RESUMEN
OBJECTIVE: To investigate the potential intraocular pressure (IOP)-lowering effects of nitric oxide (NO)-donating compounds in healthy canine eyes METHODS: A total of 79 dogs were divided into 3 groups in a masked, controlled and randomised study. Group N (n = 26) was administered 0.03% nitroglycerin in one eye and vehicle-control in the other, Group H (n = 26) was administered 0.1% hydralazine in one eye and vehicle-control in the other, while Group C (n = 27) received vehicle-control in both eyes (control group). Following eye drop administration, IOP was measured in both eyes at selected times (10-250 min), along with monitoring of heart rate and signs of ocular discomfort. Data was analysed with repeated measures mixed model and one-way ANOVA RESULTS: IOP was significantly reduced over the 4-h period with 0.03% nitroglycerin (p < 0.0001) but not 0.1% hydralazine (p = 0.520) when compared to contralateral vehicle-controlled eyes. IOP was reduced by up to 12% with 0.03% nitroglycerin from 10 to 70 min post-treatment; however, differences in IOP at individual time points were not statistically significant for either drug (p ≥ 0.133) as compared to contralateral vehicle-control eyes. No treatment group significantly affected heart rate (compared to Group C), and both treatment groups appeared well tolerated CONCLUSIONS: Both compounds were well-tolerated in healthy dogs. Nitroglycerin mildly reduced IOP in canine eyes, and further investigations are warranted in healthy and diseased states (e.g. glaucoma, ocular hypertension).
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Presión Intraocular , Nitroglicerina , Perros , Animales , Nitroglicerina/farmacología , Donantes de Óxido Nítrico/farmacología , Tonometría Ocular , Hidralazina/farmacologíaRESUMEN
OBJECTIVE: To describe the clinical application and effect of MicroPulse™ transscleral cyclophotocoagulation (MP-TSCPC) in dogs with glaucoma. ANIMALS STUDIED: Twelve dogs with primary (n = 8) or secondary (n = 4) glaucoma, aged 2-13 years (mean ± SD, 7.2 ± 3.8 years). PROCEDURES: MP-TSCPC was performed under sedation or general anesthesia. Laser duty cycle was 31.3%, laser power varied from 2000-2800 mW, and each hemisphere was treated for 90-180 seconds. The probe was applied to each quadrant in a "sweeping motion," sparing the 3 and 9 o'clock positions. RESULTS: The number of MP-TSCPC procedures per eye varied from 1 to 3 (1.4 ± 0.7). Intraocular pressure (IOP) was controlled (<25 mm Hg) in 11/12 dogs (92%) within 1-15 days post-operatively. The IOP control at 1 month and the duration between repeated procedures were significantly greater in eyes treated with high energy laser (2800 mW) compared to 2000-2500 mW. Long-term follow-up (315.3 ± 100.7 days) showed controlled IOP in 5/12 (42%) and vision retention in 4/8 (50%) dogs. In unsuccessful cases, loss of IOP control or vision loss occurred within 3-245 days (109.1 ± 93.7 days) and 28-261 days (114 ± 101.6 days), respectively, resulting in a salvage procedure in 6 dogs. Complications were as follows: corneal hypoesthesia (92%), anterior uveitis (67%), post-operative ocular hypertension (50%), neurotrophic corneal ulcer (25%), keratoconjunctivitis sicca (8%), and rubeosis iridis (8%). CONCLUSIONS: MP-TSCPC is a viable tool for managing canine glaucoma, although further studies are required to improve the long-term effect and reduce the complication rate.
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Glaucoma/veterinaria , Coagulación con Láser/veterinaria , Animales , Perros , Femenino , Glaucoma/cirugía , Presión Intraocular , Masculino , Cuidados Posoperatorios/veterinaria , Complicaciones Posoperatorias/veterinariaRESUMEN
OBJECTIVE: To evaluate the effect of topical ophthalmic 0.005% latanoprost alone and in combination with 0.1% diclofenac on healthy horses. ANIMALS STUDIED: Twelve healthy adult horses. PROCEDURES: A randomized, masked crossover design was used with horses divided into three groups for once daily treatment in one randomly selected eye. For arm 1 of the study, Group D (n = 3) received 0.1% diclofenac, Group L (n = 3) received 0.005% latanoprost, and Group DL (n = 6) received 0.005% latanoprost and 0.2 ml of 0.1% diclofenac. For arm 2 of the study, horses from Group D and L were placed into Group DL and horses from Group DL were placed into either Group D or L. Evaluations of intraocular pressure (IOP), vertical pupil diameter, aqueous flare, conjunctival hyperemia, epiphora, blepharospasm, and blepharoedema were performed 4 times daily on days 1 and 2 (baseline), days 3 to 7 (arm 1 treatment), days 8 to 11 (washout), days 12 to 16 (arm 2 treatment), and days 17 and 18 (return to baseline). RESULTS: During the treatment period, significant reduction in IOP and vertical pupil diameter occurred in treated eyes of Groups L and DL, but not Group D. These variables did not differ significantly between Groups L and DL. Blepharospasm, blepharoedema, epiphora, and conjunctival hyperemia scores were significantly higher in Group L than in Groups D and DL. CONCLUSIONS: Latanoprost reduced IOP in healthy horses and signs of drug-induced discomfort were mitigated by concurrent use of diclofenac.
