RESUMEN
Grape consumption acts on the immune system to produce antioxidant and anti-inflammatory effects. Since immune activity demonstrates circadian rhythmicity, with peak activity occurring during waking hours, the timing of grape intake may influence the magnitude of its antioxidant effect. This study followed a 2 × 2 factorial randomized, controlled design wherein healthy men and women (n = 32) consumed either a grape or placebo drink with a high-fat meal in the morning or evening. Urine was collected for measurements of biomarkers of oxidative stress and grape metabolites at baseline and post-meal at hour 1 and hours 1-6. F-2 isoprostane levels showed main effects of time period (baseline < hour 1 < hours 1-6, p < 0.0001), time (a.m. > p.m., p = 0.008) and treatment (placebo > grape, p = 0.05). Total F2-isoprostane excretion expressed as % baseline was higher in the a.m. vs. p.m. (p = 0.004) and in the a.m. placebo vs. all other groups (p < 0.05). Tartaric acid and resveratrol excretion levels were higher in the grape vs. placebo group (p < 0.05) but were not correlated with F-2 isoprostane levels. The findings support a protective effect of grape consumption against morning sensitivity to oxidative stress.
Asunto(s)
Antioxidantes , Vitis , Masculino , Humanos , Femenino , Antioxidantes/farmacología , Estrés Oxidativo , F2-Isoprostanos/farmacología , Isoprostanos/farmacologíaRESUMEN
Renal ischemia-reperfusion injury (IRI) is a critical health concern that aggravates the pathophysiology of acute kidney injury (AKI), leading to high mortality rates in intensive care units. Cardamonin is a natural compound with anti-inflammatory and antioxidant properties. The current study aimed to evaluate the renoprotective impact of cardamonin against AKI induced by renal IRI. Male rats (n=5 per group) were divided into four groups: the sham group underwent anesthesia and abdominal incision only; the control group experienced bilateral renal artery clamping for 30 minutes followed by 2 hours of reperfusion; the vehicle group received the cardamonin vehicle 30 minutes before ischemia induction; and the cardamonin group was administered 5 mg/kg of cardamonin 30 minutes before ischemia. Blood urea nitrogen (BUN) and creatinine were measured to assess the renal function. Tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1ß), interleukin-6 (IL-6), caspase 3, and F2-isoprostane were assessed in renal tissues. Kidney injury was examined using the hematoxylin and eosin stain method. Compared to the sham group, the control group exhibited significantly higher levels of BUN, creatinine, TNF-α, IL-1ß, IL-6, F2-isoprostane, and caspase 3 in renal tissues, along with severe kidney injury as evidenced by histological analysis. Compared to the control group, pretreatment with cardamonin resulted in a significant reduction in these biomarkers and alleviated renal damage. Cardamonin had renoprotective effects against renal ischemia and reperfusion injury via modulating inflammation, oxidative stress, and apoptosis pathways.
Asunto(s)
Lesión Renal Aguda , Chalconas , Daño por Reperfusión , Ratas , Masculino , Animales , Caspasa 3/metabolismo , Caspasa 3/farmacología , Factor de Necrosis Tumoral alfa , Interleucina-6 , Creatinina , F2-Isoprostanos/metabolismo , F2-Isoprostanos/farmacología , Ratas Sprague-Dawley , Riñón , Isquemia/tratamiento farmacológico , Lesión Renal Aguda/tratamiento farmacológico , Lesión Renal Aguda/etiología , Lesión Renal Aguda/prevención & control , Apoptosis , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/prevención & control , Estrés Oxidativo , Reperfusión , Inflamación/tratamiento farmacológico , Inflamación/patologíaRESUMEN
PURPOSE: Carotenoids may protect against chronic diseases including cancer and cardiometabolic disease by mitigating oxidative stress and/or inflammation. We cross-sectionally evaluated associations between carotenoids and biomarkers of oxidative stress or inflammation. METHODS: From 2003 to 2009, the Sister Study enrolled 50,884 breast cancer-free US women aged 35-74. Post-menopausal participants (n = 512) were randomly sampled to measure carotenoids and biomarkers of oxidative stress. Dietary carotenoid consumption was assessed using a validated 110-item Block 1998 food frequency questionnaire; use of ß-carotene-containing supplements was also assessed. Plasma carotenoids were quantified, adjusting for batch. Urinary markers of lipid peroxidation, 8-iso-prostaglandin F2α (8-iso-PGF2α) and its metabolite (8-iso-PGF2α-M) were also measured. Since the biomarker 8-iso-PGF2α can reflect both oxidative stress and inflammation, we used a modeled 8-iso-PGF2α to prostaglandin F2α ratio approach to distinguish effects reflecting oxidative stress versus inflammation. Multivariable linear regression was used to assess the associations of dietary and plasma carotenoids with the estimated biomarker concentrations. RESULTS: Total plasma carotenoids were inversely associated with 8-iso-PGF2α-M concentrations (P for trend across quartiles = 0.009). Inverse trends associations were also seen for α-carotene and ß-carotene. In contrast, lutein/zeaxanthin showed associations with both 8-iso-PGF2α and 8-iso-PGF2α-M concentrations. The inverse association for total carotenoids appeared to be specific for oxidative stress (chemical 8-iso-PGF2α; Phighest vs. lowest quartile = 0.04 and P for trend across quartiles = 0.02). The pattern was similar for α-carotene. However, lutein/zeaxanthin tended to have a stronger association with enzymatic 8-iso-PGF2α, suggesting an additional anti-inflammatory effect. Supplemental ß-carotene was inversely associated with both 8-iso-PGF2α and 8-iso-PGF2α-M concentrations, as well as with both chemical and enzymatic 8-iso-PGF2α. Dietary carotenoids were not associated with either biomarker. CONCLUSION: Plasma carotenoids and supplemental ß-carotene were associated with lower concentrations of 8-iso-PGF2α metabolite. Plasma carotenoids associations may reflect antioxidant effects.
