Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 72
Filtrar
1.
Nat Commun ; 15(1): 3562, 2024 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-38670966

RESUMO

The diagnosis of inherited retinal degeneration (IRD) is challenging owing to its phenotypic and genotypic complexity. Clinical information is important before a genetic diagnosis is made. Metabolomics studies the entire picture of bioproducts, which are determined using genetic codes and biological reactions. We demonstrated that the common diagnoses of IRD, including retinitis pigmentosa (RP), cone-rod dystrophy (CRD), Stargardt disease (STGD), and Bietti's crystalline dystrophy (BCD), could be differentiated based on their metabolite heatmaps. Hundreds of metabolites were identified in the volcano plot compared with that of the control group in every IRD except BCD, considered as potential diagnosing markers. The phenotypes of CRD and STGD overlapped but could be differentiated by their metabolomic features with the assistance of a machine learning model with 100% accuracy. Moreover, EYS-, USH2A-associated, and other RP, sharing considerable similar characteristics in clinical findings, could also be diagnosed using the machine learning model with 85.7% accuracy. Further study would be needed to validate the results in an external dataset. By incorporating mass spectrometry and machine learning, a metabolomics-based diagnostic workflow for the clinical and molecular diagnoses of IRD was proposed in our study.


Assuntos
Aprendizado de Máquina , Metabolômica , Degeneração Retiniana , Retinose Pigmentar , Doença de Stargardt , Humanos , Metabolômica/métodos , Diagnóstico Diferencial , Degeneração Retiniana/diagnóstico , Degeneração Retiniana/sangue , Degeneração Retiniana/genética , Degeneração Retiniana/metabolismo , Masculino , Feminino , Retinose Pigmentar/diagnóstico , Retinose Pigmentar/genética , Retinose Pigmentar/sangue , Retinose Pigmentar/metabolismo , Doença de Stargardt/genética , Adulto , Pessoa de Meia-Idade , Adolescente , Adulto Jovem , Biomarcadores/sangue , Metaboloma , Criança , Distrofias de Cones e Bastonetes/diagnóstico , Distrofias de Cones e Bastonetes/genética , Distrofias de Cones e Bastonetes/sangue , Distrofias de Cones e Bastonetes/metabolismo , Espectrometria de Massas , Degeneração Macular/sangue , Degeneração Macular/diagnóstico , Degeneração Macular/genética
2.
Int J Mol Sci ; 23(1)2021 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-35008772

RESUMO

We investigate glial cell activation and oxidative stress induced by taurine deficiency secondary to ß-alanine administration and light exposure. Two months old Sprague-Dawley rats were divided into a control group and three experimental groups that were treated with 3% ß-alanine in drinking water (taurine depleted) for two months, light exposed or both. Retinal and external thickness were measured in vivo at baseline and pre-processing with Spectral-Domain Optical Coherence Tomography (SD-OCT). Retinal cryostat cross sections were immunodetected with antibodies against various antigens to investigate microglial and macroglial cell reaction, photoreceptor outer segments, synaptic connections and oxidative stress. Taurine depletion caused a decrease in retinal thickness, shortening of photoreceptor outer segments, microglial cell activation, oxidative stress in the outer and inner nuclear layers and the ganglion cell layer and synaptic loss. These events were also observed in light exposed animals, which in addition showed photoreceptor death and macroglial cell reactivity. Light exposure under taurine depletion further increased glial cell reaction and oxidative stress. Finally, the retinal pigment epithelial cells were Fluorogold labeled and whole mounted, and we document that taurine depletion impairs their phagocytic capacity. We conclude that taurine depletion causes cell damage to various retinal layers including retinal pigment epithelial cells, photoreceptors and retinal ganglion cells, and increases the susceptibility of the photoreceptor outer segments to light damage. Thus, beta-alanine supplements should be used with caution.


Assuntos
Luz , Neuroglia/patologia , Neuroglia/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Degeneração Retiniana/patologia , Taurina/metabolismo , Animais , Contagem de Células , Sobrevivência Celular , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Microglia/patologia , Neuroglia/metabolismo , Células Fotorreceptoras de Vertebrados/patologia , Ratos Sprague-Dawley , Degeneração Retiniana/sangue , Degeneração Retiniana/diagnóstico por imagem , Epitélio Pigmentado da Retina/diagnóstico por imagem , Epitélio Pigmentado da Retina/patologia , Sinapses/metabolismo , Taurina/sangue , Tomografia de Coerência Óptica , beta-Alanina
3.
Int J Mol Sci ; 21(20)2020 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-33050198

RESUMO

PURPOSE: The purpose of this study was to examine the effect of plasma rich in growth factors (PRGFs) under blue light conditions in an in vivo model of retinal degeneration. METHODS: Male Wistar rats were exposed to dark/blue light conditions for 9 days. On day 7, right eyes were injected with saline and left eyes with PRGF. Electroretinography (ERG) and intraocular pressure (IoP) measurements were performed before and after the experiment. After sacrifice, retinal samples were collected. Hematoxylin and eosin staining was performed to analyze the structure of retinal sections. Immunofluorescence for brain-specific homeobox/POU domain protein 3A (Brn3a), choline acetyltransferase (ChAT), rhodopsin, heme oxygenase-1 (HO-1), and glial fibrillary acidic protein (GFAP) was performed to study the retinal conditions. RESULTS: Retinal signaling measured by ERG was reduced by blue light and recovered with PRGF; however, IoP measurements did not show significant differences among treatments. Blue light reduced the expression for Brn3a, ChAT, and rhodopsin. Treatment with PRGF showed a recovery in their expressions. HO-1 and GFAP results showed that blue light increased their expression but the use of PRGF reduced the effect of light. CONCLUSIONS: Blue light causes retinal degeneration. PRGF mitigated the injury, restoring the functionality of these cells and maintaining the tissue integrity.