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Antihipertensivos/farmacología , Diclofenaco/farmacología , Presión Intraocular/efectos de los fármacos , Soluciones Oftálmicas/farmacología , Prostaglandinas F Sintéticas/farmacología , Administración Tópica , Animales , Antihipertensivos/administración & dosificación , Diclofenaco/administración & dosificación , Método Doble Ciego , Quimioterapia Combinada/veterinaria , Caballos , Latanoprost , Soluciones Oftálmicas/administración & dosificación , Proyectos Piloto , Prostaglandinas F Sintéticas/administración & dosificación , Pupila/efectos de los fármacosRESUMEN
PURPOSE: To present the clinicopathologic features of a Domestic Short-haired cat with spontaneous, intermediate-grade corneal fibrosarcoma, possibly secondary to chronic corneal irritation associated with a corneal sequestrum. METHODS: A 12-year-old, spayed female Domestic Short-haired cat was evaluated for a slowly growing, pink, exophytic mass affecting the left cornea. The cat had presented 6 years previously for bilateral brown corneal sequestra, as well as 3 years previously for a small pale growth on the left cornea hypothesized to be an epithelial inclusion cyst and a corneal ulcer affecting the right eye. Incisional biopsy of the corneal mass indicated intermediate-grade corneal fibrosarcoma within the corneal stroma. Owing to the potential for malignant behavior, the left globe was enucleated. Routine systemic staging was performed prior to surgery with no evidence of metastasis. RESULTS: Definitive diagnosis of corneal fibrosarcoma was made through histopathologic examination of the incisional biopsy. There was an elevated mitotic index, indicating an intermediate-grade phenotype. Histopathology of the enucleated globe substantiated the initial findings, and complete tumor resection was confirmed. Subjacent to the corneal fibrosarcoma, there was a region of necrotic tissue suggestive of a corneal sequestrum. Six months after diagnosis and enucleation, the patient remained healthy with no signs of local spread or distant metastasis. CONCLUSIONS: To the authors' knowledge, this is the first documented case of a corneal fibrosarcoma in a cat.
RESUMEN
Glaucoma is a painful and often blinding group of ocular diseases for which there is no cure. Although the definition of glaucoma is rapidly evolving, elevated intraocular pressure (IOP) remains the most consistent risk factor of glaucoma in the canine patient. Therapy should be aimed at neuroprotection. The mainstay of therapy focuses on reducing IOP and maintaining a visual and comfortable eye. This article discusses the most current ocular hypotensive agents, focusing on their basic pharmacology, efficacy at lowering IOP, and recommended use in the treatment of idiopathic canine glaucoma.
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Enfermedades de los Perros/tratamiento farmacológico , Glaucoma/veterinaria , Agonistas Adrenérgicos/administración & dosificación , Agonistas Adrenérgicos/uso terapéutico , Animales , Inhibidores de Anhidrasa Carbónica/administración & dosificación , Inhibidores de Anhidrasa Carbónica/uso terapéutico , Agonistas Colinérgicos/administración & dosificación , Agonistas Colinérgicos/uso terapéutico , Perros , Sistemas de Liberación de Medicamentos/veterinaria , Gasotransmisores/uso terapéutico , Glaucoma/tratamiento farmacológico , Mióticos/administración & dosificación , Mióticos/uso terapéutico , Prostaglandinas/administración & dosificación , Prostaglandinas/uso terapéutico , Quinasas Asociadas a rho/antagonistas & inhibidoresRESUMEN
Practitioners approach chemical ciliary body ablation (CBA) in cats with caution. In 1994, an academic letter proposed a potential link between intraocular gentamicin injections for glaucoma and the appearance of ocular tumors in cats (Veterinary and Comparative Ophthalmology, 4, 1994, 166). There is an historic perceived risk for the development of feline ocular post-traumatic sarcoma following gentamicin ciliary body ablation, and many clinicians refrain from chemical ablation in cats for this reason. A recent study discussed the possibility of a correlation between intravitreal gentamicin and tumor promotion in dogs (Veterinary Ophthalmology, 16, 2013, 159). We searched the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW) database for cases of cats diagnosed with ocular tumors following ciliary body ablation. Of eight cases with historic gentamicin injection, five had malignant tumors: three post-traumatic sarcomas and two melanomas.
