RESUMEN
Genital tract bacterial infections could induce abortion and are some of the most common complications of pregnancy; however, the mechanisms remain unclear. We investigated the role of prostaglandins (PGs) in the mechanism of bacterial lipopolysaccharide (LPS)-induced pregnancy loss in a mouse model, and we hypothesized that PGs might play a central role in this action. LPS increased PG production in the uterus and decidua from early pregnant mice and stimulated cyclooxygenase (COX)-II mRNA and protein expression in the decidua but not in the uterus. We also observed that COX inhibitors prevented embryonic resorption (ER). To study the possible interaction between nitric oxide (NO) and PGs, we administered aminoguanidine, an inducible NO synthase inhibitor. NO inhibited basal PGE and PGF(2alpha) production in the decidua but activated their uterine synthesis and COX-II mRNA expression under septic conditions. A NO donor (S-nitroso-N-acetylpenicillamine) produced 100% ER and increased PG levels in the uterus and decidua. LPS-stimulated protein nitration was higher in the uterus than in the decidua. Quercetin, a peroxynitrite scavenger, did not reverse LPS-induced ER. Our results suggest that in a model of septic abortion characterized by increased PG levels, NO might nitrate and thus inhibit COX catalytic activity. ER prevention by COX inhibitors adds a possible clinical application to early pregnancy complications due to infections.
Asunto(s)
Reabsorción del Feto/inducido químicamente , Reabsorción del Feto/metabolismo , Lipopolisacáridos/farmacología , Óxido Nítrico/metabolismo , Prostaglandinas/metabolismo , Animales , Inhibidores de la Ciclooxigenasa/farmacología , Femenino , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico Sintasa/metabolismo , Embarazo , Prostaglandina-Endoperóxido Sintasas/genética , Prostaglandina-Endoperóxido Sintasas/metabolismo , ARN Mensajero/genética , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Tirosina/metabolismoRESUMEN
The production of prostaglandins (PGs) and nitric oxide (NO) by amnion tissue may play a significant role in parturition. It is thought that epidermal growth factor (EGF) may be one of the fetal signals that governs the initiation of labor. The aim of the present study was to investigate the effect of EGF in vivo on the PGs and nitrite production of rat fetal membranes. We have evaluated the regulation of PGs and nitrite production in rat fetal membranes ex vivo. The intra-uterine administration of EGF 500 ng in day 21 of pregnancy induced increases in PGE(2) (P<0.001) and PGF(2alpha) (P<0.01) compared to the control fetal membranes from pregnant rats on day 22. Also, this dose of EGF diminished nitrate production significantly (P<0.01). We found that fetal membranes at term (days 18-22 of gestation) expressed EGF-R. The NO donor, nitroprussiate 300 and 600 microM, elicited an inhibitory effect on the PGE(2) and PGF(2alpha) stimulated synthesis. On the other hand, indomethacin 10(-6) and 10(-7)M, a non-selective cyclooxygenase inhibitor, reverted the inhibitory effect exerted by EGF. Hence, rat fetal membranes were found to express epidermal growth factor receptors and, under the effect of EGF, PGs and nitrites production pathways interact probably to prevent a toxic effect caused by an exacerbated synthesis of these mediators.
Asunto(s)
Factor de Crecimiento Epidérmico/farmacología , Membranas Extraembrionarias/efectos de los fármacos , Membranas Extraembrionarias/metabolismo , Nitratos/metabolismo , Prostaglandinas/biosíntesis , Animales , Inhibidores de la Ciclooxigenasa/farmacología , Factor de Crecimiento Epidérmico/administración & dosificación , Receptores ErbB/metabolismo , Membranas Extraembrionarias/embriología , Femenino , Indometacina/farmacología , Isoenzimas/metabolismo , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico Sintasa/metabolismo , Nitroprusiato/farmacología , Embarazo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas WistarRESUMEN
Nitric oxide (NO) is synthesized in a variety of tissues, including rat uterus, from L-arginine by NO synthase (NOS), of which there are three isoforms, namely neuronal, endothelial and inducible NOS (nNOS, eNOS and iNOS, respectively). Nitric oxide is an important regulator of the biology and physiology of the organs of the reproductive system, including the uterus. Some studies have shown increased variation in NO production and NOS expression during the oestrous cycle. However, the factors that regulate NO production in the uterus remain unclear. Therefore, in the present study, we investigated the effect of sex steroids on NOS expression and activity in the ovariectomized rat uterus. Ovariectomized rats received progesterone (4 mg per rat) or 17beta-oestradiol (1 microg per rat). All rats were killed 18 h after treatment. Both progesterone and oestradiol were able to augment NOS activity. The effect of oestradiol was abolished by pre-incubation with 500 micro M aminoguanidine, an iNOS inhibitor, or by coadministration of oestradiol with 3 mg kg(-1) dexamethasone, but the effect of progesterone was not affected by these treatments. Uterine nNOS, eNOS and iNOS protein levels were assessed using Western blots. Ovariectomized rat uteri expressed iNOS and eNOS. Progesterone increased the expression of eNOS and iNOS, whereas oestradiol increased iNOS expression only. These results suggest that oestradiol and progesterone are involved in the regulation of NOS expression and activity during pregnancy and implantation in the rat.