Assuntos
Biomarcadores , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Degeneração Retiniana/sangue , Degeneração Retiniana/etiologia , Animais , Biópsia , Sobrevivência Celular , Eletrorretinografia , Imunofluorescência , Imuno-Histoquímica , Pressão Intraocular , Luz , Ratos , Degeneração Retiniana/diagnóstico , Transdução de Sinais
4.
Sci Rep ; 10(1): 5255, 2020 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-32210247

RESUMO

Spectral-domain optical coherence tomography (SD-OCT) represents a reliable tool for retinal layer volume and thickness measurement. The aim of this study was to evaluate retinal changes indicating neurodegenerative processes in patients with end-stage renal disease (ESRD) compared to healthy controls. This was a cross-sectional, single-center study comprising 32 ESRD patients and 38 controls. Sectoral retinal nerve fiber layer (RNFL) thickness and retinal layer volumes were obtained by SD-OCT. Age- and gender-adjusted retinal layer volumes such as total retinal volume (p = 0.037), ganglion cell layer volume (GCL, p = 0.003), ganglion cell layer - inner plexiform layer volume (GCL-IPL, p = 0.005) and inner retinal layer volume (IRL, p = 0.042) of the right eye were lower in ESRD patients. Inner plexiform layer volume of both eyes (IPL, right eye: p = 0.017; left eye: 0.044) was reduced, as was RNFL thickness in the temporal superior sector (right eye: p = 0.016). A subgroup analysis excluding patients with diabetes revealed that GCL (p = 0.014) and GCL-IPL volume of the right eye (p = 0.024) and temporal superior sector of the RNFL scan (p = 0.021) in ESRD patients were still significantly thinner. We observed a decrease in several retinal layer volumes and temporal RNFL thickness indicative of retinal neurodegenerative processes in patients with ESRD.


Assuntos
Falência Renal Crônica/complicações , Nervo Óptico/diagnóstico por imagem , Retina/diagnóstico por imagem , Degeneração Retiniana/etiologia , Tomografia de Coerência Óptica/métodos , Estudos de Casos e Controles , Estudos Transversais , Nefropatias Diabéticas/complicações , Nefropatias Diabéticas/terapia , Retinopatia Diabética/complicações , Retinopatia Diabética/diagnóstico por imagem , Retinopatia Diabética/patologia , Feminino , Hemoglobinas Glicadas/análise , Hematócrito , Hemoglobinas/análise , Humanos , Falência Renal Crônica/patologia , Falência Renal Crônica/terapia , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Fibras Nervosas/ultraestrutura , Nervo Óptico/patologia , Diálise Renal , Retina/patologia , Degeneração Retiniana/sangue , Degeneração Retiniana/diagnóstico por imagem , Degeneração Retiniana/patologia , Células Ganglionares da Retina/patologia
5.
J Vet Intern Med ; 33(6): 2587-2594, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31660652

RESUMO

BACKGROUND: Sudden acquired retinal degeneration syndrome (SARDS) has clinical similarity to pituitary-dependent hypercortisolism (PDH) in dogs. Some studies have identified a greater frequency of SARDS in seasons with reduced daylight hours. Neurohormone imbalances contribute to retinal lesions in other species, warranting further study in dogs with SARDS. HYPOTHESIS: Dysregulation of circulating melatonin concentration is present in dogs with SARDS but not in dogs with PDH. ANIMALS: Fifteen client-owned dogs with spontaneous SARDS (median time of vision loss 18 days), 14 normal dogs, and 13 dogs with confirmed PDH. PROCEDURES: Prospective case-control study. ELISA on samples (obtained in the morning) for measurement of plasma melatonin and dopamine, serum serotonin, urine 6-sulfatoxymelatonin (MT6s), and creatinine. Statistical analysis was performed using 1-way ANOVA, Spearman correlation and receiver operator characteristic area under the curve analysis. RESULTS: There were no significant differences in circulating melatonin, serotonin or dopamine concentrations between the 3 groups, although the study was underpowered for detection of significant differences in serum serotonin. Urine MT6s:creatinine ratio was significantly higher in dogs with PDH (4.08 ± 2.15 urine [MT6s] ng/mL per mg of urine creatinine) compared with dogs with SARDS (2.37 ± .51, P < .01), but not compared with normal dogs. CONCLUSIONS AND CLINICAL RELEVANCE: We have identified neurohormone differences between dogs with SARDS and PDH.


Assuntos
Doenças do Cão/sangue , Dopamina/sangue , Melatonina/sangue , Hipersecreção Hipofisária de ACTH/veterinária , Degeneração Retiniana/veterinária , Serotonina/sangue , Animais , Estudos de Casos e Controles , Doenças do Cão/metabolismo , Cães , Melatonina/análogos & derivados , Melatonina/urina , Hipersecreção Hipofisária de ACTH/sangue , Hipersecreção Hipofisária de ACTH/metabolismo , Estudos Prospectivos , Degeneração Retiniana/sangue , Degeneração Retiniana/metabolismo
6.
Autophagy ; 15(1): 34-57, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30081747