Asunto(s)
F2-Isoprostanos , Isoprostanos , Biomarcadores , Carotenoides , Dinoprost , F2-Isoprostanos/farmacología , Femenino , Humanos , Inflamación/metabolismo , Luteína , Estrés Oxidativo , Zeaxantinas/metabolismo , Zeaxantinas/farmacología , beta CarotenoRESUMEN
Recent epidemiological studies suggest a strong association between exposure to environmental contaminants, including organochlorine (OC) insecticides or their metabolites, and development of pathologies, such as atherosclerosis, in which oxidative stress plays a significant etiological role. Biomarkers of systemic oxidative stress have the potential to link production of reactive oxygen species (ROS), which are formed as a result of exposure to xenobiotic toxicants, and underlying pathophysiological states. Measurement of F2-isoprostane concentrations in body fluids is the most accurate and sensitive method currently available for assessing in vivo steady-state oxidative stress levels. In the current study, urinary concentrations of F2-isoprostanes and serum levels of persistent OC compounds p,p'-dichlorodiphenyldichloroethene (DDE), trans-nonachlor (a component of the technical chlordane mixture), and oxychlordane (a chlordane metabolite) were quantified in a cross-sectional study sample and the association of these factors with a clinical diagnosis of atherosclerosis determined. Urinary isoprostane levels were not associated with atherosclerosis or serum concentrations of OC compounds in this study sample. However, occurrence of atherosclerosis was found to be associated with serum trans-nonachlor levels. DDE and oxychlordane were not associated with atherosclerosis. This finding supports current evidence that exposure to environmental factors is a risk factor for atherosclerosis, in addition to other known risk factors.
Asunto(s)
Aterosclerosis/epidemiología , Contaminantes Ambientales/sangre , F2-Isoprostanos/sangre , Hidrocarburos Clorados/sangre , Insecticidas/sangre , Anciano , Anciano de 80 o más Años , Aterosclerosis/inducido químicamente , Biomarcadores/sangre , Estudios Transversales , F2-Isoprostanos/farmacología , Femenino , Humanos , Hidrocarburos Clorados/farmacología , Insecticidas/farmacología , Masculino , Persona de Mediana Edad , Mississippi/epidemiología , Estrés Oxidativo/efectos de los fármacos , Factores de RiesgoRESUMEN
Effects of propofol, an intravenous anesthetic agent that exerts potent antioxidant properties, were investigated in an experimental model of cardiac arrest and cardiopulmonary resuscitation. An extended cardiac arrest with 15 randomized piglets was studied to assess the effect of propofol or its solvent intralipid as the control group. Oxidative stress (as measured by a major F(2)-isoprostane) and inflammation (a major metabolite of PGF(2α)) were evaluated in addition to the hemodynamic evaluation, protein S-100ß and in situ tissue brain damage by immunochemistry at sacrifice after 3h of reperfusion following cardiac arrest and restoration of spontaneous circulation (ROSC). ROSC increased jugular bulb plasma levels of F(2)-isoprostane and PGF(2α) metabolite significantly more in controls than in the propofol-treated group. In situ tissue damage after ischemia-reperfusion was variable among the pigs at sacrifice, but tended to be greater in the control than the propofol-treated group. Propofol significantly reduced an ROSC-mediated oxidative stress in the brain.
Asunto(s)
Anestésicos Intravenosos/uso terapéutico , Reanimación Cardiopulmonar/efectos adversos , Depuradores de Radicales Libres/uso terapéutico , Propofol/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Animales , Encéfalo/metabolismo , Circulación Cerebrovascular/efectos de los fármacos , Modelos Animales de Enfermedad , F2-Isoprostanos/farmacología , Venas Yugulares/efectos de los fármacos , Oxidación-Reducción , Estrés Oxidativo , Daño por Reperfusión/etiología , Daño por Reperfusión/metabolismo , Porcinos/metabolismoRESUMEN
15-F2(t)-isoprostane (15-F2(t)-IsoP), an oxidation product of arachidonic acid (AA), affects vascular and platelet function; however, the bioactivity of other fatty acids oxidation products is unknown. This paper studied rat aortic vascular reactivity and human platelet aggregation in response to 14 oxidation products of AA, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and α-linolenic acid (ALA) compared with 15-F2(t)-IsoP. It also compared the F2(t)-IsoPs profile in human platelets. EPA-derived 15-F3(t)-IsoP constricted rat aorta less than 15-F2(t)-IsoP, but none of the other oxidation products affected vascular reactivity. Only 15-F2(t)-IsoP (10â»4 M) directly affected platelet aggregation. 15-F3(t)-IsoP, ent-16-F1-phytoprostane (from ALA) and isofurans A and B (from AA) inhibited reversible aggregation to U46619. Unlike plasma, the platelet profile of F2-IsoP showed that 8-F(2t)-IsoP were higher than 15-F2(t)-IsoP. Unlike 15-F2(t)-IsoP, the test compounds derived from fatty acids oxidation did not affect vascular or platelet function. Elevated platelet 8-F2(t)-IsoP could limit 15-F2(t)-IsoP-induced aggregation under conditions of oxidant stress.
