RESUMEN
PURPOSE: Epidemic keratoconjunctivitis (EKC) is a contagious acute conjunctivitis associated with community-acquired infection. Human adenovirus type 8 (HAdV-8) is one of the major serotypes isolated from patients with EKC. DNA restriction enzyme analyses were performed to investigate the genetic characteristics of the isolates and their chronological pattern. METHODS: Viral samples were taken from 11 strains isolated from sporadic cases of EKC and identified as HAdV-8 by the neutralization method with type-specific antiserum against HAdV-8 between 1986 and 2003 in Japan. DNA restriction enzyme analysis included six restriction enzymes: BamHI, HindIII, PstI, SacI, SalI, and SmaI. RESULTS: The restriction patterns revealed that the genome types were HAdV-8A and HAdV-8B in 1986, HAdV-8K in 1991, and HAdV-8E in 1996. HAdV-8K was a new genome type revealed with the enzyme SacI. Two strains isolated in 2003 exhibited identical restriction patterns as HAdV-54, which was described in 2008 and collected from Japanese patients in 2000. CONCLUSIONS: Genetic changes might occur chronologically in HAdV-8. HAdV-8 displays considerable variability. The investigations of these variants might be helpful for defining the evolutionary tendency and to predict future outbreaks of HAdV infection.
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Adenovirus Humanos/genética , Epidemias , Variación Genética , Genoma Viral/genética , Queratoconjuntivitis/epidemiología , Queratoconjuntivitis/virología , Adenovirus Humanos/clasificación , Adenovirus Humanos/aislamiento & purificación , Humanos , Japón/epidemiología , Queratoconjuntivitis/sangre , Filogenia , Mapeo Restrictivo , SerotipificaciónAsunto(s)
Linfoma de Células B/fisiopatología , Regresión Neoplásica Espontánea , Neoplasias de la Retina/fisiopatología , Anciano de 80 o más Años , Antígenos CD20/metabolismo , Linfocitos T CD8-positivos/patología , Enucleación del Ojo , Reordenamiento Génico , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Interleucina-10/metabolismo , Linfoma de Células B/genética , Linfoma de Células B/metabolismo , Masculino , Reacción en Cadena de la Polimerasa , Neoplasias de la Retina/genética , Neoplasias de la Retina/metabolismo , Agudeza Visual/fisiología , Vitrectomía , Cuerpo Vítreo/metabolismoRESUMEN
AIMS: Hydrogen peroxide (H(2)O(2)) is the major oxidant involved in cataract formation. Lens epithelial cells have been suggested to be the first site of oxidative damage. The authors investigated the relationship between H(2)O(2)-induced cytotoxicity and activation of nuclear factor kappa B (NF-kappaB) in human lens epithelial (HLE) cells. METHODS: HLE B-3 cells were stimulated by various concentrations of H(2)O(2) in the presence or absence of pyrrolidine dithiocarbamate (PDTC), a potent inhibitor of NF-kappaB. H(2)O(2)-induced cytotoxicity was measured by lactate dehydrogenase cytotoxicity assay. Translocation of NF-kappaB was examined by Western blot and immunocytochemistry using anti-p65 antibody. RESULTS: H(2)O(2)-induced cytotoxicity increased in a concentration-dependent manner. PDTC treatment significantly suppressed the cytotoxicity induced by H(2)O(2). After stimulated with H(2)O(2), NF-kappaB was found translocated from cytoplasm into the nuclei. PDTC treatment also inhibited the translocation of NF-kappaB. CONCLUSIONS: NF-kappaB signal pathway may be important in the development of H(2)O(2)-induced damage in HLE cells that is involved in cataractogenesis.