RESUMO

ZFYVE26/Spastizin and SPG11/Spatacsin encode 2 large proteins that are mutated in hereditary autosomal-recessive spastic paraplegia/paraparesis (HSP) type 15 (AR-SPG15) and type 11 (AR-SPG11), respectively. We previously have reported that AR-SPG15-related ZFYVE26 mutations lead to autophagy defects with accumulation of immature autophagosomes. ZFYVE26 and SPG11 were found to be part of a complex including the AP5 (adaptor related protein complex 5) and to have a critical role in autophagic lysosomal reformation with identification of autophagic and lysosomal defects in cells with both AR-SPG15- and AR-SPG11-related mutations. In spite of these similarities between the 2 proteins, here we report that ZFYVE26 and SPG11 are differently involved in autophagy and endocytosis. We found that both ZFYVE26 and SPG11 interact with RAB5A and RAB11, 2 proteins regulating endosome trafficking and maturation, but only ZFYVE26 mutations affected RAB protein interactions and activation. ZFYVE26 mutations lead to defects in the fusion between autophagosomes and endosomes, while SPG11 mutations do not affect this step and lead to a milder autophagy defect. We thus demonstrate that ZFYVE26 and SPG11 affect the same cellular physiological processes, albeit at different levels: both proteins have a role in autophagic lysosome reformation, but only ZFYVE26 acts at the intersection between endocytosis and autophagy, thus representing a key player in these 2 processes. Indeed expression of the constitutively active form of RAB5A in cells with AR-SPG15-related mutations partially rescues the autophagy defect. Finally the model we propose demonstrates that autophagy and the endolysosomal pathway are central processes in the pathogenesis of these complicated forms of hereditary spastic paraparesis. Abbreviations: ALR, autophagic lysosome reformation; AP5, adaptor related protein complex 5; AR, autosomal-recessive; HSP, hereditary spastic paraplegia/paraparesis; ATG14, autophagy related 14; BafA, bafilomycin A1; BECN1, beclin 1; EBSS, Earle balanced salt solution; EEA1, early endosome antigen 1; EGF, epidermal growth factor; EGFR, epidermal growth factor receptor; GDP, guanosine diphosphate; GFP, green fluorescent protein; GTP, guanosine triphosphate; HSP, hereditary spastic paraplegias; LBPA, lysobisphosphatidic acid; MAP1LC3B/LC3B, microtubule associated protein 1 light chain 3 beta; MVBs, multivesicular bodies; PIK3C3, phosphatidylinositol 3-kinase, catalytic subunit type 3; PIK3R4, phosphoinositide-3-kinase regulatory subunit 4; PtdIns3P, phosphatidylinositol-3-phosphate; RFP, red fluorescent protein; RUBCN, RUN and cysteine rich domain containing beclin 1 interacting protein; shRNA, short hairpin RNA; SQSTM1/p62, sequestosome 1; TCC: thin corpus callosum; TF, transferrin; UVRAG, UV radiation resistance associated.


Assuntos
Autofagia/genética , Proteínas de Transporte/genética , Endocitose/genética , Proteínas/genética , Degeneração Retiniana/genética , Paraplegia Espástica Hereditária/genética , Proteínas Adaptadoras de Transporte Vesicular/sangue , Autofagossomos/metabolismo , Proteínas de Transporte/sangue , Proteínas de Transporte/metabolismo , Endossomos/metabolismo , Feminino , Células HeLa , Humanos , Lisossomos/metabolismo , Masculino , Mutação , Proteínas/metabolismo , Degeneração Retiniana/sangue , Paraplegia Espástica Hereditária/sangue , Proteínas rab de Ligação ao GTP/sangue , Proteínas rab de Ligação ao GTP/metabolismo , Proteína rab2 de Ligação ao GTP/sangue , Proteínas rab5 de Ligação ao GTP/sangue , Proteínas rab5 de Ligação ao GTP/genética , Proteínas rab5 de Ligação ao GTP/metabolismo
7.
J Immunol Methods ; 461: 44-52, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29772250

RESUMO

Ranibizumab (Lucentis®), a humanized antigen-binding fragment (Fab) monoclonal antibody that blocks VEGF-A activity, is currently approved for the treatment of several retinal degenerative diseases. The assessment of drug pharmacokinetics (PK) is essential for evaluating exposure as it relates to drug safety and efficacy. For drugs administered intravitreally, systemic drug levels during the course of clinical studies are typically 100 to 1000-fold lower than those of similar therapeutics dosed intravenously, posing a significant bioanalytical challenge for PK measurements. Thus, the development of a highly-sensitive assay for measuring pg/mL levels of ranibizumab in patients' sera after intravitreal administration was needed to support clinical studies. In this report, we describe the development of a novel method that utilizes a high-affinity murine monoclonal anti-ranibizumab-VEGF-complexes antibody (MARA) reagent to measure ranibizumab in human serum. The assay format utilizes a semi-homogeneous solution phase step using a monoclonal antibody (the MARA) that binds specifically to the ranibizumab-VEGF complex, but not to either alone. This unique reagent exhibited low non-specific binding and high selectivity, increasing signal-to-noise readouts and maximizing assay sensitivity. The resulting MARA enzyme-linked immunosorbent assay (ELISA) has a lower limit of quantification of 15 pg/mL in human serum. In the assay, serum samples are incubated overnight with a mixture containing biotinylated-VEGF and MARA, which form a three-molecule complex with ranibizumab in the sample. These complexes are then captured onto streptavidin-coated wells, followed by enzymatic detection using a horseradish peroxidase-labeled-anti-murine antibody reagent and a colorimetric reaction. The assay conditions were optimized to allow for quantitative detection of "total" ranibizumab levels in serum. The assay was fully validated, establishing its high tolerance to sample matrix, as well as its suitable specificity, accuracy, dilution linearity, as well as intra- and inter-assay precision. The MARA ELISA's novel and unique approach has resulted in a considerably more sensitive ranibizumab PK assay compared to earlier versions of this assay. The MARA ELISA has been used for PK measurements in multiple ranibizumab studies, supporting this drug's life-cycle management and related preclinical and clinical-development studies.