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Enfermedades de los Gatos/patología , Cuerpo Ciliar/patología , Neoplasias del Ojo/veterinaria , Gentamicinas/efectos adversos , Gentamicinas/farmacología , Dolor/veterinaria , Animales , Enfermedades de los Gatos/etiología , Gatos , Neoplasias del Ojo/etiología , Femenino , Glaucoma/tratamiento farmacológico , Glaucoma/veterinaria , Inyecciones Intravítreas/veterinaria , Masculino , Dolor/tratamiento farmacológico , Estudios RetrospectivosRESUMEN
Iridociliary tumors are the second most common primary ocular tumor in dogs and are usually benign. A review of the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW) database in 2009 suggested a potential correlation between malignant iridociliary epithelial tumors and ciliary body ablation by intravitreal gentamicin injection for the treatment of glaucoma. The purpose of this case series was to determine whether there is evidence of such a correlation in the COPLOW collection. Mining of the COPLOW database revealed that a significant number (39.5%) of canine globes with a history of ciliary body ablation were subsequently diagnosed with primary ocular tumors at enucleation, most commonly iridociliary epithelial tumors and melanocytic tumors. It is possible that neoplasia was present but unrecognized at the time of ciliary body ablation. These tumors had a higher than expected incidence of malignancy. These cases underscore the importance of reserving ciliary body ablation with gentamicin for disease-free eyes.
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Cuerpo Ciliar/efectos de los fármacos , Enfermedades de los Perros/inducido químicamente , Gentamicinas/farmacología , Neoplasias del Iris/veterinaria , Inhibidores de la Síntesis de la Proteína/farmacología , Animales , Perros , Gentamicinas/administración & dosificación , Glaucoma/tratamiento farmacológico , Glaucoma/veterinaria , Inyecciones Intravítreas/veterinaria , Neoplasias del Iris/inducido químicamente , Neoplasias del Iris/patología , Inhibidores de la Síntesis de la Proteína/administración & dosificación , Estudios RetrospectivosRESUMEN
UNLABELLED: What's known on the subject? and What does the study add? With the present study, we aimed to provide a global picture of the molecular processes that are activated by CN injury. The present study used genomic expression profiling to identify candidate genes that might be useful targets in the CN recovery process and, thus, the ultimate preservation of penile erection. Regeneration of the CN and axonal outgrowth clearly involve changes in multiple biochemical pathways that have never been investigated by microarray analysis. We analyzed global gene expression in the major pelvic ganglion at early stages (48 h and 14 days) after CN injury and focused on the detection of changes in genes related to nervous tissue repair and proliferation. The findings of the present study provide important insight into the molecular systems affected by CN injury and identify candidate genes that may be utilized for novel molecular-based therapies for the preservation and protection of the CN during RP. OBJECTIVES: To to examine the complexity of the many molecular systems involved in supporting cavernous nerve (CN) repair and regeneration in a rat model of bilateral crush injury utilizing a microarray analysis approach. Erectile dysfunction (ED) is a common clinical complication after prostate cancer treatment by radical prostatectomy, and recovery of erectile function can take as long as 2 years. There are gaps in our understanding of the autonomic pelvic innervation of the penis that still need to be addressed for the development of an adequate treatment strategy for post-prostatectomy ED. The molecular mechanisms of the intrinsic ability of CN to regenerate after an injury have not been elucidated. MATERIALS AND METHODS: We analyzed global gene expression in the major pelvic ganglion 48 h and 14 days after CN injury. Overall, a comparative analysis showed that 325 genes changed at the 48-h time point and 114 genes changed at 14 days. There were 60 changed genes in common with both time points. Using the Ingenuity Pathway Analysis® system (Ingenuity Systems, Inc., Redwood City, CA, USA), we were able to analyze the significantly changed genes that were unique and common to each time point by biological function. We focused on the detection of changes related to nervous tissue repair and proliferation, molecular networks of neurotrophic factors, stem cell regulation and synaptic transmission. RESULTS: There was strong evidence of the early mobilization of genes involved in repair and neuroprotection mechanisms (SERPINF1, IGF1, PLAU/PLAUR, ARG1). Genes related to nervous system development (ATF3 GJA1, PLAU, SERPINE1), nerve regeneration (SERPINE2, IGF1, ATF3, ARG1) and synaptic transmission (GJC1, GAL) were changed. Several genes related to proliferation as well as apoptosis (A2M, ATF3, C3, EGR4, FN1, GJA1, GAL) were also changed, possibly as part of a protective mechanism or the initiation of remodelling. CONCLUSIONS: The results obtained show that multiple biological processes are associated with injury and repair of the CN and provide a systematic genome-wide screen for neurotrophic and/or inhibitory pathways of nerve regeneration. These data identify the candidate genes that may be utilized in novel molecular-based therapies for the preservation and protection of the CN during radical prostatectomy.