Asunto(s)
Aorta/efectos de los fármacos , Ácido Araquidónico/farmacología , Ácidos Docosahexaenoicos/farmacología , Ácido Eicosapentaenoico/farmacología , Agregación Plaquetaria/efectos de los fármacos , Ácido alfa-Linolénico/farmacología , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Adulto , Anciano , Animales , Aorta/fisiología , Plaquetas/metabolismo , Ciclopentanos/farmacología , F2-Isoprostanos/farmacología , Ácidos Grasos Insaturados/farmacología , Humanos , Masculino , Persona de Mediana Edad , Oxidación-Reducción , Estrés Oxidativo , Ratas , Ratas Wistar , Adulto JovenRESUMEN
Recently, we reported that reactive oxygen species (ROS) generated by NADPH oxidase (NOX) contribute to aberrant responses in pulmonary resistance arteries (PRAs) of piglets exposed to 3 days of hypoxia (Am J Physiol Lung Cell Mol Physiol 295: L881-L888, 2008). An objective of the present study was to determine whether NOX-derived ROS also contribute to altered PRA responses at a more advanced stage of pulmonary hypertension, after 10 days of hypoxia. We further wished to advance knowledge about the specific NOX and antioxidant enzymes that are altered at early and later stages of pulmonary hypertension. Piglets were raised in room air (control) or hypoxia for 3 or 10 days. Using a cannulated artery technique, we found that treatments with agents that inhibit NOX (apocynin) or remove ROS [an SOD mimetic (M40403) + polyethylene glycol-catalase] diminished responses to ACh in PRAs from piglets exposed to 10 days of hypoxia. Western blot analysis showed an increase in expression of NOX1 and the membrane fraction of p67phox. Expression of NOX4, SOD2, and catalase were unchanged, whereas expression of SOD1 was reduced, in arteries from piglets raised in hypoxia for 3 or 10 days. Markers of oxidant stress, F(2)-isoprostanes, measured by gas chromatography-mass spectrometry, were increased in PRAs from piglets raised in hypoxia for 3 days, but not 10 days. We conclude that ROS derived from some, but not all, NOX family members, as well as alterations in the antioxidant enzyme SOD1, contribute to aberrant PRA responses at an early and a more progressive stage of chronic hypoxia-induced pulmonary hypertension in newborn piglets.
Asunto(s)
Hipertensión Pulmonar/metabolismo , Hipoxia , NADPH Oxidasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Animales Recién Nacidos , Antioxidantes/farmacología , Catalasa/metabolismo , F2-Isoprostanos/farmacología , Hipertensión Pulmonar/etiología , Immunoblotting , Luminiscencia , Manganeso , NADPH Oxidasa 1 , Compuestos Organometálicos/farmacología , Fosfoproteínas/metabolismo , Arteria Pulmonar/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1 , Porcinos , Resistencia Vascular/efectos de los fármacosRESUMEN
F(2)-isoprostanes are not just markers of oxidative stress. The discovery of F(2)-isoprostanes (F(2)-IsoPs) as specific and reliable markers of oxidative stress in vivo is briefly summarized here. F(2)-IsoPs are also agonists of important biological effects, such as the vasoconstriction of renal glomerular arterioles, the retinal vessel, and the brain microcirculature. In addition to the F(2)-IsoPs, E(2)- and D(2)-IsoPs can be formed by rearrangement of H(2)-IsoP endoperoxides and can give rise to cyclopentenone IsoPs, which are very reactive alpha,beta-unsaturated aldehydes. The same type of reactivity is also shown by acyclic gamma-ketoaldehydes formed as products of the IsoP pathway. Because previous studies suggested a relation between oxidative stress and collagen hyperproduction, it was investigated whether collagen synthesis is induced by F(2)-IsoPs, the most proximal products of lipid peroxidation. In contrast to aldehydes, F(2)-IsoPs act through receptors able to elicit definite signal transduction pathways. In a rat model of carbon tetrachloride-induced hepatic fibrosis, plasma F(2)-IsoPs were markedly elevated for the entire experimental period; hepatic collagen content was also increased. When hepatic stellate cells from normal liver were cultured up to activation (expression of smooth muscle alpha-actin) and then treated with F(2)-IsoPs in the concentration range found in the in vivo studies (10(-9) to 10(-8) M), a striking increase in DNA synthesis, cell proliferation, and collagen synthesis was observed. Total collagen content was similarly increased. All these stimulatory effects were reversed by the specific antagonist of the thromboxane A(2) receptor, SQ 29 548, whereas the receptor agonist, I-BOP, also had a stimulatory effect. Therefore F(2)-IsoPs generated by lipid peroxidation in hepatocytes may mediate hepatic stellate cell proliferation and collagen hyperproduction seen in hepatic fibrosis.