Assuntos
Anticorpos Monoclonais Murinos/química , Bevacizumab/farmacocinética , Degeneração Retiniana/sangue , Animais , Anticorpos Monoclonais Murinos/imunologia , Bevacizumab/imunologia , Bevacizumab/uso terapêutico , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Degeneração Retiniana/tratamento farmacológico , Degeneração Retiniana/imunologia , Fator A de Crescimento do Endotélio Vascular/sangue , Fator A de Crescimento do Endotélio Vascular/imunologia
8.
J Tissue Eng Regen Med ; 12(3): e1370-e1382, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-28715614

RESUMO

As the world's population begins to age, retinal degeneration is an increasing problem, and various treatment modalities are being developed. However, there have been no therapies for degenerative retinal conditions that are not characterized by neovascularization. We investigated whether transplantation of mouse adipose tissue-derived stem cells (mADSC) into the intraperitoneal space has a rescue effect on NaIO3 -induced retinal degeneration in mice. In this study, mADSC transplantation recovered visual function and preserved the retinal outer layer structure compared to the control group without any integration of mADSC into the retina. Moreover, endogenous ciliary neurotrophic factor (CNTF) was elevated in the retinas of mADSC-treated mice. We found that lipopolysaccharide (LPS) or LPS-stimulated monocyte supernatant induced the secretion of granulocyte colony stimulating factor (GCSF), CD54, CXCL10, interleukin-6 (IL-6), and CCL5 from the mADSC by cytokine array. Network inference was conducted to investigate signaling networks related to CNTF regulation. Based on bioinformatics data, the expression of IL-6 was related to the expression of CNTF. Additionally, intravitreal injection of IL-6 in rats produced up-regulation of endogenous CNTF in the retina. mADSC had a rescue effect on retinal degeneration through the up-regulation of endogenous CNTF by IL-6. Thus, transplantation of mADSC could be a potential treatment option for retinal degeneration.


Assuntos
Tecido Adiposo/citologia , Fator Neurotrófico Ciliar/metabolismo , Interleucina-6/metabolismo , Degeneração Retiniana/terapia , Transplante de Células-Tronco , Células-Tronco/citologia , Regulação para Cima , Animais , Células Cultivadas , Modelos Animais de Doenças , Injeções Intraperitoneais , Interleucina-6/sangue , Injeções Intravítreas , Iodatos , Lipopolissacarídeos/farmacologia , Camundongos Endogâmicos C57BL , Substâncias Protetoras/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Degeneração Retiniana/sangue , Degeneração Retiniana/genética , Degeneração Retiniana/patologia , Epitélio Pigmentado da Retina/patologia
9.
Invest Ophthalmol Vis Sci ; 58(13): 5985-5992, 2017 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-29183044

RESUMO

Purpose: The pathophysiology of neuroretinal thinning in children with human immunodeficiency virus (HIV) is poorly understood. The current study aimed to assess whether neuroretinal thinning in clinically stable perinatally HIV-infected children was associated with biomarkers of immune activation, inflammation, and neuronal damage. Methods: Inflammation-associated and neuronal damage markers were measured in blood and cerebrospinal fluid (CSF) of HIV-infected children aged 8 to 18 years. Using mixed-effects regression analyses, we assessed associations between these biomarkers and neuroretinal layer thickness, as measured with spectral-domain optical coherence tomography. Results: Thirty-two HIV-infected children (median age 13.6 years, 50% male) were included. Blood plasma levels of interleukin-6, monocyte chemoattractant protein-1, and soluble intercellular adhesion molecule-1 were inversely correlated with foveal inner plexiform layer thickness (coef = -4.40, P < 0.001; coef = -9.67, P = 0.047; coef = -10.48, P = 0.042, respectively). Plasma interleukin-6 was inversely correlated with foveal ganglion cell layer thickness (coef = -2.49, P = 0.010). Total Tau levels in CSF were inversely correlated with outer nuclear layer and inner segments thickness (foveal: coef = -19.3, P = 0.029; pericentral: coef = -18.09, P = 0.006) and pericentral total retinal thickness (coef = -28.2, P = 0.017). Conclusions: Neuroretinal thinning was associated with inflammation-associated and neuronal injury biomarkers in a cohort of antiretroviral therapy-treated perinatally HIV-infected children. These findings suggest that ongoing immune activation, inflammation, and neuronal injury occur in parallel with retinal thinning in pediatric HIV.


Assuntos
Infecções por HIV/complicações , Retina/patologia , Degeneração Retiniana , Adolescente , Biomarcadores/sangue , Biomarcadores/líquido cefalorraquidiano , Quimiocina CCL2/sangue , Criança , Feminino , Humanos , Molécula 1 de Adesão Intercelular/sangue , Interleucinas/sangue , Masculino , Análise de Regressão , Degeneração Retiniana/sangue , Degeneração Retiniana/líquido cefalorraquidiano , Degeneração Retiniana/patologia , Células Ganglionares da Retina/patologia , Tomografia de Coerência Óptica , Proteínas tau/líquido cefalorraquidiano
10.
J Diabetes Complications ; 31(5): 864-868, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28279572

RESUMO

AIM: To study the association of serum levels of anti-myeloperoxidase (MPO) antibody with retinal photoreceptor ellipsoid zone (EZ) disruption in diabetic retinopathy. METHODS: Consecutive patients with type 2 DM [diabetes mellitus with no retinopathy (NODR; n=20); non-proliferative diabetic retinopathy (NPDR; n=18); proliferative diabetic retinopathy (PDR; n=16)] and healthy controls (n=20) between the ages of 40 and 65years were included. Disruption of EZ was graded by spectral domain optical coherence tomography as no disruption of EZ and disrupted EZ. The serum levels of anti-MPO antibody was analyzed using standard protocol. Association between the variables was evaluated using multiple regression analysis. RESULTS: A significant difference was found between the serum levels of anti-MPO antibody in various study groups (p<0.001). A positive association was found between EZ disruption and levels of anti-MPO antibody [adjusted odd's ratio (AOR)=1.079, CI 1.010-1.124, p=0.04]. A significant positive correlation was found between logMAR visual acuity and grade of disruption (AOR=1.008, CI 1.006-5.688, p=0.04). CONCLUSIONS: An increased serum anti-MPO antibody levels is associated with retinal photoreceptor EZ disruption and decreased visual acuity in diabetic retinopathy.