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Disfunción Eréctil/genética , Ganglios/fisiopatología , Plexo Hipogástrico/fisiopatología , Regeneración Nerviosa/genética , Pene/inervación , ARN/análisis , Recuperación de la Función , Animales , Biomarcadores/metabolismo , Modelos Animales de Enfermedad , Disfunción Eréctil/etiología , Disfunción Eréctil/metabolismo , Disfunción Eréctil/fisiopatología , Ganglios/lesiones , Ganglios/metabolismo , Plexo Hipogástrico/lesiones , Plexo Hipogástrico/metabolismo , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Erección Peniana , Pene/lesiones , Pene/metabolismo , Ratas , Ratas Sprague-Dawley , Traumatismos del Sistema Nervioso/complicaciones , Traumatismos del Sistema Nervioso/metabolismo , Traumatismos del Sistema Nervioso/fisiopatologíaRESUMEN
AIMS: It has been well demonstrated that phosphodiesterase-5A (PDE5A) is expressed in smooth muscle cells and plays an important role in regulation of vascular tone. The role of endothelial PDE5A, however, has not been yet characterized. The present study was undertaken to determine the presence, localization, and potential physiologic significance of PDE5A within vascular endothelial cells. METHODS AND RESULTS: We demonstrate primary location of human, mouse, and bovine endothelial PDE5A at or near caveolae. We found that the spatial localization of PDE5A at the level of caveolin-rich lipid rafts allows for a feedback loop between endothelial PDE5A and nitric oxide synthase (NOS3). Treatment of human endothelium with PDE5A inhibitors resulted in a significant increase in NOS3 activity, whereas overexpression of PDE5A using an adenoviral vector, both in vivo and in cell culture, resulted in decreased NOS3 activity and endothelium-dependent vasodilation. The molecular mechanism responsible for these interactions is primarily regulated by cGMP-dependent second messenger. PDE5A overexpression also resulted in a significant decrease in protein kinase 1 (PKG1) activity. Overexpression of PKG1 rapidly activated NOS3, whereas silencing of the PKG1 gene with siRNA inhibited both NOS3 phosphorylation (S1179) and activity, indicating a novel role for PKG1 in direct regulation of NOS3. CONCLUSION: Our data collectively suggest another target for PDE5A inhibition in endothelial dysfunction and provide another physiologic significance for PDE5A in the modulation of endothelial-dependent flow-mediated vasodilation. Using both in vitro and in vivo models, as well as human data, we show that inhibition of endothelial PDE5A improves endothelial function.
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Caveolas/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 5/metabolismo , Células Endoteliales/enzimología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Vasodilatación/fisiología , Animales , Aorta/citología , Aorta/enzimología , Bovinos , Células Cultivadas , Vasos Coronarios/citología , Vasos Coronarios/enzimología , Proteína Quinasa Dependiente de GMP Cíclico Tipo I , Proteínas Quinasas Dependientes de GMP Cíclico/genética , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 5/genética , Células Endoteliales/citología , Humanos , Hipertensión Pulmonar/metabolismo , Hipertensión Pulmonar/patología , Microdominios de Membrana/metabolismo , Ratones , Arteria Pulmonar/citología , Arteria Pulmonar/enzimología , Transducción de Señal/fisiología , Venas Umbilicales/citología , Venas Umbilicales/enzimologíaRESUMEN
PURPOSE: RhoA and rho kinase serve as key regulators of penile vascular homeostasis. The role of RhoA/rho kinase signaling in the penis after cavernous nerve injury has not been fully investigated. We characterized the molecular expression profiles of RhoA/rho kinase signaling that occur in the penis after cavernous nerve injury. We hypothesized that erectile dysfunction after bilateral cavernous nerve injury is accompanied by up-regulation of RhoA/rho kinase activity in the rat penis. MATERIAL AND METHODS: We used 2 groups, including sham operation and bilateral cavernous nerve injury. At 14 days after nerve injury each group underwent cavernous nerve stimulation to determine erectile function at baseline and after intracavernous injection of the rho kinase inhibitor Y-27632 (Tocris Bioscience, Ellisville, Missouri). Penes were assessed at baseline for protein expression of neuronal nitric oxide synthase, RhoA, and rho kinase 1 and 2 by Western blot, immunoreactivity of neuronal nitric oxide synthase, rho kinase 1 and 2, RhoA-guanosine triphosphatase and rho kinase activity. RESULTS: Erectile function was decreased in nerve injured rats. Neuronal nitric oxide synthase protein was significantly decreased while RhoA and rho kinase 2 protein levels were significantly increased in rat penes with nerve injury. Rho kinase 1 protein expression was equivalent. Rho kinase immunoreactivity was qualitatively increased in the corporeal smooth muscle of nerve injured rats. RhoA-guanosine triphosphatase and rho kinase activity was significantly increased in injured rat penes compared to that in sham operated penes. Intracavernous injection of Y-27632 caused a significantly greater increase in intracavernous pressure in nerve injured rats compared to that in sham operated rats, suggesting increased rho kinase activity. CONCLUSIONS: Data suggest that RhoA/rho kinase up-regulation in response to cavernous nerve injury contributes to penile vasculature dysfunction after cavernous nerve injury. Thus, the RhoA/rho kinase pathway may be a suitable target for treating post-radical prostatectomy erectile dysfunction.