Asunto(s)
F2-Isoprostanos/química , F2-Isoprostanos/metabolismo , Cirrosis Hepática/etiología , Hígado/metabolismo , Estrés Oxidativo , Animales , Biomarcadores/química , Biomarcadores/metabolismo , Colágeno/metabolismo , F2-Isoprostanos/farmacología , Humanos , Peroxidación de Lípido , Hígado/efectos de los fármacos , Hígado/patología , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Fibrosis Pulmonar/etiología , Ratas , Receptores de Prostaglandina/metabolismoRESUMEN
After a brief introduction to oxidative stress, the discovery of F(2)-isoprostanes as specific and reliable markers of oxidative stress is described. Isoprostanes are also agonists of important biological effects. Since a relation between oxidative stress and collagen hyperproduction has been previously suggested and since lipid peroxidation products have been proposed as possible mediators of liver fibrosis, we investigated whether collagen synthesis is induced by F(2)-isoprostanes the most proximal products of lipid peroxidation. In a rat model of carbon tetrachloride-induced hepatic fibrosis, plasma isoprostanes were markedly elevated for the entire experimental period; hepatic collagen content was also increased. When hepatic stellate cells from normal liver were cultured up to activation (expression of alpha-smooth muscle-alpha actin) and then treated with F(2)-isoprostanes in the concentration range found in the in vivo studies (10(-9)-10(-8)M), a striking increase in DNA synthesis, in cell proliferation and in collagen synthesis was observed. Moreover, F(2)-isoprostanes increased the production of transforming growth factor-beta1 by U937 cells, assumed as a model of Kupffer cells or liver macrophages. The data suggest the possibility that F(2)-isoprostanes generated by lipid peroxidation in hepatocytes mediate hepatic stellate cell proliferation and collagen hyperproduction seen in hepatic fibrosis.
Asunto(s)
Isoprostanos/sangre , Cirrosis Hepática/sangre , Animales , Proliferación Celular/efectos de los fármacos , Colágeno/metabolismo , F2-Isoprostanos/metabolismo , F2-Isoprostanos/farmacología , Humanos , Isoprostanos/metabolismo , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Estrés Oxidativo/efectos de los fármacosRESUMEN
Carbon tetrachloride (CCl4)-induced hepatic fibrosis has been considered to be linked to oxidative stress and mediated by aldehydic lipid peroxidation products. In the present study, we investigated whether collagen synthesis is induced by F2-isoprostanes, the most proximal products of lipid peroxidation and known mediators of important biological effects. By contrast with aldehydes, F2-isoprostanes act through receptors able to elicit definite signal transduction pathways. In a rat model of CCl4-induced hepatic fibrosis, plasma F2-isoprostanes were markedly elevated for the entire experimental period; hepatic collagen content also increased. When hepatic stellate cells (HSCs) from normal liver were cultured with F2-isoprostanes in the concentration range found in the in vivo studies (10(-9)-10(-8) M), a striking increase in DNA synthesis (reversed by the thromboxane A2 antagonist SQ 29 548), in cell proliferation and in collagen synthesis was observed. Total collagen content was similarly increased. Moreover, F2-isoprostanes markedly increased the production of transforming growth factor-beta1 by U937 cells, considered a model of liver macrophages. The data provide evidence for the possibility that F2-isoprostanes generated by lipid peroxidation in hepatocytes mediate HSC proliferation and collagen production seen in hepatic fibrosis.
Asunto(s)
Colágeno/biosíntesis , F2-Isoprostanos/farmacología , Hepatocitos/efectos de los fármacos , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/patología , Animales , Tetracloruro de Carbono/administración & dosificación , Tetracloruro de Carbono/toxicidad , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Colágeno/análisis , ADN/biosíntesis , Relación Dosis-Respuesta a Droga , F2-Isoprostanos/sangre , Fibrosis/patología , Humanos , Inyecciones Intraperitoneales , Hígado/citología , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta1 , Células U937RESUMEN
F2-isoprostanes (F2-IsoP's) are biologically active prostanoids formed by free radical-mediated peroxidation of arachidonic acid. Four different F2-IsoP regioisomers (5-, 8-, 12-, and 15-series), each comprising eight racemic diastereomers, total 64 compounds. Information regarding the biological activity of IsoP's is largely limited to 15-F2t-IsoP (8-iso-PGF2alpha). We recently demonstrated that 15-F2t-IsoP and its metabolite, 2,3-dinor-5,6-dihydro-15-F2t-IsoP, evoked vasoconstriction and TXA2 generation in retina and brain microvasculature. We have now examined and compared the biological activities of a series of recently synthesized new 5-, 12-, and 15-series F2-IsoP isomers in pig retinal and brain microvasculature. We hereby show that other 15-series F2-IsoP isomers, 15-epi-15-F2t-IsoP, ent-15-F2t-IsoP, and ent-15-epi-15-F2t-IsoP, are also potent vasoconstrictors. The 12-series isomers tested, 12-F2t-IsoP and 12-epi-12-F2t-IsoP, also caused marked vasoconstriction. Of the 5-series isomers tested, 5-F2t-IsoP and 5-epi-5-F2t-IsoP possessed no vasomotor properties, whereas ent-5-F2t-IsoP caused modest vasoconstriction. The vasoconstriction of ent-5-F2t-IsoP, 12-F2t-IsoP, and 12-epi-12-F2t-IsoP was abolished by removal of the endothelium, by TXA2 synthase and receptor inhibitor (CGS12970, L670,596), and by receptor-mediated Ca2+ channel blockade (SK & F96365); correspondingly, these isomers increased TXB2 formation by activating Ca2+ influx (detected with fura 2-AM) through non-voltage-dependent receptor-mediated Ca2+ entry (SK & F96365 sensitive) in endothelial cells. In conclusion, as seen with 15-F2t-IsoP, ent-5-F2t-IsoP, 12-F2t-IsoP, and 12-epi-12-F2t-IsoP constricted both retinal and brain microvessels by inducing endothelium-dependent TXA2 synthesis. These new findings broaden the scope of our understanding regarding the potential involvement of F2-IsoP's as mediators of oxidant injury.
