatment, P less than 0.01), as well as 6-keto-PGF1 alpha and 2,3-dinor-6-keto-PGF1 alpha (about 45% inhibition on day 8 of aspirin treatment, P less than 0.01). Excretion of all metabolites recovered slowly after aspirin withdrawal. Urinary PGE2, taken as an index of renal cyclooxygenase activity, was not inhibited by aspirin. A highly significant correlation was found between paired excretion values of non-enzymatic vs. enzymatic metabolites of thromboxane and prostacyclin in all individuals studied (TXB2 vs. 2,3-dinor-TXB2 (r = 0.91 +/- 0.03); 6-keto-PGF1 alpha vs. 2,3-dinor-6-keto-PGF1 alpha (r = 0.92 +/- 0.06], irrespective of aspirin treatment. TXB2/2,3-dinor-TXB2 and 6-keto-PGF1 alpha/2,3-dinor-6-keto-PGF1 alpha mean ratios remained unchanged throughout the experiment. These data do not support the view that urinary TXB2 and 6-keto-PGF1 alpha derive mainly from renal biosynthesis in healthy subjects, but rather suggest that they may represent a fraction of systemic (platelet) thromboxane and (vascular) prostacyclin escaping metabolism. These data also suggest that chronic low-dose aspirin may partly inhibit vascular prostacyclin in addition to platelet thromboxane biosynthesis.