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Disfunción Eréctil/enzimología , Disfunción Eréctil/etiología , Pene/lesiones , Pene/inervación , Quinasas Asociadas a rho/fisiología , Animales , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
INTRODUCTION: Hypercholesterolemia induces erectile dysfunction (ED) mostly by increasing oxidative stress and impairing endothelial function in the penis, but the mechanisms regulating reactive oxygen species (ROS) production in the penis are not understood. AIMS: We evaluated whether hypercholesterolemia activates nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase in the penis, providing an initial source of ROS to induce endothelial nitric oxide synthase (eNOS) uncoupling and endothelial dysfunction resulting in ED. METHODS: Low-density-lipoprotein receptor (LDLR)-null mice were fed Western diet for 4 weeks to induce early-stage hyperlipidemia. Wild type (WT) mice fed regular chow served as controls. Mice received NAD(P)H oxidase inhibitor apocynin (10 mM in drinking water) or vehicle. Erectile function was assessed in response to cavernous nerve electrical stimulation. Markers of endothelial function (phospho [P]-vasodilator-stimulated-protein [VASP]-Ser-239), oxidative stress (4-hydroxy-2-nonenal [HNE]), sources of ROS (eNOS uncoupling and NAD[P]H oxidase subunits p67(phox) , p47(phox) , and gp91(phox) ), P-eNOS-Ser-1177, and eNOS were measured by Western blot in penes. MAIN OUTCOME MEASURES: The main outcome measures are the molecular mechanisms of ROS generation and endothelial dysfunction in hypercholesterolemia-induced ED. RESULTS: Erectile response was significantly (P<0.05) reduced in hypercholesterolemic LDLR-null mice compared with WT mice. Relative to WT mice, hypercholesterolemia increased (P<0.05) protein expressions of NAD(P)H oxidase subunits p67(phox) , p47(phox) and gp91(phox) , eNOS uncoupling, and 4-HNE-modified proteins, and reduced (P<0.05) P-VASP-Ser-239 expression in the penis. Apocynin treatment of LDLR-null mice preserved (P<0.05) maximal intracavernosal pressure, and reversed (P<0.05) the abnormalities in protein expressions of gp67(phox) and gp47(phox) , 4-HNE, P-VASP-Ser-239, and eNOS uncoupling in the penis. Apocynin treatment of WT mice did not affect any of these parameters. Protein expressions of P-eNOS-Ser-1177 and total eNOS were unaffected by hypercholesterolemia. CONCLUSION: Activated NAD(P)H oxidase in the penis is an initial source of oxidative stress resulting in eNOS uncoupling, thus providing a mechanism of eNOS uncoupling and endothelial dysfunction in hypercholesterolemia-induced ED.
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Endotelio Vascular/metabolismo , Disfunción Eréctil/metabolismo , Hipercolesterolemia/complicaciones , NADPH Oxidasas/metabolismo , Pene/metabolismo , Acetofenonas/farmacología , Aldehídos/metabolismo , Animales , Moléculas de Adhesión Celular/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Colesterol en la Dieta/administración & dosificación , Colesterol en la Dieta/efectos adversos , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Disfunción Eréctil/etiología , Masculino , Ratones , Proteínas de Microfilamentos/efectos de los fármacos , Proteínas de Microfilamentos/metabolismo , Óxido Nítrico Sintasa de Tipo III/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo III/metabolismo , Pene/inervación , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/metabolismo , Especies Reactivas de OxígenoRESUMEN
OBJECTIVES: The Ras homolog gene family, member A (RhoA) and its main downstream effector, Rho-kinase (ROCK) are important in maintaining the penis in the flaccid state. The pathophysiology of sickle cell disease-associated priapism is not well defined. We hypothesized that the RhoA/ROCK vasoconstrictive pathways might be involved in the development of priapism. Therefore, the objective of the present study was to evaluate the molecular changes in RhoA and ROCK in an established transgenic sickle cell mouse model of priapism. METHODS: Two groups of mice were used: wild type (WT; C57BL/6) mice and transgenic sickle cell mice. We evaluated RhoA guanosine triphosphatase and total ROCK activities, as well as ROCK1 and ROCK2 protein expression, in WT and sickle mice penises. We also evaluated the in vivo erectile responses to cavernous nerve stimulation and the frequency and duration of spontaneous erections before and after cavernous nerve stimulation. RESULTS: Sickle mice demonstrated significantly (P <.05) enhanced erectile responses to cavernous nerve stimulation and frequency of spontaneous erections both before and after cavernous nerve stimulation compared with the WT mice. The sickle mice penises had a significant decline in RhoA guanosine triphosphatase (P <.01) and total ROCK activities (P <.05) compared with the WT mice. A significant (P <.05) reduction in ROCK2 protein expression in sickle mice penises compared with WT mice protein expression. No change in ROCK1 protein expression was observed in either cohort of mice penises. CONCLUSIONS: These data suggest that sickle cell disease associated-priapism might be contributed by a lack of RhoA/ROCK-mediated vasoconstriction and highlight a novel molecular mechanism in the pathophysiology of priapism.