Asunto(s)
Encéfalo/irrigación sanguínea , F2-Isoprostanos/química , F2-Isoprostanos/farmacología , Microcirculación/efectos de los fármacos , Retina/fisiología , Vasoconstricción/efectos de los fármacos , Animales , Calcio/metabolismo , Células Cultivadas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiología , F2-Isoprostanos/síntesis química , Técnicas In Vitro , Isomerismo , Microcirculación/fisiología , Porcinos , Tromboxano A2/metabolismoRESUMEN
BACKGROUND AND OBJECTIVES: 8-isoprostane is a marker of oxidative stress in vivo and increased plasma and urine levels are found in patients with vascular disease and in atherosclerotic plaques. Inflammatory chemokines such as interleukin (IL)-8 seem to play an important pathogenic role in atherogenesis. We therefore investigated the effects of 8-isoprostane on the expression of inflammatory chemokines with consciousness on IL-8 (mRNA and protein) in human macrophages. In addition, we studied the involvement of mitogen-activated protein kinases (ERK 1/2 and p38 MAPK) and nuclear factor-kappaB (NF-kappaB) in this process. METHODS AND RESULTS: 8-isoprostane (10 microM) induced IL-8 expression (mRNA and protein), measured by real-time quantitative RT-PCR and enzyme immunoassay, respectively, in both THP-1 macrophages and human monocyte-derived macrophages. Moreover, 8-isoprostane increased mRNA expression of macrophage inflammatory protein-1alpha as determined by RNase protection assay. In this process, 8-isoprostane induced the activation of two major MAP-kinases; ERK 1/2 and p38 MAPK. Furthermore, the ERK 1/2 inhibitor, PD98059, and the p38 MAPK inhibitor, SB203580, markedly reduced 8-isoprostane-induced IL-8 expression (mRNA and protein), while inhibition of NF-kappaB activation and translocation had no significant effect on IL-8 expression. CONCLUSIONS: We show that 8-isoprostane increases IL-8 expression in human macrophages involving both ERK 1/2 and p38 MAPK, but not NF-kappaB signaling pathway. These findings further support a link between oxidative stress/lipid peroxidation and inflammation in human macrophages and suggest a role for 8-isoprostane in this process. This 8-isoprostane-induced chemokine expression might be involved in the pathogenesis of atherosclerosis as well as other inflammatory disorders.
Asunto(s)
Dinoprost/análogos & derivados , F2-Isoprostanos/farmacología , Interleucina-8/metabolismo , Macrófagos/enzimología , Línea Celular , Células Cultivadas , Activación Enzimática , Expresión Génica/efectos de los fármacos , Humanos , Inflamación , Interleucina-8/análisis , Interleucina-8/genética , Sistema de Señalización de MAP Quinasas , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , ARN Mensajero/análisis , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
We investigated the effects of five different isoprostanes (8-iso PGE1, 8-iso PGE2, 8-iso PGF1alpha, 8-iso PGF2alpha and 8-iso PGF2beta) on vasomotor tone in human and porcine bronchial arterial tissues. In the human bronchial arteries, 8-iso PGE2 and 8-iso PGF2alpha evoked powerful constrictions (magnitudes several fold greater than the responses to high millimolar KCl) with negative log concentration causing 50% excitation (EC50) values of 6.8 and 6.5, respectively; 8-iso PGE1 was less potent (EC50 not calculated, since a clear peak contraction was not obtained), while the other isoprostanes were largely ineffective. In the porcine arteries, on the other hand, all three F-ring isoprostanes as well as 8-iso PGE2 evoked constrictor responses, although the peak magnitudes were approximately 50% of the KCl-evoked response; 8-iso PGE2 and 8-iso PGF2alpha were the most potent, with negative log EC50 values of 6.5. We next sought to characterize the signaling pathways underlying the vasoconstrictor responses to 8-iso PGE2, since this was the most potent of the isoprostanes we tested. These responses were largely reversed by the thromboxane A2-selective (TP) prostanoid receptor antagonist ICI 192605 (10-8 m; 4(Z)-6-[(2,4,5 cis)2-(2-chlorophenyl)-4-(2-hydroxy phenyl)1,3-dioxan-5-yl]hexenoic acid) as well as by the nonspecific tyrosine kinase inhibitor genistein (10-5 and 10-4 m), and were reversed approximately 50% by the Rho-kinase inhibitor Y27632 (10-5 m; (+)-(R)-trans-4-(1-aminoethyl)-N-(pyridyl) cyclohexanecarboxamide dihydrochloride). We conclude, therefore, that 8-iso PGE2 constricts bronchial vasculature through the activation of TP receptors, which in turn trigger tyrosine kinase and Rho-kinase activities, resulting in powerful vasoconstriction. These findings are highly relevant to lung transplantation and to exercise-induced asthma.