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Peróxido de Hidrógeno/antagonistas & inhibidores , Cristalino/efectos de los fármacos , FN-kappa B/fisiología , Western Blotting , Muerte Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Peróxido de Hidrógeno/farmacología , Cristalino/citología , Cristalino/metabolismo , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Pirrolidinas/farmacología , Tiocarbamatos/farmacología , Translocación Genética/efectos de los fármacosRESUMEN
Erythropoietin (Epo) induces physiological activities such as cell proliferation, migration, and angiogenesis in Epo receptor (EpoR)-expressing vascular endothelial and tumor cells. Recently, it has been demonstrated that growth factor-independent proliferation is frequently observed during the cell transformation process. Pterygium is a fibrovascular proliferating tissue that includes transformed cells. The aim of this study was to examine the localization of Epo and EpoR proteins in human pterygial tissues. Eleven samples including nine pterygia and two normal bulbar conjunctivas, which were surgically excised, were studied. Formalin-fixed, paraffin-embedded tissue sections were constructed and then were examined by immunohistochemistry with anti-Epo and EpoR antibodies. Cytoplasmic immunoreactivity for EpoR was heterogeneously detected in basal and suprabasal cells of the pterygium epithelium. In the pterygium stroma, a variety of endothelial cells forming vascular cavities showed cytolasmic immunoreactivity for EpoR. In normal conjunctival epithelium, a few basal cells showed a weak homogeneous immunoreactivity for EpoR in the cytoplasm. The number of EpoR-expressing epithelial cells was much higher in the pterygium compared to the normal conjunctiva. EpoR expression was marginally detected in stromal microvessels of the normal conjunctiva. Immunoreactivity for Epo was not noted in pterygium epithelium and stroma, and in normal conjunctiva. These results suggest that the Epo-independent EpoR-signaling pathway plays a potential role in cell proliferation and angiogenesis in human pterygium.
Asunto(s)
Pterigion/metabolismo , Receptores de Eritropoyetina/metabolismo , Conjuntiva/metabolismo , Conjuntiva/patología , Epitelio/metabolismo , Epitelio/patología , Humanos , Pterigion/patología , Receptores de Eritropoyetina/inmunologíaRESUMEN
PURPOSE: Lutein deposits in the macula and lens of human eyes with high concentration and is well known as an eye-protective nutrient for its beneficial effects on eye disease such as age-related macular degeneration and cataract. The purpose of the present study was to investigate the effects of lutein on endotoxin-induced uveitis (EIU) in rats. METHODS: EIU was induced in male Lewis rats by subcutaneous injection of 200 microg lipopolysaccharide. Lutein or dexamethasone was administered intravenously at 30 minutes before, at the same time as, and at 30 minutes after LPS treatment. The aqueous humor was collected at 24 hours after LPS injection, the number of infiltrating cells, the protein concentration, and the levels of nitric oxide (NO), tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, prostaglandin (PG)-E2, monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein (MIP)-2 in the aqueous humor were determined. Immunohistochemical staining with a monoclonal antibody against activated nuclear factor (NF)-kappaB was performed to evaluate the effect of lutein on NF-kappaB activation in the iris-ciliary body (ICB) of rats. A mouse macrophage cell line (RAW264.7 cells) was stimulated with LPS in the presence or absence of lutein. Expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and degradation of inhibitor kappaB (IkappaB) were analyzed by Western blot analysis. RESULTS: Lutein suppressed the development of EIU in a dose-dependent fashion. The anti-inflammatory effect of 100 mg/kg lutein was as strong as that of 1 mg/kg dexamethasone. Treatment with lutein reduced the concentrations of NO, TNF-alpha, IL-6, PGE2, MCP-1, and MIP-2 in aqueous humor. Lutein also suppressed the activation of NF-kappaB in the ICB as well as iNOS and COX-2 expression and IkappaB degradation in RAW cells. CONCLUSIONS: These findings indicate that lutein has anti-inflammatory effects on EIU by inhibiting the NF-kappaB dependent signaling pathway and the subsequent production of proinflammatory mediators.