Assuntos
Retinopatia Diabética/diagnóstico por imagem , Isoanticorpos/análise , Modelos Imunológicos , Peroxidase/antagonistas & inibidores , Células Fotorreceptoras de Vertebrados/patologia , Retina/diagnóstico por imagem , Degeneração Retiniana/complicações , Estudos Transversais , Diabetes Mellitus Tipo 2/complicações , Retinopatia Diabética/sangue , Retinopatia Diabética/imunologia , Feminino , Humanos , Imageamento Tridimensional , Índia , Masculino , Pessoa de Meia-Idade , Células Fotorreceptoras de Vertebrados/imunologia , Análise de Regressão , Reprodutibilidade dos Testes , Retina/imunologia , Degeneração Retiniana/sangue , Degeneração Retiniana/diagnóstico por imagem , Degeneração Retiniana/imunologia , Centros de Atenção Terciária , Tomografia de Coerência Óptica , Acuidade Visual
11.
Acta Diabetol ; 54(5): 489-497, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28238189

RESUMO

AIMS: Recent studies have identified neuroretinal abnormalities in persons affected by diabetes mellitus, before the onset of microvascular alterations. However, the role of glycemic variability (GV) on early retinal neurodegeneration is still not clarified. METHODS: To explore the relationship between glycemic control and neuroretinal characteristics, 37 persons with Type 1 diabetes mellitus (Type 1 DM) divided into two groups with no signs (noRD) and with mild non-proliferative diabetic retinopathy (NPDR) compared to 13 healthy control participants (C) were recruited. All persons underwent an optical coherence tomography with automatic segmentation of all neuroretinal layers. Measurements of mean of nasal (N)/temporal (T)/superior (S)/inferior (I) macular quadrants for individual layer were also calculated. Metabolic control was evaluated by glycated hemoglobin (HbA1c), and indexes of GV were calculated from continuous glucose monitoring. RESULTS: The difference among the three groups in terms of RNFL thickness was significantly dependent on quadrant (F(6;132) = 2.315; p = 0.037). This interaction was due to a specific difference in RNFL-N thickness, where both Type 1 DM groups showed a similar reduction versus C (-3.9 for noDR and -4.9 for NPDR), without any relevant difference between them (-1.0). Inner nuclear layer (INL) was increased in all quadrants in the two Type 1 DM groups compared to C (mean difference = 7.73; 95% CI: 0.32-15.14, p = 0.043; mean difference = 7.74; 95% CI: 0.33-15.15, p = 0.043, respectively). A negative correlation between RNFL-N and low blood glucose index (r = -0.382, p = 0.034) and positive correlation between INL and continuous overall net glycemic action -1, -2, -4 h (r = 0.40, p = 0.025; r = 0.39, p = 0.031; r = 0.41, p = 0.021, respectively) were observed in Type 1 DM patients. The triglycerides were positively and significantly correlated to INL (r = 0.48, p = 0.011), in Type 1 DM subjects. GV and triglycerides resulted both independent predictors of increased INL thickness. No correlation was found with HbA1c. CONCLUSIONS: Early structural damage of neuroretina in persons with Type 1 DM patients is related to glucose fluctuations. GV should be addressed, even in the presence of a good metabolic control.


Assuntos
Glicemia/metabolismo , Diabetes Mellitus Tipo 1/sangue , Retinopatia Diabética/sangue , Degeneração Retiniana/sangue , Adulto , Diabetes Mellitus Tipo 1/diagnóstico por imagem , Diabetes Mellitus Tipo 1/fisiopatologia , Retinopatia Diabética/diagnóstico por imagem , Retinopatia Diabética/fisiopatologia , Feminino , Índice Glicêmico/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Degeneração Retiniana/diagnóstico por imagem , Degeneração Retiniana/fisiopatologia , Tomografia de Coerência Óptica
12.
PLoS One ; 10(6): e0128395, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26042773

RESUMO

In the present study, we have evaluated one of the dietary supplements enriched with antioxidants and fish oil used in clinical care for patient with age-related macular degeneration. Rats were orally fed by a gastric canula daily with 0.2 ml of water or dietary supplement until they were sacrificed. After one week of treatment, animals were either sacrificed for lipid analysis in plasma and retina, or used for evaluation of rod-response recovery by electroretinography (ERG) followed by their sacrifice to measure rhodopsin content, or used for progressive light-induced retinal degeneration (PLIRD). For PLIRD, animals were transferred to bright cyclic light for one week. Retinal damage was quantified by ERG, histology and detection of apoptotic nuclei. Animals kept in dim-cyclic-light were processed in parallel. PLIRD induced a thinning of the outer nuclear layer and a reduction of the b-wave amplitude of the ERG in the water group. Retinal structure and function were preserved in supplemented animals. Supplement induced a significant increase in omega-3 fatty acids in plasma by 168% for eicosapentaenoic acid (EPA), 142% for docosapentaenoic acid (DPA) and 19% for docosahexaenoic acid (DHA) and a decrease in the omega-6 fatty acids, DPA by 28%. In the retina, supplement induced significant reduction of linolenic acid by 67% and an increase in EPA and DPA by 80% and 72%, respectively, associated with significant decrease in omega-6 DPA by 42%. Supplement did not affect rhodopsin content or rod-response recovery. The present data indicate that supplement rapidly modified the fatty acid content and induced an accumulation of EPA in the retina without affecting rhodopsin content or recovery. In addition, it protected the retina from oxidative stress induced by light. Therefore, this supplement might be beneficial to slow down progression of certain retinal degeneration.