Asunto(s)
Alprostadil/análogos & derivados , Arterias Bronquiales/efectos de los fármacos , Dinoprostona/análogos & derivados , Isoprostanos/farmacología , Músculo Liso Vascular/efectos de los fármacos , Porcinos , Vasoconstricción/efectos de los fármacos , Alprostadil/farmacología , Amidas/farmacología , Animales , Arterias Bronquiales/fisiología , Dinoprost/análogos & derivados , Dinoprost/farmacología , Dinoprostona/antagonistas & inhibidores , Dinoprostona/farmacología , Dioxanos/farmacología , Relación Dosis-Respuesta a Droga , F2-Isoprostanos/farmacología , Genisteína/farmacología , Humanos , Isoprostanos/antagonistas & inhibidores , Prostaglandinas E/farmacología , Prostaglandinas F Sintéticas/farmacología , Piridinas/farmacología , Receptores de Tromboxano A2 y Prostaglandina H2/efectos de los fármacos , Receptores de Tromboxano A2 y Prostaglandina H2/fisiología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Vasoconstricción/fisiología , Vasoconstrictores/antagonistas & inhibidores , Vasoconstrictores/farmacologíaRESUMEN
In the present experiment, we examined the effect of 8-iso-prostaglandin E(2) and 8-iso-prostaglandin F(2 alpha) on the release of noradrenaline from the isolated rat stomach. The postganglionic sympathetic nerves were electrically stimulated twice at 1 Hz for 1 min and test reagents were added during the second stimulation. 8-Iso-prostaglandin E(2) (10(-8)-10(-6) M) and 8-iso-prostaglandin F(2 alpha) (10(-7)-10(-5) M) dose-dependently reduced the evoked noradrenaline release, and these inhibitory potencies were as follows: 8-iso-prostaglandin E(2)>8-iso-prostaglandin F(2 alpha). The inhibitory effect of 8-iso-prostaglandin F(2 alpha), but not 8-iso-prostaglandin E(2), was abolished by 10(-6) M SQ-29548 ([1S-[1 alpha,2 alpha(Z),3 alpha,4 alpha]]-7-[3-[[2-[(phenylamino)carbonyl]hydrazino] methyl]-7-oxabicyclo[2,2,1]hept-2-yl]-5-heptenoic acid) (a prostanoid TP receptor antagonist). On the other hand, the inhibitory effect of 8-iso-prostaglandin E(2) was abolished by 10(-5) M AH-6809 (6-isopropoxy-9-oxoxanthene-2-carboxylic acid) (a prostanoid EP receptor antagonist), which also attenuated the inhibitory effects of ONO-AE-248 (16S-9-deoxy-9 beta-chloro-15-deoxy-16-hydroxy-17,17-trimethylene 19, 20-didehydro prostaglandin F(2)) (a selective EP(3) receptor agonist) on the evoked release of noradrenaline. The inhibitory effect of 8-iso-prostaglandin F(2 alpha), but not 8-iso-prostaglandin E(2), was abolished by pertussis toxin. These results suggest that 8-iso-prostaglandin F(2 alpha) inhibits noradrenaline release through TP receptors, whereas 8-iso-prostaglandin E(2) seems to inhibit noradrenaline release through EP(3) receptors, located on the gastric sympathetic nerve terminals in rats.
Asunto(s)
Dinoprost/análogos & derivados , Dinoprostona/análogos & derivados , Dinoprostona/farmacología , F2-Isoprostanos/farmacología , Isoprostanos/farmacología , Norepinefrina/metabolismo , Estómago/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Mucosa Gástrica/metabolismo , Masculino , Norepinefrina/antagonistas & inhibidores , Ratas , Ratas WistarAsunto(s)
Biomarcadores/análisis , Dinoprost/análogos & derivados , Isoprostanos/metabolismo , Peroxidación de Lípido/fisiología , F2-Isoprostanos/metabolismo , F2-Isoprostanos/farmacología , Humanos , Hipertensión/fisiopatología , Estrés Oxidativo , Activación Plaquetaria/efectos de los fármacos , Sistema Vasomotor/fisiologíaRESUMEN
Recent studies show that 8-iso-prostaglandin F(2alpha), a member of F(2)-isoprostane family, acts as a vasoconstrictor via TP receptor activation; and its local release may contribute to an abnormal vasomotor tone associated with hypercholesterolemia. The purpose of this study was to examine whether ramatroban, a TP receptor antagonist, improves abnormal vascular reactivity in vivo in hypercholesterolemic rabbits. The plasma 8-iso-prostaglandin F(2alpha) levels in hypercholesterolemic groups were significantly higher than those in normal groups. The treatment by ramatroban reversed the attenuation of the vascular response to acetylcholine in hypercholesterolemic groups. However, L-N(G)-nitroarginine methyl ester, a nitric oxide synthase inhibitor, did not inhibit the protective effects of ramatroban. Attenuation of the vascular response to acetylcholine in hypercholesterolemic rabbits was significantly enhanced by 8-iso-prostaglandin F(2alpha). Attenuation of the vascular response to acetylcholine by a cholesterol-rich diet and 8-iso-prostaglandin F(2alpha) was canceled by ramatroban. These findings suggest that ramatroban improves the vascular response in vivo to acetylcholine in hypercholesterolemic rabbits by blocking the action of 8-iso-prostaglandin F(2alpha).
