RESUMEN
Glaucoma is a blinding disease. Reduction of intraocular pressure (IOP) is the mainstay of treatment, but current drugs show side effects or become progressively ineffective, highlighting the need for novel compounds. We have synthesized a family of perhydro-1,4-oxazepine derivatives of digoxin, the selective inhibitor of Na,K-ATPase. The cyclobutyl derivative (DcB) displays strong selectivity for the human α2 isoform and potently reduces IOP in rabbits. These observations appeared consistent with a hypothesis that in ciliary epithelium DcB inhibits the α2 isoform of Na,K-ATPase, which is expressed strongly in nonpigmented cells, reducing aqueous humor (AH) inflow. This paper extends assessment of efficacy and mechanism of action of DcB using an ocular hypertensive nonhuman primate model (OHT-NHP) (Macaca fascicularis). In OHT-NHP, DcB potently lowers IOP, in both acute (24 h) and extended (7-10 days) settings, accompanied by increased aqueous humor flow rate (AFR). By contrast, ocular normotensive animals (ONT-NHP) are poorly responsive to DcB, if at all. The mechanism of action of DcB has been analyzed using isolated porcine ciliary epithelium and perfused enucleated eyes to study AH inflow and AH outflow facility, respectively. 1) DcB significantly stimulates AH inflow although prior addition of 8-Br-cAMP, which raises AH inflow, precludes additional effects of DcB. 2) DcB significantly increases AH outflow facility via the trabecular meshwork (TM). Taken together, the data indicate that the original hypothesis on the mechanism of action must be revised. In the OHT-NHP, and presumably other species, DcB lowers IOP by increasing AH outflow facility rather than by decreasing AH inflow.NEW & NOTEWORTHY When applied topically, a cyclobutyl derivative of digoxin (DcB) potently reduces intraocular pressure in an ocular hypertensive nonhuman primate model (Macaca fascicularis), associated with increased aqueous humor (AH) flow rate (AFR). The mechanism of action of DcB involves increased AH outflow facility as detected in enucleated perfused porcine eyes and, in parallel, increased (AH) inflow as detected in isolated porcine ciliary epithelium. DcB might have potential as a drug for the treatment of open-angle human glaucoma.
Asunto(s)
Humor Acuoso , Digoxina , Presión Intraocular , Macaca fascicularis , Hipertensión Ocular , Animales , Presión Intraocular/efectos de los fármacos , Digoxina/farmacología , Humor Acuoso/metabolismo , Humor Acuoso/efectos de los fármacos , Hipertensión Ocular/tratamiento farmacológico , Hipertensión Ocular/fisiopatología , Hipertensión Ocular/metabolismo , Modelos Animales de Enfermedad , Glaucoma/tratamiento farmacológico , Glaucoma/metabolismo , Glaucoma/fisiopatología , Conejos , Humanos , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , Masculino , Malla Trabecular/efectos de los fármacos , Malla Trabecular/metabolismoRESUMEN
This study investigated the effects of omidenepag (OMD), a novel selective EP2 receptor agonist, on human trabecular meshwork (HTM) cells, monkey Schlemm's canal endothelial (SCE) cells, and porcine ciliary muscle (CM) to clarify the mechanism of intraocular pressure (IOP) reduction involving conventional outflow pathway. In HTM and SCE cells, the effects of OMD on transforming growth factor-ß2 (TGF-ß2)-induced changes were examined. The expression of actin cytoskeleton and extracellular matrix (ECM) proteins, myosin light chain (MLC) phosphorylation in HTM cells were evaluated using real-time quantitative PCR, immunocytochemistry, and western blotting. The expression of barrier-related proteins, ZO-1 and ß-catenin, and permeability of SCE cells were evaluated using immunocytochemistry and transendothelial electrical resistance. The CM contraction was determined by contractibility assay. OMD significantly inhibited expression of TGF-ß2 induced mRNA, protein, and MLC-phosphorylation on cytoskeletal and ECM remodeling in the HTM dose dependently. In SCE cells, OMD suppressed TGF-ß2-induced expression of the barrier-related proteins and decreased SCE monolayer permeability. OMD at 3 µM significantly inhibited CM contraction, however, the effect was not significant at lower concentrations. IOP lowering effect of OMD through conventional outflow pathway is exerted by increasing outflow facilities with the modulation of TM cell fibrosis and SCE cell permeability.