Asunto(s)
Estradiol/farmacología , Hormonas Esteroides Gonadales/farmacología , Óxido Nítrico Sintasa/metabolismo , Progesterona/farmacología , Útero/efectos de los fármacos , Animales , Western Blotting , Calcio , Inhibidores Enzimáticos/farmacología , Estradiol/fisiología , Femenino , Hormonas Esteroides Gonadales/fisiología , Guanidinas/farmacología , Isoenzimas/análisis , Isoenzimas/antagonistas & inhibidores , Isoenzimas/metabolismo , Óxido Nítrico Sintasa/análisis , Óxido Nítrico Sintasa/antagonistas & inhibidores , Ovariectomía , Progesterona/fisiología , Ratas , Ratas Wistar , Regulación hacia Arriba , Útero/enzimologíaRESUMEN
1. The therapeutic effect of nonsteroidal anti-inflammatory drugs (NSAIDs) is thought to be due mainly to its inhibition of cyclooxygenase (COX) enzymes, but there is a growing body of research that now demonstrates a variety of NSAIDs effects on cellular signal transduction pathways other than those involving prostaglandins. 2. Nitric oxide (NO) as a free radical and an agent that gives rise to highly toxic oxidants (peroxynitrile, nitric dioxide, nitron ion), becomes a cause of neuronal damage and death in some brain lesions such as Parkinson and Alzheimer disease, and Huntington's chorea. 3. In the present study, the in vivo effect of three NSAIDs (lysine clonixinate (LC), indomethacine (INDO) and meloxicam (MELO)) on NO production and nitric oxide synthase expression in rat cerebellar slices was analysed. Rats were treated with (a) saline, (b) lipopolysaccharide (LPS) (5 mg kg(-1), i.p.), (c) saline in combination with different doses of NSAIDs and (d) LPS in combination with different doses of NSAIDs and then killed 6 h after treatment. 4. NO synthesis, evaluated by Bred and Snyder technique, was increased by LPS. This augmentation was inhibited by coadministration of the three NSAIDs assayed. None of the NSAIDs tested was able to modify control NO synthesis. 5. Expression of iNOS and neural NOS (nNOS) was detected by Western blotting in control and LPS-treated rats. LC and INDO, but not MELO, were able to inhibit the expression of these enzymes. 6. Therefore, reduction of iNOS and nNOS levels in cerebellum may explain, in part, the anti-inflammatory effect of these NSAIDs and may also have importance in the prevention of NO-mediated neuronal injury.