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Antiinflamatorios no Esteroideos/farmacología , Luteína/farmacología , Uveítis Anterior/prevención & control , Animales , Humor Acuoso/metabolismo , Western Blotting , Línea Celular , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/metabolismo , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Proteínas I-kappa B/metabolismo , Iris/efectos de los fármacos , Iris/metabolismo , Lipopolisacáridos , Macrófagos/metabolismo , Masculino , Ratones , Microscopía Confocal , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ratas , Ratas Endogámicas Lew , Salmonella typhimurium , Uveítis Anterior/metabolismoRESUMEN
PURPOSE: To investigate the serotypes of adenovirus causing conjunctivitis in Hanoi, Vietnam. DESIGN: Clinical diagnosis of adenoviral conjunctivitis and laboratory-based experimental study. METHODS: We collected 21 conjunctival swabs from 21 different patients with a clinical presentation compatible with adenoviral conjunctivitis, in Hanoi, Vietnam. Immunochromatography and real-time polymerase chain reaction (PCR) were performed to detect human adenovirus (HAdV). The sequence of PCR products was analyzed to determine the serotype of HAdV. RESULTS: Of 21 samples, HAdV DNA was detected in 14 samples (66.7%) by real-time PCR. The serotype analysis showed HAdV-8 in 11 samples (78.6%), HAdV-3 in two samples (14.3%), and HAdV-37 in one sample (7.1%). Of 11 HAdV-8 samples, one sample (9.1%) was prototype, and the other 10 samples (90.9%) had identical nucleotide sequence and were identified as a variant of HAdV-8. CONCLUSIONS: HAdV-8 was found to be the predominant serotype in Hanoi, Vietnam. Most of the HAdV-8 samples were a variant of HAdV-8.
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Infecciones por Adenovirus Humanos/epidemiología , Adenovirus Humanos/genética , Conjuntivitis Viral/epidemiología , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/clasificación , Cromatografía , Conjuntiva/virología , Conjuntivitis Viral/virología , ADN Viral/genética , Humanos , Técnicas Inmunológicas , Epidemiología Molecular , Reacción en Cadena de la Polimerasa , Vietnam/epidemiologíaRESUMEN
Opacification of the posterior capsule depends on replication of the residual lens epithelial cells lining the capsule. However, the mechanisms in the regulation of lens cell proliferation have not been determined. The purpose of this study is to examine the expression of p27(KIP1), a cyclin-dependent kinase inhibitor, and its phosphorylation, and cyclin D1 in lens epithelial cells after extraction of fiber cells. C57Bl6 mice (12 weeks old) were anesthetized, and the lens fiber cells were surgically extracted. Eyeballs were collected and fixed at 15 min and 24 h after extraction with and without injection of a specific phosphorylated extracellular signal-regulated kinase (pERK) 1/2 inhibitor (PD98059) to the anterior chamber. Collected tissues were analyzed using immunohistochemistry with anti-p27(KIP1), anti-phosphorylated p27(KIP1) on serine 10 (s10-phospho-p27) and cyclin D1 antibodies. Human lens epithelial cells were cultured, and then were treated with and without 40 ng/ml human recombinant basic fibroblast growth factor (bFGF), which was analyzed by Western blot analysis. In the untreated lens, p27(KIP1) was not phosphorylated in the lens epithelial cells, although p27(KIP1)-positive nuclei were detected in the lens cells of the equatorial region. Immunoreactivity for cyclin D1 was hardly detected in the lens. Nuclear immunoreactivity for p27(KIP1) and s10-phospho-p27 was observed in several lens cells of the equatorial region 15 min after extraction of fiber cells. Western blotting demonstrated that the p27(KIP1) phosphorylation form was upregulated 15 min after bFGF treatment in cultured lens epithelial cells. Many cyclin D1-positive nuclei were noted 24 h after the surgery. p27(KIP1) phosphorylation and cyclin D1 induction were inhibited by PD98059. s10-phospho-p27 and p27(KIP1) immunoreactivity was undetected in the lens cells 24 h after the extraction of fiber cells. It is possible that the phosphorylation of p27(KIP1), and cyclin D1 expression are regulated by the ERK pathway in lens cells after the extraction of fiber cells.