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Cuerpo Ciliar/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Glicina/análogos & derivados , Contracción Muscular , Pirazoles/farmacología , Piridinas/farmacología , Subtipo EP2 de Receptores de Prostaglandina E/agonistas , Esclerótica/efectos de los fármacos , Malla Trabecular/efectos de los fármacos , Animales , Cuerpo Ciliar/metabolismo , Células Endoteliales/metabolismo , Glicina/farmacología , Humanos , Macaca fascicularis , Esclerótica/metabolismo , Porcinos , Malla Trabecular/metabolismoRESUMEN
Purpose: To investigate the effect of NKR-1 antagonists in an established UVR-B-induced cataract mouse model. Furthermore, to examine the expression of pro-inflammatory cytokines/chemokines in mouse eyes following unilateral UVR-B exposure.Methods: Mice received intraperitoneally injections of Fosaprepitant and Spantide I, before and after unilateral exposure to UVR-B. After day 3 and 7 post-exposure, ocular tissues were extracted for the detection of NKR-1 protein level by ELISA.Results: Pretreatment with Fosaprepitant decreases NKR-1 expression in exposed ocular tissues as well as in the unexposed lens epithelium compared to the saline group. Spantide I treatment showed a tendency of NKR-1 overexpression in ocular tissues.Conclusion: The clinically approved NKR-1 receptor antagonist Fosaprepitant decreases NKR-1 protein expression effectively not only in the exposed but also in the unexposed partner eye in a UVR-B irradiation mouse model. No effect was seen on the protein concentration of pro-inflammatory cytokines/chemokines in either eye.
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Catarata/metabolismo , Cristalino/efectos de la radiación , Morfolinas/farmacología , Antagonistas del Receptor de Neuroquinina-1/farmacología , Traumatismos Experimentales por Radiación/metabolismo , Receptores de Neuroquinina-1/metabolismo , Rayos Ultravioleta/efectos adversos , Animales , Humor Acuoso/efectos de los fármacos , Humor Acuoso/metabolismo , Catarata/etiología , Coroides/efectos de los fármacos , Coroides/metabolismo , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/metabolismo , Córnea/metabolismo , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Femenino , Inyecciones Intraperitoneales , Iris/efectos de los fármacos , Iris/metabolismo , Cristalino/efectos de los fármacos , Cristalino/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Traumatismos Experimentales por Radiación/etiología , Retina/efectos de los fármacos , Retina/metabolismo , Sustancia P/análogos & derivados , Sustancia P/farmacologíaRESUMEN
OBJECTIVE: To evaluate the long-term outcome and efficacy of intravitreal injection of gentamicin and dexamethasone sodium phosphate (IVGD) or triamcinolone in end-stage glaucoma patients and determine pre-procedure prognostic indicators of success and post-operative complications. PROCEDURE: Medical records were reviewed for 108 dogs (108 eyes) treated with intravitreal gentamicin with or without dexamethasone sodium phosphate or triamcinolone for glaucoma between 2013 and 2018 with 3 months of minimum follow-up. Signalment and clinical findings, including type of glaucoma, pre-procedure intraocular pressure (IOP), chronicity, procedure protocol, and outcome were recorded. Success was defined as an intraocular pressure of ≤25 mm Hg at the time of last re-examination or no ocular hypotensive medications at 3 months or longer post-injection. RESULTS: The overall success rate for pharmacologic ablation was 95%. The success rate for dogs receiving no ocular hypotensive medications was 86%. Seventy-six eyes (70.4%) had primary glaucoma, and 32 eyes (29.6%) had secondary glaucoma. Age at the time of injection had no effect on initial success but did in final success (P =-.03) for dogs requiring repeat injections. Cocker Spaniels required the most repeat 2nd and 3rd injections (3/12 dogs) and (2/4 dogs), respectively. No preoperative variable significantly affected the success rate. The most common complications were phthisis bulbi (59.2%), corneal edema (25.9%), and ulcerative keratitis (22.3%). Uncontrolled IOP resulted in enucleation in two dogs (1.8%). CONCLUSIONS: Pharmacologic ablation has a high overall success rate in lowering IOP to ≤25 mm Hg short-term in blind, glaucomatous canine eyes. Type of glaucoma, pre-procedure IOP, chronicity, and protocol did not affect success.
Asunto(s)
Técnicas de Ablación/veterinaria , Cuerpo Ciliar/efectos de los fármacos , Dexametasona/análogos & derivados , Enfermedades de los Perros/cirugía , Gentamicinas/uso terapéutico , Glaucoma/veterinaria , Animales , Cuerpo Ciliar/cirugía , Dexametasona/administración & dosificación , Dexametasona/efectos adversos , Dexametasona/uso terapéutico , Enfermedades de los Perros/tratamiento farmacológico , Perros , Femenino , Gentamicinas/administración & dosificación , Gentamicinas/efectos adversos , Glaucoma/tratamiento farmacológico , Glaucoma/cirugía , Inyecciones Intravítreas/veterinaria , Masculino , Estudios RetrospectivosRESUMEN
Purpose: Elevated IOP can cause the development of glaucoma. The circadian rhythm of IOP depends on the dynamics of the aqueous humor and is synchronized with the circadian rhythm pacemaker, that is, the suprachiasmatic nucleus. The suprachiasmatic nucleus resets peripheral clocks via sympathetic nerves or adrenal glucocorticoids. However, the detailed mechanisms underlying IOP rhythmicity remain unclear. The purpose of this study was to verify this regulatory pathway. Methods: Adrenalectomy and/or superior cervical ganglionectomy were performed in C57BL/6J mice. Their IOP rhythms were measured under light/dark cycle and constant dark conditions. Ocular administration of corticosterone or norepinephrine was also performed. Localization of adrenergic receptors, glucocorticoid receptors, and clock proteins Bmal1 and Per1 were analyzed using immunohistochemistry. Period2::luciferase rhythms in the cultured iris/ciliary bodies of adrenalectomized and/or superior cervical ganglionectomized mice were monitored to evaluate the effect of the procedures on the local clock. The IOP rhythm of retina and ciliary epithelium-specific Bmal1 knockout mice were measured to determine the significance of the local clock. Results: Adrenalectomy and superior cervical ganglionectomy disrupted IOP rhythms and the circadian clock in the iris/ciliary body cultures. Instillation of corticosterone and norepinephrine restored the IOP rhythm. ß2-Adrenergic receptors, glucocorticoid receptors, and clock proteins were strongly expressed within the nonpigmented epithelia of the ciliary body. However, tissue-specific Bmal1 knock-out mice maintained their IOP rhythm. Conclusions: These findings suggest direct driving of the IOP rhythm by the suprachiasmatic nucleus, via the dual corticosterone and norepinephrine pathway, but not the ciliary clock, which may be useful for chronotherapy of glaucoma.