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Pene , Priapismo/enzimología , Priapismo/etiología , Quinasas Asociadas a rho/fisiología , Anemia de Células Falciformes/complicaciones , Anemia de Células Falciformes/enzimología , Animales , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Pene/irrigación sanguínea , Pene/enzimología , VasoconstricciónRESUMEN
INTRODUCTION: Estrogens control vaginal blood flow during female sexual arousal mostly through nitric oxide (NO). Although vascular effects of estrogens are attributed to an increase in endothelial NO production, the mechanisms of endothelial NO synthase (eNOS) regulation by estrogens in the vagina are largely unknown. AIMS: Our hypothesis was that estrogens regulate eNOS post-translationally in the vagina, providing a mechanism to affect NO bioavailability without changes in eNOS protein expression. METHODS: We measured eNOS phosphorylation and eNOS interaction with caveolin-1 and heat shock protein 90 (HSP90) in the distal and proximal vagina of female rats at diestrus, 7 days after ovariectomy and 2 days after replacement of ovariectomized rats with estradiol-17beta (15 microg). MAIN OUTCOME MEASURES: Molecular mechanisms of eNOS regulation by estrogen in the rat vagina. RESULTS: We localized phospho-eNOS (Ser-1177) immunohistochemically to the endothelium lining blood vessels and vaginal sinusoids. Estrogen withdrawal decreased phosphorylation of eNOS on its positive regulatory site (Ser-1177) and increased eNOS binding to its negative regulator caveolin-1 (without affecting eNOS/HSP90 interaction), and they were both normalized by estradiol replacement. Protein expressions of phosphorylated Akt (protein kinase B) and extracellular signal-regulated protein kinase 1/2 (ERK1/2) were not affected by estrogen status, suggesting that the effect of estrogens on eNOS (Ser-1177) phosphorylation was not mediated by activated AKT or ERK1/2. eNOS phosphorylation on its negative regulatory site (Ser-114) was increased in the vagina by estrogen withdrawal and normalized by estradiol replacement, implying that the maintenance of low phosphorylation of eNOS on this site by estradiol may limit eNOS interaction with caveolin-1 and preserve the enzyme's activity. Total eNOS, inducible NOS, caveolin-1, and HSP90 protein expressions were not affected by ovariectomy or estradiol replacement in the distal or proximal vagina. CONCLUSIONS: These results define novel estrogen signaling mechanisms in the vagina which involve eNOS phosphorylation and eNOS-caveolin-1 interaction.
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Estrógenos/fisiología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Vagina/metabolismo , Animales , Western Blotting , Caveolina 1/metabolismo , Estradiol/farmacología , Femenino , Proteínas HSP90 de Choque Térmico/metabolismo , Fosforilación , Procesamiento Proteico-Postraduccional , Ratas , Ratas Endogámicas F344 , Útero/efectos de los fármacos , Útero/metabolismo , Vagina/efectos de los fármacosRESUMEN
INTRODUCTION: Female sexual arousal disorder (FSAD) is a major component of female sexual dysfunctions, affecting 25-70% of women. The mechanisms of FSAD are poorly understood. Estrogen contributes to the control of genital blood flow during the sexual response. Vascular effects of estrogen are mostly attributed to its regulation of endothelial nitric oxide (NO) production. However, the role of endothelial NO synthase (eNOS) and the mechanisms that regulate eNOS in female genital tract structures are largely unknown. AIM: To review available evidence of the mechanisms of eNOS regulation in female genital tract structures. METHODS: This article reviews the literature that relates to the role of NO and eNOS in female sexual arousal and its modulation by estrogen. MAIN OUTCOME MEASURES: Association between female sexual arousal, NO, and eNOS. RESULTS: The NO/cyclic guanosine monophosphate pathway is believed to have a primary role in the regulation of clitoral and vaginal blood flow, and smooth muscle relaxation during sexual arousal. Estrogen is critical for maintaining vaginal and clitoral blood flow and vaginal transudate production. Estrogen regulates eNOS by genomic mechanisms, involving augmented mRNA transcription and protein synthesis, and by non-genomic mechanisms, which occur without alterations in gene expression. However, limited studies have evaluated the physiological role of endothelial NO and the molecular mechanisms of eNOS regulation in the female genital tract. CONCLUSIONS: The effects of estrogen on increasing genital blood flow and smooth muscle relaxation have been attributed mostly to regulation of eNOS. However, the exact mechanisms of eNOS regulation in female genital tract structures and the molecular basis for the eNOS defect with aging and vascular diseases warrant further investigation.