Assuntos
Antioxidantes/uso terapêutico , Suplementos Nutricionais , Progressão da Doença , Ácidos Graxos Ômega-3/uso terapêutico , Luz/efeitos adversos , Degeneração Retiniana/patologia , Degeneração Retiniana/prevenção & controle , Animais , Apoptose/efeitos da radiação , Vias Biossintéticas/efeitos da radiação , Eletrorretinografia , Ácidos Graxos Ômega-3/sangue , Ácidos Graxos Ômega-6/sangue , Feminino , Masculino , Fármacos Neuroprotetores/uso terapêutico , Plasmalogênios/metabolismo , Ratos Sprague-Dawley , Regeneração/efeitos da radiação , Retina/patologia , Retina/efeitos da radiação , Degeneração Retiniana/sangue , Células Fotorreceptoras Retinianas Bastonetes/patologia , Células Fotorreceptoras Retinianas Bastonetes/efeitos da radiação , Rodopsina/metabolismo
13.
Mol Cell Biol ; 35(16): 2771-89, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26055327

RESUMO

Serum retinol-binding protein 4 (RBP4) is the sole specific transport protein for retinol in the blood, but it is also an adipokine with retinol-independent, proinflammatory activity associated with obesity, insulin resistance, type 2 diabetes, and cardiovascular disease. Moreover, two separate studies reported that patients with proliferative diabetic retinopathy have increased serum RBP4 levels compared to patients with mild or no retinopathy, yet the effect of increased levels of RBP4 on the retina has not been studied. Here we show that transgenic mice overexpressing RBP4 (RBP4-Tg mice) develop progressive retinal degeneration, characterized by photoreceptor ribbon synapse deficiency and subsequent bipolar cell loss. Ocular retinoid and bisretinoid levels are normal in RBP4-Tg mice, demonstrating that a retinoid-independent mechanism underlies retinal degeneration. Increased expression of pro-interleukin-18 (pro-IL-18) mRNA and activated IL-18 protein and early-onset microglia activation in the retina suggest that retinal degeneration is driven by a proinflammatory mechanism. Neither chronic systemic metabolic disease nor other retinal insults are required for RBP4 elevation to promote retinal neurodegeneration, since RBP4-Tg mice do not have coincident retinal vascular pathology, obesity, dyslipidemia, or hyperglycemia. These findings suggest that elevation of serum RBP4 levels could be a risk factor for retinal damage and vision loss in nondiabetic as well as diabetic patients.


Assuntos
Retina/patologia , Degeneração Retiniana/genética , Degeneração Retiniana/patologia , Retinoides/análise , Proteínas Plasmáticas de Ligação ao Retinol/genética , Envelhecimento , Animais , Humanos , Inflamação/genética , Inflamação/patologia , Interleucina-18/genética , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Retina/metabolismo , Degeneração Retiniana/sangue , Proteínas Plasmáticas de Ligação ao Retinol/análise , Sinapses/genética , Sinapses/patologia , Regulação para Cima
14.
Ophthalmic Surg Lasers Imaging Retina ; 46(3): 362-8, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25856824

RESUMO

BACKGROUND AND OBJECTIVE: To characterize the cellular, immunological, and inflammatory response to retinal photocoagulation of intense rupture laser lesions as a model of retinal degenerative diseases. MATERIALS AND METHODS: Seven C57BL/6 mice were irradiated using a 532-nm laser to induce 10 retinal burns per eye that ruptured Bruch's membrane. Blood was drawn from the saphenous vein before and 2 months after laser treatment. The serum was run on antigen microarrays with 85 molecular markers associated with retinal degenerative diseases. RESULTS: Rupture laser resulted in dramatic changes in the immunoglobulin reactivity of most inflammatory markers 2 months after laser injury. Approximately two-thirds increased expression and one-third decreased expression. Notable markers that were increased included complement C3, CRP, PKM2, and aldolase. CONCLUSION: Rupture laser injury causes a change in the serum inflammatory markers after 2 months similar to macular degeneration, diabetic retinopathy, and cancer-associated retinopathy. This animal model could be used as a biomarker for disease stage and activity in retinal degenerations.


Assuntos
Biomarcadores/sangue , Lâmina Basilar da Corioide/lesões , Modelos Animais de Doenças , Fotocoagulação a Laser/efeitos adversos , Degeneração Retiniana/sangue , Animais , Proteína C-Reativa/metabolismo , Complemento C3/metabolismo , Frutose-Bifosfato Aldolase/sangue , Imunoglobulina G/sangue , Inflamação , Camundongos , Camundongos Endogâmicos C57BL , Piruvato Quinase/sangue , Degeneração Retiniana/etiologia , Degeneração Retiniana/patologia , Ruptura , Veia Safena
15.
Acta Diabetol ; 51(2): 211-8, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23771613

RESUMO

Vasoregression characterizes diabetic retinopathy in animal models and in humans. We have recently demonstrated that vasoregression is earlier initiated in a rat model of ciliopathy-induced retinal neurodegeneration (TGR rat). The aim was to assess the balance between vasoregressive effects of chronic hyperglycemia and photoreceptor degeneration on adult vascular remodelling. The retinas were analyzed at 4 and 9 months after streptozotocin-induced diabetes. Neurodegeneration was determined by quantitation of cell numbers and retinal layer thickness. Vasoregression was assessed by quantitative retinal morphometry in retinal digest preparations. Retinal VEGF levels were measured by ELISA. Glial activation, expression and location of HSP27 and phosphorylated HSP27 were evaluated by immunofluorescence staining. Unexpectedly, the numbers of acellular capillaries were reduced at both time points and led to fewer intraretinal microvascular abnormalities in late stage diabetic TGR. Concomitantly, inner nuclear layers (INLs) in diabetic TGR rats were protected from cell loss at both time points. Consequently, glial activation was reduced, but VEGF level was increased in diabetic TGR retinas. Expressions of HSP27 were upregulated in glia cells in the preserved INL of diabetic TGR. Chronic hyperglycemia preserves the microvasculature in the retinal model of neurodegeneration. Cell preservation in the retinal INL was associated with protective gene regulation. Together, these data indicate that diabetes can induce vasoprotection, in which retinal glia can play a particular role.