Asunto(s)
Ritmo Circadiano/fisiología , Corticosterona/farmacología , Presión Intraocular/fisiología , Norepinefrina/farmacología , Sistema Nervioso Simpático/fisiología , Factores de Transcripción ARNTL/metabolismo , Administración Oftálmica , Adrenalectomía , Animales , Células Cultivadas , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/metabolismo , Ritmo Circadiano/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Ganglionectomía , Inmunohistoquímica , Iris/efectos de los fármacos , Iris/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas Circadianas Period/metabolismo , Fotoperiodo , Receptores Adrenérgicos beta 2/metabolismo , Receptores de Glucocorticoides/metabolismo , Ganglio Cervical Superior/cirugía , Tonometría OcularAsunto(s)
Cuerpo Ciliar/patología , Infecciones Bacterianas del Ojo/diagnóstico , Enfermedades del Iris/diagnóstico , Melanoma/diagnóstico , Tuberculoma/diagnóstico , Tuberculosis Ocular/diagnóstico , Enfermedades de la Úvea/diagnóstico , Neoplasias de la Úvea/diagnóstico , Acetilcolinesterasa/sangre , Adulto , Antígenos Bacterianos/genética , Antituberculosos/uso terapéutico , Humor Acuoso/metabolismo , Proteínas Bacterianas/genética , Sedimentación Sanguínea , Cuerpo Ciliar/efectos de los fármacos , Diagnóstico Diferencial , Infecciones Bacterianas del Ojo/tratamiento farmacológico , Expresión Génica/fisiología , Humanos , Enfermedades del Iris/tratamiento farmacológico , Masculino , Microscopía Acústica , Reacción en Cadena de la Polimerasa , Tuberculoma/tratamiento farmacológico , Tuberculosis Ocular/tratamiento farmacológico , Enfermedades de la Úvea/tratamiento farmacológicoRESUMEN
PURPOSE: We tested the hypothesis that changes in accommodation after instillation of Phenylephrine Hydrochloride (PHCl) observed in some studies could be caused by changes in optics. METHODS: We performed two experiments to test the effects of PHCl on static and on dynamic accommodation in 8 and 6 subjects, respectively. Objective wavefront measurements were recorded of the static accommodation response to a stimulus at different distances or dynamic accommodation response to a sinusoidally moving stimulus (between 1 and 3 D of accommodative demand at 0.2Hz). The responses were characterized using two methods: one that takes into account the mydriatic optical effects on the accommodation produced by higher-order aberrations of the eye and another that takes into account only power changes paraxially due to the action of the ciliary muscle and regardless of the pupil size. RESULTS: When mydriatic optical effects were taken into account, differences in responses before and after PHCl instillation were 0.51±0.53 D, and 0.12±0.15, for static and dynamic accommodation, respectively, and were statistically significant (p<0.039). When mydriatic optical effects were not taken into account, the differences in responses before and after PHCl instillation were -0.20±0.51 D, and -0.05±0.14, for static and dynamic accommodation, respectively, and were not statistically significant (p>0.313). CONCLUSIONS: The mydriatic effect of the PHCl causes optical changes in the eye that can reduce the objective and subjective measurement of accommodation.