Asunto(s)
Endotelio Vascular/enzimología , Genitales Femeninos/enzimología , Óxido Nítrico Sintasa/metabolismo , Disfunciones Sexuales Fisiológicas/fisiopatología , Nivel de Alerta/fisiología , Caveolina 1/fisiología , Clítoris/irrigación sanguínea , Clítoris/metabolismo , GMP Cíclico/biosíntesis , Estrógenos/fisiología , Femenino , Humanos , Relajación Muscular/fisiología , Músculo Liso/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa/biosíntesis , ARN Mensajero/genética , Disfunciones Sexuales Fisiológicas/genética , Disfunciones Sexuales Fisiológicas/metabolismo , Vagina/irrigación sanguínea , Vagina/metabolismoRESUMEN
PURPOSE: We investigated changes in serum biomarkers of vascular function after short-term, continuous sildenafil dosing in men with type 2 diabetes with erectile dysfunction. MATERIALS AND METHODS: Men with erectile dysfunction associated with type 2 diabetes mellitus were randomized to receive continuous, daily sildenafil (50 mg for 1 week run-in and 100 mg for 3 weeks) (148), or placebo (144) for 4 weeks (phase I) and then sildenafil (25, 50 or 100 mg) on demand for 12 weeks (phase II). Blood draws at baseline and after phases I and II were analyzed for cyclic guanosine monophosphate (endothelial function marker), 8-isoprostane (oxidative stress marker), and interleukin-6 and interleukin-8 (inflammatory cytokines). Primary and secondary erectile function outcome variables were affirmative responses on Sexual Encounter Profile question 3 (ability to maintain erection sufficient for sexual intercourse) and Erection Hardness Score, respectively. RESULTS: Serum cyclic guanosine monophosphate levels were increased in the sildenafil group relative to the placebo group at 4 (p <0.01) and 16 (p <0.05) weeks, correlating with affirmative responses to Sexual Encounter Profile question 3 at the 4-week interval only (p <0.05). Serum 8-isoprostane levels were decreased to a nonsignificant degree in the sildenafil group at 4 weeks with no further change at 16 weeks, whereas interleukin-6 and interleukin-8 levels were unchanged at either interval, and these levels were unassociated with erectile function outcomes. CONCLUSIONS: These data suggest that short-term, continuous sildenafil treatment causes systemic endothelial function to be enhanced and remain so for a duration after its discontinuation. However, they do not indicate any influence of this treatment on systemic oxidative stress or inflammation, or an effect on long-term erectile function improvement.
Asunto(s)
GMP Cíclico/sangre , Complicaciones de la Diabetes/tratamiento farmacológico , Diabetes Mellitus Tipo 2/complicaciones , Dinoprost/análogos & derivados , Endotelio Vascular/efectos de los fármacos , Disfunción Eréctil/tratamiento farmacológico , Interleucina-6/sangre , Interleucina-8/sangre , Estrés Oxidativo/efectos de los fármacos , Inhibidores de Fosfodiesterasa/administración & dosificación , Piperazinas/administración & dosificación , Sulfonas/administración & dosificación , Adulto , Anciano , Biomarcadores/sangre , Dinoprost/sangre , Disfunción Eréctil/etiología , Humanos , Masculino , Persona de Mediana Edad , Inhibidores de Fosfodiesterasa/farmacología , Piperazinas/farmacología , Purinas/administración & dosificación , Purinas/farmacología , Citrato de Sildenafil , Sulfonas/farmacología , Resultado del TratamientoRESUMEN
PURPOSE: We determined the effect of passive secondhand cigarette smoke on 1) erectile function in vivo, 2) molecular mechanisms involved in penile vascular function, and 3) erectile function and penile molecular signaling in the presence of phosphodiesterase type 5 inhibitor therapy. MATERIALS AND METHODS: Four groups of mice were used, including group 1--controls, group 2--mice exposed to 3 weeks of secondhand smoke (5 hours per day for 5 days per week), group 3--control plus sildenafil (100 mg/kg per day) and group 4--smoke exposed plus sildenafil (100 mg/kg per day). Cavernous nerve electrical stimulation and intracavernous injection of acetylcholine were done to assess erectile function. Constitutive and inducible nitric oxide synthase activity, reactive oxygen species generation, nitrotyrosine formation and superoxide anion levels were assessed. RESULTS: Decreased erectile responses to cavernous nerve electrical stimulation and impaired endothelium dependent erectile responses to ACh in mice exposed to secondhand smoke were observed. Superoxide anion was increased in endothelial and corporeal smooth muscle cells of smoking mouse penises. In mice exposed to secondhand smoke constitutive nitric oxide synthase activity was decreased, and inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation increased. Sildenafil therapy restored constitutive nitric oxide synthase activity in the penis of smoking mice, decreased inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation, and improved erectile responses to cavernous nerve electrical stimulation and acetylcholine. CONCLUSIONS: Short-term exposure to secondhand smoke impairs erectile function through excessive penile reactive oxygen species signaling and inducible nitric oxide synthase activity. Decreased penile constitutive nitric oxide synthase activity may be attributable to the decreased endothelial nitric oxide synthase activity resulting from increased oxidative stress. Sildenafil therapy restored nitric oxide synthase activity and decreased reactive oxygen species signaling, resulting in improved erectile function.