Asunto(s)
Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Células Epiteliales/metabolismo , Cristalino/citología , Cristalino/metabolismo , Cristalino/cirugía , Animales , Western Blotting , Técnicas de Cultivo de Célula , Línea Celular Transformada , Transformación Celular Viral , Células Cultivadas , Ciclina D1/metabolismo , Inhibidores Enzimáticos/farmacología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Factor 2 de Crecimiento de Fibroblastos/farmacología , Flavonoides/farmacología , Humanos , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Fosforilación , Regulación hacia ArribaRESUMEN
PURPOSE: Aronia crude extract (ACE) with high levels of polyphenol compounds has been reported to have antioxidative effects in vitro and in vivo. In this study, attention was focused on the antioxidant effect of ACE. The purpose of the present study was to investigate the effect of ACE on endotoxin-induced uveitis (EIU) in rats. In addition, the endotoxin-induced expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins was investigated in a mouse macrophage cell line (RAW 264.7) treated with ACE in vitro, to clarify the anti-inflammatory effect. METHODS: EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, 1, 10, or 100 mg ACE or 10 mg prednisolone was injected intravenously. After 24 hours, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration, nitric oxide (NO), prostaglandin (PG)-E2, and TNF-alpha levels in the aqueous humor were determined. RAW 264.7 cells treated with various concentrations of ACE were incubated with 10 mug/mL LPS for 24 hours. Levels of NO, PGE2, and TNF-alpha were determined by an enzyme-linked immunosorbent assay. The expression of iNOS and COX-2 proteins was analyzed by Western blot analysis. RESULTS: The number of inflammatory cells, the protein concentrations, and the levels of NO, PGE2, and TNF-alpha in the aqueous humor in the groups treated with ACE were significantly decreased in a dose-dependent manner. In addition, the anti-inflammatory effect of 100 mg ACE was as strong as that of 10 mg prednisolone. The anti-inflammatory action of ACE was stronger than that of either quercetin or anthocyanin administered alone. ACE also suppressed LPS-induced iNOS and COX-2 protein expressions in RAW 264.7 cells in vitro in a dose-dependent manner. CONCLUSIONS: The results suggest that ACE has a dose-dependent anti-ocular inflammatory effect that is due to the direct blocking of the expression of the iNOS and COX-2 enzymes and leads to the suppression of the production of NO, PGE2, and TNF-alpha.
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Antiinflamatorios/uso terapéutico , Photinia/química , Fitoterapia , Extractos Vegetales/uso terapéutico , Uveítis Anterior/tratamiento farmacológico , Animales , Humor Acuoso/citología , Humor Acuoso/metabolismo , Western Blotting , Línea Celular , Ciclooxigenasa 2 , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática , Frutas , Inyecciones Intravenosas , Isoenzimas/metabolismo , Lipopolisacáridos/toxicidad , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Prednisolona/uso terapéutico , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas Endogámicas Lew , Salmonella typhimurium , Factor de Necrosis Tumoral alfa/metabolismo , Uveítis Anterior/inducido químicamente , Uveítis Anterior/enzimología , Uveítis Anterior/patologíaRESUMEN
The aim of this study was to investigate the efficacy of naringin and naringenin on endotoxin- induced uveitis (EIU) in rats. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). The rats were injected intravenously with 0.4, 4, or 40 microg/kg naringin or naringenin. Each compound was administered three times, simultaneously, 30 min before and after the actual LPS injection. The aqueous humor was collected 24 h later from both eyes, and the number of cells infiltrating into the aqueous humor and the aqueous humor protein concentration were measured. The levels of prostaglandin E2 (PGE2) and nitric oxide (NO) were determined. Naringin and naringenin suppressed the development of EIU in a dose-dependent fashion. Both treatments with naringin and naringenin produced reductions in PGE2 and NO concentrations in the aqueous humor. In particular, 40 microM/kg of naringin and naringenin suppressed increases in cell count owing to LPS treatment by 31% and 38%, respectively. The possible mechanism for the antiocular inflammatory effect may be the suppression of PGE2 and NO by naringin and naringenin.