Assuntos
Diabetes Mellitus Experimental/patologia , Retinopatia Diabética/sangue , Hiperglicemia/metabolismo , Degeneração Retiniana/sangue , Doenças Vasculares/sangue , Animais , Diabetes Mellitus Experimental/sangue , Feminino , Proteínas de Choque Térmico HSP27/sangue , Histocitoquímica , Humanos , Microscopia Confocal , Microscopia de Fluorescência , Neuroglia/patologia , Distribuição Aleatória , Ratos , Ratos Transgênicos , Doenças Vasculares/patologia , Fator A de Crescimento do Endotélio Vascular/sangue
16.
Mol Vis ; 19: 812-21, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23592918

RESUMO

PURPOSE: Diabetic retinopathy (DR) is a leading cause of vision loss in working-age people. To retard the development and progression of retina lesions, effective therapeutic strategies directed toward key molecular targets are desired. Phlorizin is effective in treating diabetic complications, but little is known about functional protein changes that may mediate its actions. The aim of this study was to identify retinal proteomic alterations in db/db mice treated with phlorizin. METHODS: We used C57BLKS/J db/db mice as a type 2 diabetic animal model, while C57BLKS/J db/m mice were selected as the control. Phlorizin (20 mg/kg bodyweight /d) was administrated to db/db mice for ten weeks. Serum fasting blood glucose and advanced glycation end products were determined. Meanwhile, retina cell apoptosis was determined with terminal transferase dUTP nick end labeling. Isobaric tags for relative and absolute quantification and subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to identify and profile retinal proteins among control, untreated diabetic, and phlorizin-treated db/db mice. The expression of glial fibrillary acidic protein was measured in retinas using western blotting analysis. RESULTS: Phlorizin treatment significantly reduced fasting blood glucose and levels of advanced glycation end products (p<0.05) and remarkably inhibited retina cell apoptosis and the expression of glial fibrillary acidic protein in the retinas of db/db mice. In addition, we identified 1,636 proteins from retina tissue in total, of which 348 proteins were differentially expressed in db/db mice compared with the controls. Only 60 proteins in the retinas of the db/db mice were found to be differentially changed following phlorizin treatment, including 33 proteins that were downregulated and 27 proteins that were upregulated. Most of these differentially changed proteins were involved in oxidative stress, apoptosis, energy metabolism, and signaling transduction. CONCLUSIONS: Our study revealed the expression of proteins differentially changed after phlorizin therapy. These proteins are most likely to participate in the development and recovery of DR. Our findings help expand understanding of the mechanism underlying the onset and progression of DR, and provide novel targets for evaluating the effects of phlorizin therapy.


Assuntos
Retinopatia Diabética/tratamento farmacológico , Retinopatia Diabética/metabolismo , Marcação por Isótopo/métodos , Florizina/uso terapêutico , Proteômica/métodos , Animais , Apoptose/efeitos dos fármacos , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Western Blotting , Peso Corporal/efeitos dos fármacos , Biologia Computacional , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Retinopatia Diabética/patologia , Proteínas do Olho/metabolismo , Jejum/sangue , Proteína Glial Fibrilar Ácida/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Marcação In Situ das Extremidades Cortadas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Neuroglia/patologia , Florizina/farmacologia , Degeneração Retiniana/sangue , Degeneração Retiniana/patologia , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismo
17.
Curr Eye Res ; 38(6): 680-4, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23317171

RESUMO

PURPOSE: To compare peripapillary retinal nerve fiber layer (RNFL) thicknesses measured by Cirrus HD optical coherence tomography (OCT) of patients with vitamin B12 deficiency with healthy controls and to evaluate the correlation between the peripapillary RNFL thickness and plasma vitamin B12 levels. MATERIALS AND METHODS: Forty-five patients (19 male and 26 female) with a diagnosis of vitamin B12 deficiency (patient group) and 45 age- and sex- matched healthy subjects (control group) were consecutively enrolled in this study. Average, temporal, nasal, inferior, and superior quadrant peripapillary RNFL thicknesses of each subject were obtained using the Cirrus HD OCT. Disc area (DA) and rim area (RA), central subfield thickness (CST), cube volume (CV), and cube average thickness (CAT) were also measured. RESULTS: Mean age of each group was 33.1 ± 6.5 years (range: 21-45 years). Mean plasma vitamin B12 level was 114.8 ± 34.0 pg/mL in the patient group and was 405.1 ± 20.0 pg/mL in the control group (p < 0.001). The patient and control groups were similar regarding axial length, plasma folate levels, DA, RA, CST, CV, CAT, and RNFL thicknesses in superior, nasal, and inferior quadrants. However, average RNFL and RNFL in temporal quadrant were significantly thinner in the patient group than in the control group (p = 0.013 and p < 0.001, respectively). In addition, temporal (r = 0.356, p = 0.001) and average (r = 0.212, p = 0.045) peripapillary RNFL thicknesses were correlated with plasma vitamin B12 levels. CONCLUSION: We have shown that, as in other non-glaucomatous optic neuropathies, temporal quadrant RNFL thickness was thinner in patients with vitamin B12 deficiency and it was correlated with plasma vitamin B12 levels. Further studies are warranted to clarify the clinical relevance of these findings and the effects of vitamin B12 replacement therapy.