Asunto(s)
Cerebelo/efectos de los fármacos , Cerebelo/enzimología , Inhibidores de la Ciclooxigenasa/farmacología , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico/biosíntesis , Animales , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/fisiología , Lipopolisacáridos/farmacología , Masculino , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo I , Óxido Nítrico Sintasa de Tipo II , Ratas , Ratas WistarRESUMEN
Nitric oxide (NO) fulfils important functions during pregnancy and has a role in implantation, decidualization, vasodilatation and myometrial relaxation. However, at high concentrations, such as those that are produced in sepsis, NO has toxic effects as it is a free radical. The aim of this study was to characterize uterine and decidual NO production in lipopolysaccharide (LPS)-induced embryonic resorption in mice and to determine which isoforms of nitric oxide synthase (NOS) take part. LPS produced 100% embryonic resorption at 24 h, with complete fetus expulsions at 48 h. Decidual and uterine NO production were increased by LPS, with maximum production at 6 h. This increase was due to the induction of expression of inducible nitric oxide synthase (iNOS) isoform in the decidua and uterus, and neuronal nitric oxide synthase (nNOS) isoform in the decidua, as detected by western blot analysis and immunohistochemistry. LPS increased iNOS expression in decidual and myometrial cells and increased nNOS expression in decidual cells. In addition, LPS caused fibrinolysis and infiltration of mesometrial decidua by macrophages positive for iNOS and CD14 (LPS receptor). Endothelial nitric oxide synthase (eNOS) was found in decidual and uterine arteries but LPS did not modify its expression. LPS induced CD14 expression in endometrial glands, and this could have amplified the inflammatory response. Aminoguanidine, an inhibitor of iNOS activity, totally reversed the LPS-induced embryonic resorption. This result could be explained by an inhibition of the increase in NO production but also by an inhibition of the cellular infiltration and fibrinolysis. These results show that NO fulfils a fundamental role in LPS-induced embryonic resorption.
Asunto(s)
Infecciones Bacterianas/metabolismo , Pérdida del Embrión/metabolismo , Óxido Nítrico/metabolismo , Complicaciones Infecciosas del Embarazo/metabolismo , Animales , Infecciones Bacterianas/inmunología , Western Blotting/métodos , Decidua/enzimología , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Femenino , Fibrinólisis , Guanidinas/farmacología , Inmunohistoquímica/métodos , Receptores de Lipopolisacáridos/análisis , Lipopolisacáridos/farmacología , Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , Nitratos/análisis , Óxido Nítrico Sintasa/análisis , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II , Nitritos/análisis , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Útero/enzimologíaRESUMEN
Nitric oxide (NO) affects cyclooxygenase (COX) and lipooxygenase (LOX) activities in several tissues. The aim of this study was to investigate the effect of NO on the AA metabolism in the anterior pituitary. LOX and COX products from anterior pituitaries of Wistar male rats were determined by [14C]-AA radioconversion method. Sodium nitroprusside (NP, 0.5 mM) and DETA NONOate (1 mM), NO donors, decreased 5-hydroxy-5,8,11,14-eicosatetraenoic acid (5-HETE) synthesis (P<0.05), effects that were reversed by hemoglobin. L-arginine also inhibited LOX activity. To the contrary, the inhibition of NO synthase by L-NAME (0.5 mM) or aminoguanidine (0.5 mM) increased 5-HETE production (P<0.05). COX activity was slightly stimulated by NP and L-arginine. However, DETA NONOate induced a stimulation of the synthesis of all prostanoids (P<0.05), this effect being reversed by hemoglobin. Neither NOS inhibitors nor hemoglobin modified basal prostanoids synthesis. These results indicate that NO inhibits LOX activity and stimulates COX activity in the anterior pituitary gland. The inhibition of LOX by NO may be another mechanism involved in the effects of NO on hormone release in the anterior pituitary.
Asunto(s)
Ácido Araquidónico/metabolismo , Óxido Nítrico/fisiología , Adenohipófisis/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Depuradores de Radicales Libres/farmacología , Ácidos Hidroxieicosatetraenoicos/biosíntesis , Lipooxigenasa/efectos de los fármacos , Lipooxigenasa/metabolismo , Masculino , Óxido Nítrico/farmacología , Prostaglandina-Endoperóxido Sintasas/efectos de los fármacos , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas WistarRESUMEN
We have studied the effect of nitric oxide (NO) on the production of arachidonic acid ([14C]-AA) metabolites in the rat oviduct. The basal synthesis of eicosanoids was measured by the conversion of ([14C]-AA) to the different radiolabeled products of cyclooxygenase (COX). The oviducts incubated for 1 h with the labeled substrate of COX were able to convert 3.3 +/- 0.3% of ([14C]-AA) to 6-ceto-PGF1alpha, 10.7 +/- 1.0% to PGF2alpha, 13.5 +/- 1.2% to PGE2 and 6.3 +/- 0.5% to TXB2. The tissues were incubated with different doses of two NO donors: SIN-1 and Spermine NONOate. The results indicated that SIN-1 produces a significant decrease (50%; P < 0.05) in all prostanoids evaluated in a dose-response fashion. The inhibitory effect was completely reversed by addition of 20 microg/ml of hemoglobin (Hb), a NO scavenger. The addition of Spermine NONOate to the incubation medium diminished significantly (65%) the synthesis of COX metabolites suggesting that NO acts by inhibiting COX activity in the rat oviduct. However, NOS inhibitors, N(G)-L-arginine-methyl-ester (L-NAME) nd N(G)-L-monomethyl-arginine (L-NMMA) had no effect on basal production of the prostanoids. These results indicate that in the rat oviduct the synthesis of COX metabolites is negatively regulated by nitric oxide.