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Humor Acuoso/efectos de los fármacos , Flavanonas/uso terapéutico , Lipopolisacáridos/toxicidad , Uveítis Anterior/prevención & control , Animales , Humor Acuoso/citología , Humor Acuoso/metabolismo , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Quimioterapia Combinada , Flavanonas/administración & dosificación , Lipopolisacáridos/aislamiento & purificación , Masculino , Óxido Nítrico/metabolismo , Ratas , Ratas Endogámicas Lew , Salmonella typhimurium/metabolismo , Uveítis Anterior/inducido químicamente , Uveítis Anterior/patologíaRESUMEN
BACKGROUND: We report a case of bacterial endophthalmitis caused by an intraocular cilium in a patient without any history of trauma or ocular surgery. FINDINGS: A 32-year-old Caucasian male showed symptoms of orbital myositis and scleritis, with no intraocular inflammation in the right eye. The patient had been treated with infliximab for Crohn's disease. Three weeks after initiation of oral prednisolone therapy, he developed bacterial endophthalmitis. During pars plana vitrectomy, a cilium in the massive vitreous opacity was found. A focal scleral necrosis was detected just outside where the cilium was intraoperatively observed. Vitreous culture showed the presence of Staphylococcus aureus. CONCLUSIONS: The intraocular cilium seemed to be the aetiology of the endophthalmitis in this case, which suggests that anti-tumour necrosis factor-α therapy may play a role in the migration of cilia into the globe and the occurrence of endophthalmitis.
RESUMEN
Human adenoviruses (HAdVs) have been implicated in a wide range of diseases affecting primarily the respiratory, ocular and gastrointestinal systems. A rapid and efficient method for the detection of HAdV hexon antigen is described using carbon nanotube (CNT) sensors. Anti-HAdV antibody was immobilised on the reverse surface of a CNT sensor. As a control, non-specific mouse IgG was immobilised on another CNT sensor. I-V(gate) curves were measured after incubation of various concentrations of recombinant HAdVs hexon antigen with anti-HAdVs antibody-immobilised or non-specific mouse IgG-immobilised sensors. The curves showed a positive shift that was dependent on the hexon antigen concentration in the anti-HAdV antibody-immobilised sensor, whereas no such shift was observed in the non-specific mouse IgG-immobilised sensor. The sensitivity of the CNT sensor method was greater than that of enzyme-linked immunosorbent assay. Hence, this method offers a new tool for HAdV detection by analysing antigen-antibody interactions.
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Adenovirus Humanos/aislamiento & purificación , Antígenos Virales/análisis , Proteínas de la Cápside/análisis , Nanotubos de Carbono , Virología/métodos , Animales , Técnicas Biosensibles/métodos , Humanos , Ratones , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Angiotensin II type 1 (AT1) receptor-antagonists are widely used for treatment of hypertension. Recent studies have demonstrated a protective effect of renin angiotensin system (RAS) antagonism against immune-mediated inflammatory diseases such as myocarditis, chronic allograft rejection, antiglomerular basement membrane nephritis, colitis, and arthritis. However, only a few reports have demonstrated the effect of RAS in ocular inflammatory conditions. The purpose of this study was to investigate the anti-inflammatory effect of a selective AT1 receptor antagonist, losartan, on endotoxin-induced uveitis (EIU) and compare the effect on experimental autoimmune uveoretinitis (EAU). METHODS: To induce EIU, 7-week-old Lewis rats were injected subcutaneously with 200 microg lipopolysaccharide (LPS). Losartan was administered intravenously at the same time. The aqueous humor was collected from eyes 24 h after LPS injection. The number of infiltrating cells, protein concentration, and levels of tumor necrosis factor (TNF)-alpha and monocyte chemoattractant protein-1 (MCP-1) in the aqueous humor were determined. The collected eyes were immunohistochemically stained with monoclonal antibody for activated nuclear factor (NF)-kappaB. To induce EAU, C57BL/6 mice (6-8 weeks old) were immunized with human interphotoreceptor retinoid binding protein (hIRBP)-derived peptide emulsified in complete Freund's adjuvant (CFA) and concomitantly injected with purified Bordetella pertussis toxin (PTX). Clinical severity of EAU and T cell proliferative response were analyzed. RESULTS: Losartan significantly suppressed the development of EIU. Numbers of aqueous cells of control EIU rats, those from EIU rats treated with 1 or 10 mg/kg of losartan were 75.3+/-45.6 x 10(5), 27.9+/-8.1 x 10(5), or 41.3+/-30.9 x 10(5) cells/ml respectively (p<0.01 vs control). Aqueous protein, TNF-alpha, and MCP-1 levels were also significantly decreased in a manner dependent on the amount of losartan administered (p<0.01). Treatment of EIU rats with losartan suppressed activation of NF-kappaB at the iris ciliary body. Thus, the suppressive effect of losartan on ocular inflammation in EIU appeared to result from down-regulation of NF-kappaB activation and reduction of inflammatory cytokine production. On the other hand, in the EAU model, neither the clinical score nor the antigen-specific T cell proliferative response was significantly influenced by the treatment with losartan. CONCLUSIONS: The present findings indicate that RAS may be involved in the acute inflammation of the eye, but not in T cell-dependent ocular autoimmunity. Antagonism of the RAS may be a potential prophylactic strategy for treatment of the human acute ocular inflammation.