Asunto(s)
Disfunción Eréctil/tratamiento farmacológico , Óxido Nítrico Sintasa/metabolismo , Inhibidores de Fosfodiesterasa/farmacología , Piperazinas/farmacología , Sulfonas/farmacología , Superóxidos/metabolismo , Administración Oral , Animales , Modelos Animales de Enfermedad , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Disfunción Eréctil/etiología , Disfunción Eréctil/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico Sintasa/efectos de los fármacos , Erección Peniana/efectos de los fármacos , Erección Peniana/fisiología , Purinas/farmacología , Distribución Aleatoria , Especies Reactivas de Oxígeno/metabolismo , Valores de Referencia , Sensibilidad y Especificidad , Transducción de Señal , Citrato de Sildenafil , Contaminación por Humo de Tabaco/efectos adversosRESUMEN
INTRODUCTION: The past 25 years of basic science research on erectile physiology has been devoted to investigating the pathogenesis of erectile dysfunction. Research has led to a better understanding of the biochemical factors and intracellular mechanisms responsible for corporal smooth muscle contraction and relaxation, as well as the influence of endothelial-derived relaxing factors. AIM: In this essay, we propose the use of gene transfer technology to study mechanisms of disease involved in penile vascular dysfunction. METHODS: The development of methods to deliver therapeutic genes to the penis has kindled a keen interest in treating ED with gene- and cell-based therapies. RESULTS: Gene therapy has delineated putative mechanisms of disease in animal models of erectile dysfunction. CONCLUSION: Investigation of animal models using gene therapy may ultimately lead to mechanism-based therapies for the treatment of erectile dysfunction.
Asunto(s)
Disfunción Eréctil/terapia , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Erección Peniana/genética , Animales , Disfunción Eréctil/genética , Humanos , Masculino , Modelos Animales , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/fisiología , Erección Peniana/fisiología , Flujo Sanguíneo RegionalRESUMEN
INTRODUCTION: Diet and exercise affect endothelial function in the penis, but the molecular mechanisms underlying their effects are not understood. AIMS: We evaluated endothelial nitric oxide synthase (eNOS) interaction with its negative regulator caveolin-1 and eNOS uncoupling as molecular targets in the penis associated with the beneficial effects of low-fat diet and chronic exercise. METHODS: The penes were obtained from adult male Yucatan pigs fed a normal-fat or high-fat diet on exercised or sedentary regimen for 24 weeks. Markers of endothelial function (guanosine 3',5'-monophosphate [cGMP] production), endothelial dysfunction (eNOS uncoupling and eNOS interaction with caveolin-1), and oxidative stress (thiobarbituric acid reactive substances [TBARS]) were measured in the penes. The concentrations of cGMP and TBARS were determined using commercial kits. eNOS uncoupling was determined by low-temperature sodium dodecyl sulfate polyacrylamide gel electrophoresis. eNOS binding to caveolin-1, eNOS phosphorylation (Ser-1177), and protein expression of eNOS and caveolin-1 were measured by Western blot analysis in penes purified for NOS and in homogenates, respectively. MAIN OUTCOME MEASURES: Molecular parameters of endothelial function including eNOS regulatory function. RESULTS: Relative to normal-fat diet, high-fat diet significantly (P < 0.05) reduced cGMP levels and significantly (P < 0.05) increased eNOS uncoupling, eNOS binding to caveolin-1, and TBARS production in the penis of sedentary pigs. Exercise of pigs on high-fat diet reversed (P < 0.05) the abnormalities in cGMP levels, eNOS uncoupling, and eNOS binding to caveolin-1, but not TBARS levels. Exercise of pigs on normal-fat diet did not affect any of these parameters. Protein expressions of caveolin-1, phosphorylated (Ser-1177), and total eNOS were unaffected by diet or exercise. CONCLUSION: Low-fat diet and chronic exercise preserve endothelial function in the pig penis by sustaining active eNOS in its dimeric form and by limiting eNOS interaction with its negative regulator caveolin-1.
Asunto(s)
Aterosclerosis/metabolismo , Dieta con Restricción de Grasas , Grasas de la Dieta , Óxido Nítrico Sintasa de Tipo III/metabolismo , Erección Peniana/fisiología , Condicionamiento Físico Animal/métodos , Animales , Western Blotting/métodos , Caveolina 1/metabolismo , GMP Cíclico/metabolismo , Masculino , Óxido Nítrico Sintasa de Tipo III/genética , Porcinos , Porcinos Enanos , Sustancias Reactivas al Ácido Tiobarbitúrico , Regulación hacia ArribaRESUMEN
Erectile dysfunction (ED) commonly results from endothelial dysfunction of the systemic vasculature. Although phosphodiesterase type 5 (PDE-5) inhibitors are effective at treating most cases of ED, they must be taken routinely and are ineffectual for a meaningful number of men. In recent years gene and stem cell-based therapies targeted at the penile endothelium have been gaining momentum in preclinical studies. These early studies reveal that gene and stem cell-based therapies may be both enduring and efficacious, and may eventually lead to a cure for ED. The following review will highlight our current understanding of endothelial-specific gene and stem cell-based therapies performed to date in a number of experimental animal models.