Asunto(s)
Acetilcolina/farmacología , Carbazoles/farmacología , Dinoprost/análogos & derivados , Hipercolesterolemia/fisiopatología , Receptores de Tromboxanos/antagonistas & inhibidores , Sulfonamidas/farmacología , Resistencia Vascular/efectos de los fármacos , Animales , F2-Isoprostanos/sangre , F2-Isoprostanos/farmacología , Hipercolesterolemia/sangre , Masculino , Nitroprusiato/farmacología , Conejos , Vasoconstrictores/farmacología , Vasodilatadores/farmacologíaRESUMEN
BACKGROUND AND PURPOSE: Free radical-induced peroxidation is an important factor in the genesis of hypoxic-ischemic encephalopathy, including that of the preterm infant. Isoprostanes are major peroxidation products. Since microvascular dysfunction seems to contribute to ischemic encephalopathies, we studied the cytotoxicity of 8-iso-prostaglandin F2alpha (PGF2alpha) on cerebral microvascular cells. METHODS: Microvascular endothelial, astroglial, and smooth muscle cells from newborn brain were cultured. The cytotoxicity of 8-iso-PGF2alpha on these cells was determined by MTT assays and lactate dehydrogenase (LDH) release, propidium iodide incorporation, and DNA fragmentation (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling [TUNEL]). In addition, effects of intraventricular injections of 8-iso-PGF2alpha and possible involvement of thromboxane in 8-iso-PGF2alpha-induced cytotoxicity were determined. RESULTS: 8-Iso-PGF2alpha induced time- and concentration-dependent endothelial cell death (EC50=0.1 nmol/L) but exerted little effect on smooth muscle and astroglial cells; endothelial cell death seemed mostly of oncotic nature (propidium iodide incorporation and LDH release). Cell death was associated with increased endothelial thromboxane A2 (TXA2) formation and was prevented by TXA2 synthase inhibitors (CGS12970 and U63557A); TXA2 mimetics U46619 and I-BOP also caused endothelial cell death. Intraventricular injection of 8-iso-PGF2alpha induced periventricular damage, which was attenuated by CGS12970 pretreatment. CONCLUSIONS: These data disclose a novel action of 8-iso-PGF2alpha involving TXA2 in oxidant stress-induced cerebral microvascular injury and brain damage.
Asunto(s)
Isquemia Encefálica/metabolismo , Encéfalo/irrigación sanguínea , Dinoprost/análogos & derivados , Dinoprostona/análogos & derivados , Endotelio Vascular/efectos de los fármacos , F2-Isoprostanos/farmacología , Microcirculación/efectos de los fármacos , Animales , Astrocitos/citología , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/patología , Muerte Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Fragmentación del ADN/efectos de los fármacos , Dinoprostona/farmacología , Relación Dosis-Respuesta a Droga , Endotelio Vascular/citología , Endotelio Vascular/metabolismo , Inhibidores Enzimáticos/farmacología , Técnicas In Vitro , Inyecciones Intraventriculares , Isoprostanos/farmacología , L-Lactato Deshidrogenasa/metabolismo , Microcirculación/citología , Microcirculación/metabolismo , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Necrosis , Ratas , Ratas Sprague-Dawley , Porcinos , Tromboxano A2/metabolismo , Tromboxano-A Sintasa/antagonistas & inhibidoresRESUMEN
The aim of this study was to quantify lung oxidant stress after short-term ozone exposure as reflected by 8-isoprostane concentrations in exhaled breath condensate (EBC) and to investigate the effects of inhaled budesonide on this response. 8-Isoprostane is a prostaglandin-F(2 alpha) isomer that is formed in vivo by free radical-catalyzed peroxidation of arachidonic acid. EBC is a noninvasive method to collect airway secretions. We undertook a double-blind, randomized, placebo-controlled, crossover study with inhaled budesonide (800 microg) or placebo twice daily for 2 weeks prior to ozone exposure (400 parts per billion) for 2 h in nine healthy nonsmokers. Exhaled 8-isoprostane was measured by an enzyme immunoassay. 8-Isoprostane was increased 4 h after ozone exposure compared to pre-exposure values in both placebo (36.9 +/- 3.9 pg/ml, mean +/- SEM, vs. 16.9 +/- 0.7 pg/ml; p <.001) and budesonide groups (33.4 +/- 2.6 pg/ml vs. 15.8 +/- 0.3 pg/ml; p <.001). Pretreatment with budesonide did not affect the increases in 8-isoprostane (mean differences 3.4 pg/ml, 95% CI -8.9 to 15.7, p =.54). Short-term ozone exposure causes acute increase in lung oxidative stress as reflected by exhaled 8-isoprostane. This increase is resistant to pretreatment with a high dose of inhaled budesonide.