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Bloqueadores del Receptor Tipo 1 de Angiotensina II/uso terapéutico , Enfermedades Autoinmunes/tratamiento farmacológico , Losartán/uso terapéutico , Retinitis/tratamiento farmacológico , Uveítis/tratamiento farmacológico , Enfermedad Aguda , Animales , Humor Acuoso/metabolismo , Enfermedades Autoinmunes/metabolismo , Enfermedades Autoinmunes/patología , Quimiocina CCL2/metabolismo , Cuerpo Ciliar/metabolismo , Cuerpo Ciliar/patología , Modelos Animales de Enfermedad , Femenino , Lipopolisacáridos , Activación de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratas , Ratas Endogámicas Lew , Sistema Renina-Angiotensina/fisiología , Retinitis/metabolismo , Retinitis/patología , Salmonella typhimurium , Linfocitos T/inmunología , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Uveítis/metabolismo , Uveítis/patologíaRESUMEN
Captopril is an inhibitor of angiotensin-converting enzyme (ACE) that is largely used in the treatment of cardiovascular diseases. Several previous studies have demonstrated that captopril exhibits a wide variety of biological activities, including an anti-inflammatory action, on which we focused our attention. The aim of the present study was to investigate the efficacy of captopril on endotoxin induced uveitis (EIU) in rats. We investigated its effect upon cellular infiltration and protein leakage, as well as on the concentration of tumor necrosis factor-alpha (TNF-alpha), nitric oxide (NO), prostaglandin E2 (PGE2), monocyte chemoattractant protein-1 (MCP-1) in the anterior chamber. In addition, we checked its effect on activation of nuclear factor kappa B (NF-kappaB) in iris and ciliary body (ICB) cells in vivo. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). One hour after the LPS inoculation, either 1mg/kg, 10mg/kg or 100mg/kg captopril were injected intravenously. 24h later, the aqueous humor was collected from both eyes, and the number of infiltrating cells and protein concentration in the aqueous humor were determined. Levels of TNF-alpha, PGE2, NO and MCP-1 were determined by enzyme-linked immunosorbent assay. On some eyes, after enucleation, immunohistochemical staining with a monoclonal antibody against activated NF-kappaB was performed. Captopril treatment significantly decreased the inflammatory cells infiltration, the level of protein, concentrations of TNF-alpha, PGE2, NO and MCP-1 in the aqueous humor. The number of activated NF-kappaB-positive cells was lower in ICB of the rats treated with captopril 3h after the LPS injection. The present results indicate that captopril suppresses the inflammation in EIU by inhibiting the NF-kappaB-dependent pathway and the subsequent production of pro-inflammatory mediators.