Assuntos
Fibras Nervosas/patologia , Doenças do Nervo Óptico/patologia , Retina/patologia , Degeneração Retiniana/patologia , Tomografia de Coerência Óptica/métodos , Deficiência de Vitamina B 12/patologia , Adulto , Feminino , Glaucoma/sangue , Glaucoma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Disco Óptico/patologia , Doenças do Nervo Óptico/sangue , Degeneração Retiniana/sangue , Vitamina B 12/sangue , Deficiência de Vitamina B 12/sangue , Adulto Jovem
18.
Cell Biochem Funct ; 31(1): 65-74, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22915345

RESUMO

Retinopathy, a common complication of diabetes, is characterized by an unbalanced production of nitric oxide (NO), a process regulated by nitric oxide synthase (NOS). We hypothesized that retinopathy might stem from changes in the insulin receptor substrate (IRS)/PI3K/AKT pathway and/or expression of NOS isoforms. Thus, we analysed the morphology and apoptosis index in retinas of obese rats in whom insulin resistance had been induced by a high-fat diet (HFD). Immunoblotting analysis revealed that the retinal tissue of HFD rats had lower levels of AKT(1) , eNOS and nNOS protein than those of samples taken from control animals. Furthermore, immunohistochemical analyses indicated higher levels of iNOS and 4-hydroxynonenal and a larger number of apoptotic nuclei in HFD rats. Finally, both the inner and outer retinal layers of HFD rats were thinner than those in their control counterparts. When considered alongside previous results, these patterns suggest two major ways in which HFD might impact animals: direct activity of ingested fatty acids and/or via insulin-resistance-induced changes in intracellular pathways. We discuss these possibilities in further detail and advocate the use of this animal model for further understanding relationships between retinopathy, metabolic syndrome and type 2 diabetes.


Assuntos
Gorduras na Dieta/toxicidade , Proteínas do Olho/fisiologia , Obesidade/fisiopatologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Degeneração Retiniana/etiologia , Animais , Apoptose , Astrócitos/patologia , Glicemia/análise , Retinopatia Diabética , Modelos Animais de Doenças , Ácidos Graxos/sangue , Proteínas Substratos do Receptor de Insulina/fisiologia , Resistência à Insulina , Peroxidação de Lipídeos , Lipídeos/sangue , Fígado/patologia , Masculino , Óxido Nítrico Sintase Tipo I/fisiologia , Óxido Nítrico Sintase Tipo III/fisiologia , Obesidade/sangue , Obesidade/complicações , Fosfatidilinositol 3-Quinases/fisiologia , Ratos , Ratos Wistar , Retina/metabolismo , Retina/patologia , Degeneração Retiniana/sangue , Degeneração Retiniana/fisiopatologia , Transdução de Sinais
19.
J Neuroimmunol ; 254(1-2): 91-100, 2013 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-23110938

RESUMO

Autoimmunity may contribute to retinal degeneration. The studies examined the evolution of autoimmune responses against retina in naïve dystrophic RCS rats over the course of retinal degeneration. We showed that anti-retinal autoantibodies and T cells are generated in response to the availability of antigenic material released from dying photoreceptor cells during retinal degeneration but with distinctive activation trends. Passive transfer of anti-retinal antibodies enhanced disease progression by disrupting the BRB, upregulating MCP-1, attracting blood macrophages into retina, and augmenting apoptotic photoreceptor cell death. Our findings directly link anti-retinal autoantibodies to activated macrophage entry and their possible role in neurodegeneration.


Assuntos
Arrestina/imunologia , Autoimunidade/fisiologia , Proteínas do Olho/imunologia , Macrófagos/patologia , Retina/patologia , Degeneração Retiniana , Proteínas de Ligação ao Retinol/imunologia , Transferência Adotiva/métodos , Fatores Etários , Análise de Variância , Animais , Anticorpos/farmacologia , Antígenos CD/metabolismo , Autoimunidade/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quimiocina CCL2/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Marcação In Situ das Extremidades Cortadas , Linfócitos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Masculino , Ratos , Ratos Mutantes , Retina/efeitos dos fármacos , Degeneração Retiniana/sangue , Degeneração Retiniana/imunologia , Degeneração Retiniana/patologia , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
20.
Exp Eye Res ; 97(1): 137-47, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22251455

RESUMO

Our purpose was to find a method to create a large animal model of inducible photoreceptor damage. To this end, we tested in domestic swine the efficacy of two chemical toxins, known to create photoreceptor damage in other species: Iodoacetic Acid (IAA) and Sodium Iodate (NaIO(3)). Intravenous (IV) administration of NaIO(3) up to 90 mg/kg had no effect on retinal function and 110 mg/kg was lethal. IV administration of IAA (5-20 mg/kg) produced concentration-dependent changes in visual function as measured by full-field and multi-focal electroretinograms (ffERG and mfERG), and 30 mg/kg IAA was lethal. The IAA-induced effects measured at two weeks were stable through eight weeks post-injection, the last time point investigated. IAA at 7.5, 10, and 12 mg/kg produce a concentration-dependent reduction in both ffERG b-wave and mfERG N1-P1 amplitudes compared to baseline at all post-injection times. Comparisons of dark- and light-adapted ffERG b-wave amplitudes show a more significant loss of rod relative to cone function. The fundus of swine treated with ≥10 mg/kg IAA was abnormal with thinner retinal vessels and pale optic discs, and we found no evidence of bone spicule formation. Histological evaluations show concentration-dependent outer retinal damage that correlates with functional changes. We conclude that NaIO(3,) is not an effective toxin in swine. In contrast, IAA can be used to create a rapidly inducible, selective, stable and concentration-dependent model of photoreceptor damage in swine retina. Because of these attributes this large animal model of controlled photoreceptor damage should be useful in the investigation of treatments to replace damaged photoreceptors.


Assuntos
Modelos Animais de Doenças , Inibidores Enzimáticos/toxicidade , Iodatos/toxicidade , Ácido Iodoacético/toxicidade , Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Degeneração Retiniana/induzido quimicamente , Animais , Glicemia/metabolismo , Adaptação à Escuridão , Relação Dose-Resposta a Droga , Eletrorretinografia , Infusões Intravenosas , Estimulação Luminosa , Células Fotorreceptoras de Vertebrados/patologia , Degeneração Retiniana/sangue , Degeneração Retiniana/fisiopatologia , Sus scrofa
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...