Asunto(s)
Budesonida/farmacología , Dinoprost/química , F2-Isoprostanos/farmacología , Estrés Oxidativo , Adulto , Pruebas Respiratorias , Estudios Cruzados , Método Doble Ciego , Femenino , Radicales Libres , Humanos , Técnicas para Inmunoenzimas , Pulmón/fisiología , Masculino , Ozono/farmacología , Placebos , Isoformas de Proteínas , Distribución Aleatoria , Fumar , Esputo/metabolismo , Factores de Tiempo , Vasoconstrictores/farmacologíaRESUMEN
BACKGROUND: Diabetic pregnancy displays increased incidence of congenital malformations and elevated levels of lipid peroxides in the offspring. The aim of the present work was to study if exogenous administration of one lipid peroxide, the isoprostane 8-iso-PGF(2alpha), is teratogenic per se in rat embryos in vitro, and if such teratological effects may be diminished by supplementation of an antioxidative agent, i.e., N-acetylcysteine or superoxide dismutase, to the culture medium. METHODS: Day-9 embryos were cultured in vitro for 48 hr and subjected to 8-iso-PGF(2alpha) with and without N-acetylcysteine or superoxide dismutase. RESULTS: Addition of 2 micromol/l of the isoprostane 8-iso-PGF(2alpha) to the culture medium caused high malformation rate, decreased protein and DNA contents, decreased somite number and crown-rump-length as well as marked accumulation of the isoprostane in the embryonic tissues. Adding N-acetylcysteine or superoxide dismutase to the culture medium with isoprostane normalized almost all morphological and biochemical parameters, including the elevated tissue concentration of 8-iso-PGF(2alpha). CONCLUSIONS: Results indicate that the isoprostane (8-iso-PGF(2alpha)) serves both as an oxidative stress indicator and a teratogenic agent. The findings support earlier studies of enhanced oxidative stress and increased malformation rate in embryos exposed to a diabetes-like environment, and suggest prevention of dysmorphogenesis by administration of antioxidative agents.
Asunto(s)
Dinoprost/análogos & derivados , Embrión de Mamíferos/efectos de los fármacos , F2-Isoprostanos/farmacología , Peroxidación de Lípido , Acetilcisteína/farmacología , Animales , Antioxidantes/farmacología , Medios de Cultivo/metabolismo , Depuradores de Radicales Libres/farmacología , Técnicas de Cultivo de Órganos , Estrés Oxidativo , Ratas , Ratas Sprague-Dawley , Somitos/metabolismo , Superóxido Dismutasa/farmacología , Teratógenos , Factores de Tiempo , Vasoconstrictores/farmacologíaRESUMEN
Hypercholesterolemia (HC) is characterized by increased circulating 8-epi-prostaglandin-F(2alpha) (isoprostane), a vasoconstrictor, marker, and mediator of increased oxidative stress, whose vascular effects might be augmented in HC. Anesthetized pigs were studied in vivo with electron beam computed tomography after a 12-wk normal (n = 8) or HC (n = 8) diet. Mean arterial pressure (MAP), single-kidney perfusion, and glomerular filtration rate (GFR) were quantified before and during unilateral intrarenal infusions of U46619 (10 ng x kg(-1) x min(-1)) or isoprostane (1 microg x kg(-1) x min(-1)). Basal renal perfusion and function were similar, and isoprostane infusion elevated its systemic levels similarly in normal and HC (333 +/- 89 vs. 366 +/- 48 pg/ml, respectively, P < 0.01 vs. baseline). Both drugs markedly and comparably decreased cortical perfusion and GFR in both groups, whereas medullary perfusion decreased significantly only in HC. Moreover, MAP increased significantly only in HC (+9 +/- 3 and +11 +/- 3 mmHg, respectively, PAsunto(s)
Dinoprost/análogos & derivados
, F2-Isoprostanos/farmacología
, Hipercolesterolemia/fisiopatología
, Riñón/efectos de los fármacos
, Riñón/fisiopatología
, Vasoconstrictores/farmacología
, Sistema Vasomotor/efectos de los fármacos
, Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología
, Animales
, Presión Sanguínea/efectos de los fármacos
, Relación Dosis-Respuesta a Droga
, Vías de Administración de Medicamentos
, Femenino
, Tasa de Filtración Glomerular/efectos de los fármacos
, Hemodinámica/efectos de los fármacos
, Isoprostanos/farmacología
, Riñón/irrigación sanguínea
, Flujo Sanguíneo Regional/efectos de los fármacos
, Porcinos
, Tomografía Computarizada por Rayos X
, Sistema Vasomotor/fisiopatología