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Antiinflamatorios/uso terapéutico , Captopril/uso terapéutico , Úvea/inmunología , Uveítis/tratamiento farmacológico , Animales , Humor Acuoso/química , Quimiocina CCL2/análisis , Depresión Química , Dinoprostona/análisis , Inmunohistoquímica/métodos , Lipopolisacáridos , Masculino , Modelos Animales , FN-kappa B/análisis , Nitritos/análisis , Ratas , Ratas Endogámicas Lew , Úvea/efectos de los fármacos , Úvea/microbiología , Uveítis/inmunología , Uveítis/microbiologíaRESUMEN
The aim of the present study was to investigate the effects of blue honeysuckle extract (BHE), which contains high level of phenolic compounds, on endotoxin-induced uveitis (EIU). Male Lewis rats were randomly divided into 5 groups with 14 rats in each (eight rats for collection of aqueous humor, six rats for histologic examination). EIU was induced by a footpad injection of lipopolysaccharide (LPS). 1, 10, or 100 mg of BHE was injected intravenously immediately after LPS injection. The aqueous humor was collected at 24 h after LPS injection, the number of infiltrating cells, protein concentration, nitric oxide (NO), tumor necrosis factor (TNF)-alpha, and prostaglandin (PG)-E2 levels in the aqueous humor were determined. Some eyes were enucleated for histologic examination and immunohistochemical analysis. Immunohistochemical staining with a monoclonal antibody against activated nuclear factor (NF)-kappaB was performed to evaluate the effect of BHE on NF-kappaB activation. To further clarify the anti-inflammatory effect, RAW264.7 cells (a mouse macrophage cell line) were stimulated with LPS in the presence or absence of BHE and its major phenolics, cyanidin 3-glucoside (C3G), cyanidin 3-rutinoside (C3R), chlorogenic acid (CA). Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. BHE treatment significantly reduced the inflammatory cell infiltration, the protein concentration, the levels of NO, TNF-alpha and PGE2 in the aqueous humor and improved histologic status of the ocular tissue. The number of activated NF-kappaB-positive cells was lower in the iris-ciliary body treated with BHE at 3 h after LPS injection. BHE significantly suppressed the production of NO, PGE2 and TNF-alpha in the culture medium as well as the expression of iNOS and COX-2 by LPS-stimulated RAW264.7 cells in a dose-dependent fashion. C3G, C3R and CA showed no or weak inhibitory effects on the level of inflammatory mediators and the expression of iNOS and COX-2. These results suggest that BHE attenuates the degree of inflammation in eyes with EIU by inhibiting the NF-kappaB dependent signaling pathway and the subsequent production of proinflammatory mediators.
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Lonicera , Fitoterapia/métodos , Uveítis/tratamiento farmacológico , Animales , Humor Acuoso/metabolismo , Western Blotting , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ciclooxigenasa 2/metabolismo , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Proteínas del Ojo/metabolismo , Lipopolisacáridos , Masculino , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Ratas , Ratas Endogámicas Lew , Factor de Necrosis Tumoral alfa/metabolismo , Uveítis/inducido químicamente , Uveítis/metabolismo , Uveítis/patologíaRESUMEN
We investigated the effects of astaxanthin (AST), a carotenoid, on endotoxin-induced uveitis (EIU), and over the course of the disease measured the expression of inflammatory cytokines and chemokines in the presence or absence of AST. EIU was induced in male Lewis rats by footpad injection of lipopolysaccharide (LPS). The animals were randomly divided to 12 groups with eight animals in each. Immediately after the inoculation, AST (1, 10, or 100 mg kg(-1)) was injected intravenously. Aqueous humour was collected at 6, 12 and 24 hr after LPS inoculation and the number of infiltrating cells in the anterior chamber was counted. In addition, we assayed the concentration of protein, nitric oxide (NO), tumour necrosis factor-alpha (TNF-alpha) and prostaglandin E2 (PGE2). Immunohistochemical staining with a monoclonal antibody against activated NF-kappaB was performed in order to evaluate the effects of AST on NF-kappaB activation. Rats injected with AST showed a significant decrease in the number of infiltrating cells in the anterior chamber and additionally there was a significantly lower concentration of protein, NO, TNF-alpha and PGE2 in the aqueous humour. Moreover, even early stages of EIU were suppressed by injection of AST. The number of activated NF-kappaB-positive cells was lower in iris-ciliary bodies treated with 10 or 100 mg kg(-1) AST at 3 hr after LPS injection. These results suggest that AST reduces ocular inflammation in eyes with EIU by downregulating proinflammatory factors and by inhibiting the NF-kappaB-dependent signaling pathway.