Asunto(s)
Acomodación Ocular/efectos de los fármacos , Midriáticos/farmacología , Fenilefrina/farmacología , Adulto , Cuerpo Ciliar/efectos de los fármacos , Femenino , Humanos , MasculinoRESUMEN
The aim of this study was to investigate the effects of pilocarpine-induced ciliary body constant contraction for a long time period on the elastic modulus and collagen in corneal and scleral tissues in the early developmental stage. Twelve one-month-old New Zealand white rabbits were randomly monocularly treated with pilocarpine to cause ciliary body constant contraction. After 1- and 2-months, the sclera tissues and the eyeballs were obtained to assess the cornea and three regions of the sclera-anterior, equatorial and posterior. The corneal tissues and the three regions of the scleral tissues were cut into strips for elastic modulus measurement using an Instron 5544, the size distribution of collagen fibrils was examined using electron microscopy, the samples were homogenized, and the concentration of hydroxyproline was measured to determine the collagen content. For corneal tissues, long-term pilocarpine-induced ciliary body constant contraction did not affect the elastic modulus and collagen. For scleral tissues, long-term pilocarpine-induced ciliary body constant contraction affected the elastic modulus, diameter of collagen fibrils and collagen content in the equatorial and posterior sclera, however, the anterior sclera were unaffected. These results suggested that pilocarpine-induced ciliary body contraction could affect the scleral structure and lead to deformation of the sclera and eyeball, thereby affecting visual functions function in the process of ocular emmetropization.
Asunto(s)
Cuerpo Ciliar/metabolismo , Colágeno/metabolismo , Córnea/crecimiento & desarrollo , Módulo de Elasticidad , Pilocarpina/farmacología , Esclerótica/crecimiento & desarrollo , Animales , Cuerpo Ciliar/efectos de los fármacos , Colágeno/efectos de los fármacos , Colágeno/ultraestructura , Córnea/efectos de los fármacos , Córnea/metabolismo , Córnea/ultraestructura , Hidroxiprolina/metabolismo , Conejos , Esclerótica/efectos de los fármacos , Esclerótica/metabolismo , Esclerótica/ultraestructuraRESUMEN
Absence of the developing lens results in severe eye defects, including substantial reductions in eye size. How the lens controls eye expansion and the underlying signalling pathways are very poorly defined. We identified RDH10, a gene crucial for retinoic acid synthesis during embryogenesis, as a key factor downregulated in the peripheral retina (presumptive ciliary body region) of lens-removed embryonic chicken eyes prior to overt reductions in eye size. This is associated with a significant decrease in retinoic acid synthesis by lens-removed eyes. Restoring retinoic acid signalling in lens-removed eyes by implanting beads soaked in retinoic acid or retinal, but not vitamin A, rescued eye size. Conversely, blocking retinoic acid synthesis decreased eye size in lens-containing eyes. Production of collagen II and collagen IX, which are major vitreal proteins, is also regulated by the lens and retinoic acid signalling. These data mechanistically link the known roles of both the lens and retinoic acid in normal eye development, and support a model whereby retinoic acid production by the peripheral retina acts downstream of the lens to support vitreous production and eye expansion.
Asunto(s)
Cristalino/embriología , Cristalino/metabolismo , Transducción de Señal , Tretinoina/metabolismo , Aldehído Deshidrogenasa/metabolismo , Animales , Tipificación del Cuerpo/efectos de los fármacos , Embrión de Pollo , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/metabolismo , Colágeno/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Cristalino/anatomía & histología , Cristalino/efectos de los fármacos , Tamaño de los Órganos/efectos de los fármacos , Epitelio Pigmentado de la Retina/efectos de los fármacos , Epitelio Pigmentado de la Retina/metabolismo , Tenascina/metabolismo , Tretinoina/farmacología , Vitamina A/farmacologíaRESUMEN
The leaves of Perilla frutescens var. acuta (PFA) are commonly used as a traditional medicine in Korea, Japan, and China. We previously showed that PFA attenuates eye fatigue by improving visual accommodation through a clinical study. However, detailed mechanisms and chemical compounds have not been studied. In this study, we analyzed the active compounds in an aqueous extract of PFA involved in ciliary muscle relaxation in vitro and in vivo. NMR and MS analyses showed that the PFA extract contained mainly luteolin-7-O-diglucuronide and apigenin-7-O-diglucuronide. The composition after freeze-drying and spray-drying was similar. Freeze-dried PFA (50 µg/mL, 100 µg/mL, and 200 µg/mL) increased nitric oxide and cGMP levels in ciliary muscle cells isolated from the eyes of rats. [Ca2+]i decreased in a dose-dependent manner. Furthermore, Sprague-Dawley rats treated with freeze-dried PFA (200 mg/kg, orally) showed significantly increased cGMP levels compared with the control group and irradiated with white light. Our results suggest that PFA extract has the potential to reduce eye fatigue by relaxing ciliary muscles.