Asunto(s)
Ácido Araquidónico/metabolismo , Trompas Uterinas/metabolismo , Molsidomina/análogos & derivados , Óxido Nítrico/metabolismo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Prostaglandinas/biosíntesis , Espermina/análogos & derivados , Animales , Radioisótopos de Carbono/metabolismo , Trompas Uterinas/efectos de los fármacos , Femenino , Técnicas In Vitro , Molsidomina/farmacología , Óxido Nítrico/farmacología , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxidos de Nitrógeno , Ratas , Ratas Wistar , Espermina/farmacologíaRESUMEN
This study aims to examine the possible relationship between cytokines, nitric oxide (NO) and prostaglandins in the pregnant rat uterus. Results indicate that 1) IL1alpha enhances the synthesis of prostaglandins and augments NO production in pregnant rat uteri and 2) the effect of IL1alpha on prostaglandin synthesis is abolished by NMMA, a NOS inhibitor, by aminoguanidine, an iNOS inhibitor, and by NS-398, a COX-2 inhibitor. These results suggest that there is an interaction between IL1alpha, NO and prostaglandins and that are involved COX-2 and iNOS in this interrelationship. This mechanism might be important in the regulation of uterine contractility during pregnancy and labor.
Asunto(s)
Interleucina-1/farmacología , Interleucina-2/farmacología , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/metabolismo , Prostaglandinas/biosíntesis , Útero/metabolismo , Animales , Ácido Araquidónico/metabolismo , Radioisótopos de Carbono/metabolismo , Inhibidores de la Ciclooxigenasa/farmacología , Inhibidores Enzimáticos/farmacología , Femenino , Guanidinas/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Nitrobencenos/farmacología , Embarazo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas Wistar , Sulfonamidas/farmacología , Útero/efectos de los fármacos , omega-N-Metilarginina/farmacologíaRESUMEN
We examined the possible relationship between cytokines, nitric oxide and prostaglandins (PGs) in the estrogenized rat uterus. Results indicate that epidermal growth factor (EGF) enhances the synthesis of prostaglandins in estrogenized rat uteri and induces the augmentation of nitric oxide (NO) production in this tissue by stimulating iNOS. While the effect of EGF is abolished by L-NMMA, an NO antagonist, the NS-398, a cyclooxygenase-II (COX-II) inhibitor, prevents the augmentation of prostanoids induced by EGF. These results suggest that there is an interaction among EGF, NO and PGs and that in this interrelationship are involved COX-II and iNOS. This mechanism might be important during implantation and labor.
Asunto(s)
Factor de Crecimiento Epidérmico/farmacología , Estradiol/farmacología , Óxido Nítrico/metabolismo , Prostaglandinas/biosíntesis , Útero/metabolismo , Animales , Ácido Araquidónico/metabolismo , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa/farmacología , Femenino , Técnicas In Vitro , Isoenzimas/análisis , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Nitrobencenos/farmacología , Prostaglandina-Endoperóxido Sintasas/análisis , Ratas , Ratas Wistar , Sulfonamidas/farmacología , Útero/efectos de los fármacos , Útero/enzimología , omega-N-Metilarginina/farmacologíaRESUMEN
We examined the possible relationship between cytokines, nitric oxide (NO) and prostaglandins in the oestrogenized rat uterus. Results indicate that: IL-1alpha but not IL-2 enhances the synthesis of prostaglandins in oestrogenized rat uteri; IL-1alpha but not IL-2 induced an augmention of NO production in this tissue; the effect of IL-1alpha on prostaglandin synthesis is abolished by NMMA, an NO antagonist; NS-398, a COX-2 inhibitor, prevents the augmention of prostaglandins produced by IL-1alpha. These results suggest that there is an interaction between IL-1alpha, NO and prostaglandins and that this interrelationship involves COX-2. This mechanism might be important during implantation and labor.