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Adyuvantes Inmunológicos/uso terapéutico , Humor Acuoso/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Uveítis/tratamiento farmacológico , beta Caroteno/análogos & derivados , Animales , Humor Acuoso/inmunología , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/inmunología , Cuerpo Ciliar/metabolismo , Depresión Química , Dinoprostona/análisis , Ensayo de Inmunoadsorción Enzimática , Inmunohistoquímica/métodos , Iris/efectos de los fármacos , Iris/inmunología , Iris/metabolismo , Lipopolisacáridos , Masculino , Óxido Nítrico/análisis , Nitritos/análisis , Distribución Aleatoria , Ratas , Ratas Endogámicas Lew , Factor de Necrosis Tumoral alfa/análisis , Uveítis/inmunología , Uveítis/metabolismo , Xantófilas , beta Caroteno/uso terapéuticoRESUMEN
The aim of the present study was to investigate the efficacy of fucoxanthin on endotoxin-induced uveitis (EIU) in rats. The effects of fucoxanthin on endotoxin-induced leucocyte and protein infiltration, nitric oxide (NO), prostaglandin (PG)-E2 and tumour necrosis factor (TNF)-alpha concentrations in rat aqueous humour, as well as on the cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) protein expression in a mouse macrophage cell line (RAW 264.7 cells) were studied. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS injection, either 0.1, 1 or 10mgkg(-1) of fucoxanthin was injected intravenously. The aqueous humour was collected 24hr later from both eyes, and both the number of cells infiltrating into the aqueous humour and the aqueous humour protein concentration were measured. The levels of PGE2, NO and TNF-alpha were determined by enzyme-linked immunosorbent assay. The RAW 264.7 cells were pretreated with various concentrations of fucoxanthin for 24hr and subsequently incubated with LPS for 24hr. COX-2 and iNOS protein expression was analysed by the Western blotting method. Levels of PGE2, NO and TNF-alpha production were determined. Fucoxanthin suppressed the development of EIU in a dose-dependent fashion. Treatment with fucoxanthin resulted in a reduction in PGE2, NO and TNF-alpha concentrations in the aqueous humour. The expression of COX and iNOS protein in the fucoxanthin treated RAW264.7 cells decreased significantly compared to that the LPS group. It also significantly reduced the concentration of PGE2, NO and TNF-alpha production in the medium of cells. The present result indicate fucoxanthin suppresses the inflammation of EIU by blocking the iNOS and COX-2 protein expression and its anti-inflammatory effect on eye is comparable with the effect of predinisolone used in similar doses.
Asunto(s)
Uveítis Anterior/prevención & control , Xantófilas/uso terapéutico , Animales , Antiinflamatorios/uso terapéutico , Humor Acuoso/citología , Humor Acuoso/metabolismo , Western Blotting , Células Cultivadas , Medios de Cultivo , Ciclooxigenasa 2/metabolismo , Dinoprostona/metabolismo , Relación Dosis-Respuesta a Droga , Proteínas del Ojo/metabolismo , Lipopolisacáridos , Masculino , Ratones , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Prednisolona/uso terapéutico , Ratas , Ratas Endogámicas Lew , Factor de Necrosis Tumoral alfa/metabolismo , Uveítis Anterior/inducido químicamente , Uveítis Anterior/metabolismoRESUMEN
OBJECTIVE: To investigate the relationship between various factors and the undercorrection after laser in situ keratomileusis (LASIK). METHODS: Of 1 391 eyes (696 cases), the undercorrection eyes grouped by left or right eye were analyzed by Chi-square test and logistic regression analysis based on age, sex, occupation, address, family history of myopia and all ophthalmologic examination results before and after LASIK. RESULTS: The results of logistic regression analysis showed that the undercorrection after LASIK of both groups was related to four factors: the duration of myopia (left eye OR = 1.076, 95% CI: 1.030 - 1.124; right eye OR = 1.093, 95% CI: 1.046 - 1.142), preoperative refractive power (left eye OR = 7.799, 95% CI: 1.755 - 34.654; right eye OR = 28.823, 95% CI: 5.750 - 144.467), preoperative best corrected vision (left eye OR = 0.000, 95% CI: 0.000 - 0.262; right eye OR = 0.000, 95% CI: 0.000 - 0.144) and the corneal thickness (left eye OR = 0.976, 95% CI: 0.965 - 0.987; right eye OR = 0.975, 95% CI: 0.964 - 0.986). CONCLUSION: Longer duration of myopia and the more refractive power seemed to be the risk factors, while the better corrected vision and the thicker thickness of cornea the protect factors of undercorrection after LASIK.