Asunto(s)
Cuerpo Ciliar/efectos de los fármacos , GMP Cíclico/metabolismo , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Óxido Nítrico/metabolismo , Perilla frutescens/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Animales , Calcio/metabolismo , Cromatografía Líquida de Alta Presión , RatasRESUMEN
PURPOSE: To evaluate the influence of prostaglandin (PG) analogues on the ciliary zonular fibers of the crystalline lens using scanning electron microscopy (SEM) of rabbit eyes, and to measure the matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) levels of the aqueous humor and crystalline lens treated with topical PG analogues Methods: Fifty eyes from 25 New Zealand white rabbits were divided into five groups of five rabbits each. In the control group, balanced salt solution was administered via the topical route once a day to the eyes. The benzalkonium chloride (BAC) group was treated with 0.02% BAC, the Latanoprost group with 0.005% latanoprost, the Travoprost group with 0.004% Travoprost, and the Bimatoprost group with 0.03% Bimatoprost for 10 months. We examined the ciliary zonular fibers using SEM. We also measured the MMP and TIMP levels of the aqueous humor and crystalline lens. RESULTS: SEM revealed some splitting of zonular fibers in eyes treated with topical PG analogues when compared with the control and BAC groups. The MMP-1 and TIMP-1 levels after treatment with the PG analogues did not differ significantly from the control and BAC groups (P > 0.05). There was no significant difference in MMP-1, MMP-3, TIMP-1, and MMP-1/TIMP-1 levels in the lens among all five groups. CONCLUSIONS: PG analogues may induce zonular change in rabbits microscopically. There was no association between zonular changes and the levels of certain types of MMP or TIMP in the aqueous humor or crystalline lens after topical treatment with PG analogues.
Asunto(s)
Bimatoprost/administración & dosificación , Cuerpo Ciliar/efectos de los fármacos , Glaucoma/tratamiento farmacológico , Presión Intraocular/efectos de los fármacos , Latanoprost/administración & dosificación , Cristalino/efectos de los fármacos , Prostaglandinas Sintéticas/administración & dosificación , Animales , Antihipertensivos/administración & dosificación , Humor Acuoso/metabolismo , Cuerpo Ciliar/ultraestructura , Modelos Animales de Enfermedad , Glaucoma/diagnóstico , Glaucoma/metabolismo , Presión Intraocular/fisiología , Cristalino/metabolismo , Cristalino/ultraestructura , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Microscopía Electrónica de Rastreo , Soluciones Oftálmicas , Conejos , Inhibidor Tisular de Metaloproteinasa-1/metabolismoRESUMEN
BACKGROUND Molecular hydrogen (H2) has been widely reported to have benefiicial effects in diverse animal models and human disease through reduction of oxidative stress and inflammation. The aim of this study was to investigate whether hydrogen gas could ameliorate endotoxin-induced uveitis (EIU) in rats. MATERIAL AND METHODS Male Sprague-Dawley rats were divided into a normal group, a model group, a nitrogen-oxygen (N-O) group, and a hydrogen-oxygen (H-O) group. EIU was induced in rats of the latter 3 groups by injection of lipopolysaccharide (LPS). After that, rats in the N-O group inhaled a gas mixture of 67% N2 and 33% O2, while those in the H-O group inhaled a gas mixture of 67% H2 and 33% O2. All rats were graded according to the signs of uveitis after electroretinography (ERG) examination. Protein concentration in the aqueous humor (AqH) was measured. Furthermore, hematoxylin-eosin staining and immunostaining of anti-ionized calcium-binding adapter molecule 1 (Iba1) in the iris and ciliary body (ICB) were carried out. RESULTS No statistically significant differences existed in the graded score of uveitis and the b-wave peak time in the Dark-adapted 3.0 ERG among the model, N-O, and H-O groups (P>0.05), while rats of the H-O group showed a lower concentration of AqH protein than that of the model or N-O group (P<0.05). The number of the infiltrating cells in the ICB of rats from the H-O group was not significantly different from that of the model or N-O group (P>0.05), while the activation of microglia cells in the H-O group was somewhat reduced (P<0.05). CONCLUSIONS Post-treatment hydrogen gas inhalation did not ameliorate the clinical signs, or reduce the infiltrating cells of EIU. However, it inhibited the elevation of protein in the AqH and reduced the microglia activation.
Asunto(s)
Hidrógeno/uso terapéutico , Uveítis/terapia , Animales , Humor Acuoso/efectos de los fármacos , Proteínas de Unión al Calcio/efectos de los fármacos , Cuerpo Ciliar/efectos de los fármacos , Modelos Animales de Enfermedad , Endotoxinas/efectos adversos , Hidrógeno/administración & dosificación , Hidrógeno/fisiología , Iris/efectos de los fármacos , Lipopolisacáridos/farmacología , Masculino , Proteínas de Microfilamentos/efectos de los fármacos , Microglía/efectos de los fármacos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ratas , Ratas Sprague-Dawley , Uveítis/inducido químicamenteRESUMEN
PURPOSE: To study the pharmacological profile of the serotonin (5-hydroxytryptamine [5-HT]) receptor subtype mediating contractions in bovine isolated ciliary muscles. METHODS: Ciliary muscle strips were isolated from bovine eyeballs and mounted in organ baths containing aerated (95% O2, 5% CO2) Krebs buffer solution maintained at 37°C. Each muscle strip was attached at 1 end to a Grass Force-displacement Transducer connected to a Polyview Computer System for recording changes in isometric tension. After an equilibration period, ciliary muscle strips were exposed to selective agonists and antagonists of 5-HT receptors. RESULTS: Both selective and nonselective agonists for 5-HT produced concentration-dependent contractions of isolated ciliary muscles with the following rank order of potency: BW723C86>α-methyl-5-HT>MK-212>>8-hydroxy-DPAT>quipazine>R-DOI>>5-HT>>tryptamine. The selective 5-HT2 receptor antagonists, M-100907 (5-HT2A), RS-127445 (5-HT2B), and RS-102221 (5-HT2C), produced noncompetitive inhibition of the contractile effects of selective agonists yielding antagonist potency (pKB) values of 251 ± 27.2 nM (n = 4), 52.5 ± 6.3 nM (n = 4), and 79.4 ± 9.5 nM (n = 4), respectively. CONCLUSION: On the basis of the profile of activity of selective agonists and antagonists, we conclude that the 5-HT2B and 5-HT2C receptor subtypes appear to be the predominant serotonin receptors that mediate the contractile action of this amine in bovine isolated ciliary muscles.
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Cuerpo Ciliar/efectos de los fármacos , Presión Intraocular/efectos de los fármacos , Contracción Muscular/efectos de los fármacos , Receptor de Serotonina 5-HT2B/metabolismo , Receptor de Serotonina 5-HT2C/metabolismo , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Animales , Bovinos , Cuerpo Ciliar/metabolismoAsunto(s)
Enfermedades de la Coroides/etiología , Cuerpo Ciliar/efectos de los fármacos , Queratoplastia Endotelial de la Lámina Limitante Posterior , Endotaponamiento/efectos adversos , Distrofia Endotelial de Fuchs/cirugía , Errores de Medicación , Hexafluoruro de Azufre/uso terapéutico , Enfermedades de la Úvea/etiología , Enfermedades de la Coroides/diagnóstico por imagen , Enfermedades de la Coroides/cirugía , Femenino , Humanos , Implantación de Lentes Intraoculares , Microscopía Acústica , Persona de Mediana Edad , Facoemulsificación , Tomografía de Coherencia Óptica , Enfermedades de la Úvea/diagnóstico por imagen , Enfermedades de la Úvea/cirugíaRESUMEN
Mineralocorticoid receptor (MR) contributes to retinal/choroidal homeostasis. Excess MR activation has been shown to be involved in pathogenesis of central serous chorioretinopathy (CSCR). Systemic administration of MR antagonist (MRA) reduces subretinal fluid and choroidal vasodilation, and improves the visual acuity in CSCR patients. To achieve long term beneficial effects in the eye while avoiding systemic side-effects, we propose the use of biodegradable spironolactone-loaded poly-lactic-co-glycolic acid (PLGA) microspheres (MSs). In this work we have evaluated the ocular tolerance of MSs containing spironolactone in rat' eyes. As previous step, we have also studied the tolerance of the commercial solution of canrenoate salt, active metabolite of spironolactone. PLGA MSs allowed in vitro sustained release of spironolactone for 30days. Rat eyes injected with high intravitreous concentration of PLGA MSs (10mg/mL) unloaded and loaded with spironolactone maintained intact retinal lamination at 1month. However enhanced glial fibrillary acidic protein immunostaining and activated microglia/macrophages witness retinal stress were observed. ERG also showed impaired photoreceptor function. Intravitreous PLGA MSs concentration of 2mg/mL unloaded and loaded with spironolactone resulted well tolerated. We observed reduced microglial/macrophage activation in rat retina compared to high concentration of MSs with normal retinal function according to ERG. Spironolactone released from low concentration of MSs was active in the rat retina. Low concentration of spironolactone-loaded PLGA MSs could be a safe therapeutic choice for chorioretinal disorders in which illicit MR activation could be pathogenic.
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Ácido Láctico/administración & dosificación , Antagonistas de Receptores de Mineralocorticoides/administración & dosificación , Ácido Poliglicólico/administración & dosificación , Espironolactona/administración & dosificación , Animales , Ácido Canrenoico/administración & dosificación , Cuerpo Ciliar/anatomía & histología , Cuerpo Ciliar/efectos de los fármacos , Liberación de Fármacos , Inyecciones Intravítreas , Ácido Láctico/química , Macrófagos/efectos de los fármacos , Masculino , Microglía/efectos de los fármacos , Microesferas , Antagonistas de Receptores de Mineralocorticoides/química , Antagonistas de Receptores de Mineralocorticoides/farmacocinética , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas Wistar , Retina/anatomía & histología , Retina/efectos de los fármacos , Retina/fisiología , Espironolactona/química , Espironolactona/farmacocinéticaRESUMEN
BACKGROUND: The literature is scant on the state of the ciliary body, its role in the development of rhegmatogenous retinal detachment (RRD) complicated by choroidal detachment (CD), and on ciliary body changes following the treatment aimed at resolving concomitant inflammation and choroidal attachment. This study assesses the anatomical position and thickness of the ciliary body and investigates the ciliary body changes after anti-inflammatory pre-vitrectomy treatment in RRD complicated by CD. METHODS: Forty-nine patients (49 eyes) with RRD complicated by CD underwent standard ophthalmological examination (including visual acuity assessment, biomicroscopy, ophthalmoscopy, and ocular tonometry) and ultrasound biomicroscopy of the ciliary body, choroid, and retina both before and following anti-inflammatory pre-vitrectomy treatment. RESULTS: At baseline, all subject eyes had ciliary body edema and detachment extending into the choroid. Ultrasonographic ciliary features included ciliary body edema and disorganization of the supraciliary layer of the pars plana, which was evident by the presence of multiple small oblique fibers. In all subject eyes, the treatment resulted in reattachment of the choroid and the ciliary body as well as a reduction in ciliary body edema (total mean ciliary thickness reduced from 0.83 (0.09) to 0.65 (0.09) mm, with a difference of 0.18 (0.07) mm, P < 0.001). CONCLUSIONS: Preoperative anti-inflammatory treatment in RRD complicated by CD results in restoration of the anatomical position of the ciliary body and a statistically significant reduction in ciliary body edema.
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Antiinflamatorios/uso terapéutico , Enfermedades de la Coroides/etiología , Coroides/diagnóstico por imagen , Cuerpo Ciliar/diagnóstico por imagen , Cuidados Preoperatorios/métodos , Desprendimiento de Retina/terapia , Vitrectomía , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades de la Coroides/diagnóstico , Cuerpo Ciliar/efectos de los fármacos , Femenino , Humanos , Masculino , Microscopía Acústica , Persona de Mediana Edad , Oftalmoscopía , Desprendimiento de Retina/complicaciones , Desprendimiento de Retina/diagnóstico , Estudios Retrospectivos , Resultado del Tratamiento , Agudeza Visual , Adulto JovenRESUMEN
We evaluated the safety of matrix metalloproteinase (MMP) inhibitor in experimental glaucoma filtration surgery in an animal model. Fifteen New Zealand white rabbits underwent an experimental trabeculectomy and were randomly allocated into 3 groups according to the adjuvant agent: no treatment group (n = 5), 0.02% mitomycin C (MMC) soaking group (n = 5), and MMP inhibitor (ilomastat) subconjunctival injection group (n = 5). Slit lamp examination with Seidel testing, pachymetry, and specular microscopy was performed preoperatively and postoperatively. The conjunctiva and ciliary body toxicity were evaluated with scores according to the pathologic grading systems. Electron microscopy was used to examine the structural changes in cornea, conjunctiva, and ciliary body. In the ilomastat-treated group, there was no statistically significant change in central corneal thickness preoperatively and at 28 days postoperatively (P = 0.655). There were also no significant changes in specular microscopy findings over the duration of the study in the ilomastat-treated group. The conjunctival toxicity score was 1 in the control group, 1.5 in the ilomastat-treated group, and 2 in the MMC-treated group. When assessing ciliary body toxicity scores, the ilomastat-treated group score was 0.5 and the MMC-treated group score was 1.5. Transmission electron microscopy did not show structural changes in the cornea and ciliary body whereas the structural changes were noticed in MMC group. A single subconjunctival injection of MMP inhibitor during the experimental trabeculectomy showed a less toxic affect in the rabbit cornea, conjunctiva, and ciliary body compared to MMC.
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Conjuntiva/efectos de los fármacos , Glaucoma/cirugía , Indoles/toxicidad , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Animales , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/fisiología , Conjuntiva/fisiología , Córnea/efectos de los fármacos , Córnea/fisiología , Ácidos Hidroxámicos , Microscopía Electrónica de Transmisión , Mitomicina/toxicidad , Conejos , TrabeculectomíaRESUMEN
Insulin has been shown to elicit changes of Na,K-ATPase activity in various tissues. Na,K-ATPase in the nonpigmented ciliary epithelium (NPE) plays a role in aqueous humor secretion and changes of Na,K-ATPase activity impact the driving force. Because we detect a change of NPE Na,K-ATPase activity in response to insulin, studies were carried out to examine the response mechanism. Ouabain-sensitive rubidium (Rb) uptake by cultured NPE cells, measured as a functional index of Na,K-ATPase-mediated inward potassium transport, was found to increase in cells exposed for 5 min to insulin. The maximally effective concentration was 100 nM. An intrinsic increase of Na,K-ATPase activity evident as a >2-fold increase in the rate of ouabain-sensitive ATP hydrolysis in homogenates obtained from cells exposed to 100 nM insulin for 5 min was also observed. Insulin-treated cells exhibited Akt, Src family kinase (SFK), ERK1/2, and p38 activation, all of which were prevented by a pI3 kinase inhibitor LY294002. The Rb uptake and Na,K-ATPase activity response to insulin both were abolished by PP2, an SFK inhibitor which also prevented p38 and ERK1/2 but not Akt activation. The Akt inhibitor MK-2206 did not change the Na,K-ATPase response to insulin. The findings suggest insulin activates pI3K-dependent Akt and SFK signaling pathways that are separate. ERK1/2 and p38 activation is secondary to and dependent on SFK activation. The increase of Na,K-ATPase activity is dependent on activation of the SFK pathway. The findings are consistent with previous studies that indicate a link between Na,K-ATPase activity and SFK signaling. J. Cell. Physiol. 232: 1489-1500, 2017. © 2016 Wiley Periodicals, Inc.
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Cuerpo Ciliar/metabolismo , Epitelio/metabolismo , Insulina/metabolismo , Pigmentación , Transducción de Señal , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Familia-src Quinasas/metabolismo , Animales , Butadienos/farmacología , Cromonas/farmacología , Cuerpo Ciliar/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Epitelio/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Compuestos Heterocíclicos con 3 Anillos/farmacología , Imidazoles/farmacología , Insulina/farmacología , Modelos Biológicos , Morfolinas/farmacología , Nitrilos/farmacología , Ouabaína/farmacología , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Fosforilación/efectos de los fármacos , Pigmentación/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Piridinas/farmacología , Pirimidinas/farmacología , Rubidio/metabolismo , Transducción de Señal/efectos de los fármacos , Sus scrofa , Factores de Tiempo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
The therapeutic efficacy of rapamycin conjugated monomethoxy poly(ethylene glycol)-poly(ε-caprolactone) (MPEG-PCL) micelles (rapamycin micelles) was evaluated in a rat experimental autoimmune uveitis (EAU) model. Rapamycin micelles exhibited spherical morphology and had a mean particle size of 40nm and a zeta-potential of -0.89mv. The water solubility of rapamycin improved by more than 1000-fold in a micellar formulation. Intravitreal injection of MPEG-PCL micelles did not result in vitreous hemorrhage or retinal detachment. Fluorescence microscopy demonstrated that labeled micelles localized to the retinal pigment epithelium for at least 14 days following injection and the drug concentration of rapamycin micelles in the retinal tissue was significantly higher than unconjugated rapamycin over this period. At the optimal concentration of rapamycin micelles (9µg/eye), clinical signs of EAU were abolished via the downregulation of the Th1 and Th17 response. There were no significant difference in T cell proliferation and delayed-type hypersensitivity between the treatment and control groups, suggesting that the therapeutic effect of rapamycin manifested locally in the eye and not systemically. These results indicate that intravitreal injection of rapamycin micelles is a promising therapy for controlling sterile intraocular inflammation.
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Enfermedades Autoinmunes/tratamiento farmacológico , Inmunosupresores/administración & dosificación , Micelas , Sirolimus/administración & dosificación , Uveítis/tratamiento farmacológico , Animales , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/metabolismo , Enfermedades Autoinmunes/patología , Proliferación Celular/efectos de los fármacos , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/patología , Citocinas/inmunología , Modelos Animales de Enfermedad , Inmunosupresores/farmacocinética , Inmunosupresores/farmacología , Inmunosupresores/uso terapéutico , Inyecciones Intravítreas , Poliésteres/química , Polietilenglicoles/química , Ratas Sprague-Dawley , Retina/efectos de los fármacos , Retina/metabolismo , Retina/patología , Sirolimus/farmacocinética , Sirolimus/farmacología , Sirolimus/uso terapéutico , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Uveítis/inmunología , Uveítis/metabolismo , Uveítis/patología , Cuerpo Vítreo/metabolismoRESUMEN
INTRODUCTION: Experimental animal models of acute uveitis, an inflammatory eye disease, can be established via endotoxin-induced inflammation. Propolis, a natural substance collected by honeybees from buds and tree exudates, has antioxidant, antibacterial, antiviral, and anti-inflammatory effects. We investigated the effects of propolis, obtained from the Sakarya province of Turkey, on endotoxin-induced uveitis using immunohistochemical, ultrastructural, and biochemical approaches. MATERIAL AND METHODS: Male Wistar albino rats (n = 6/group) received intraperitoneal (ip) lipopolysaccharide (LPS) endotoxin (150 µg/kg) followed by aqueous extract of propolis (50 mg/kg ip) or vehicle; two additional groups received either saline (control) or propolis only. After 24 h, aqueous humor (AH) was collected from both eyes of each animal for analysis of tumor necrosis factor-α (TNF-α) and hypoxia-inducible factor-1α (HIF-1α). Right eyeballs were paraffin-embedded for immunohistochemical staining of nuclear factor κB (NF-κB)/p65 and left eyeballs were araldite-embedded for ultrastructural analysis. RESULTS: Treatment of LPS-induced uveitis with propolis significantly reduced ciliary body NF-κB/p65 immunoreactivity and AH levels of HIF-1α and TNF-α. Ultrastructural analysis showed fewer vacuoles and reduced mitochondrial degeneration in the retinal pigment epithelium, as compared to the uveitis group. The intercellular spaces of the inner nuclear layer and outer limiting membrane were comparable with those of the control group; no polymorphonuclear cells or stasis was observed in intravascular or extravascular spaces. CONCLUSIONS: This is the first report demonstrating an anti-inflammatory effect of Turkish propolis in a rat model of LPS-induced acute uveitis, suggesting a therapeutic potential of propolis for the treatment of inflammatory ophthalmic diseases.