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The thorough degeneration of organelles in the core of the lens is certainly a hallmark event during the lens development. Organelles degradation in the terminal differentiation process of lens fiber cells to form an organelle-free zone is critical for lens maturation and transparency. Several mechanisms have been proposed to expand our understanding of lens organelles degradation, including apoptotic pathways, the participation of ribozyme, proteolytic enzyme and phospholipase A and acyltransferase, and the newly discovered roles for autophagy. Autophagy is a lysosome-dependent degradation reaction during which the "useless" cellular components are degraded and recycled. These cellular components, such as incorrectly folded proteins, damaged organelles and other macromolecules, are first engulfed by the autophagosome before being further delivered to lysosomes for degradation. Although autophagy has been recognized involving in organelle degradation of the lens, the detailed functions remain to be discovered. Recent advances have revealed that autophagy not only plays a vital role in the intracellular quality control of the lens but is also involved in the degradation of nonnuclear organelles in the process of lens fiber cell differentiation. Herein, we first review the potential mechanisms of organelle-free zone formation, then discuss the roles of autophagy in intracellular quality control and cataract formation, and finally substantially summarize the potential involvement of autophagy in the development of organelle-free zone formation.
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Catarata , Cristalino , Humanos , Orgánulos/metabolismo , Cristalino/metabolismo , Autofagia , Catarata/metabolismo , Lisosomas , Proteínas/metabolismoRESUMEN
PURPOSE: A self-controlled study to determine the influence of illuminance and correlated colour temperature (CCT) of light-emitting diode (LED) lighting on asthenopia. METHODS: Twenty-two healthy postgraduates (nine women) were recruited to read under eight LED lighting conditions with four illuminances (300 lx, 500 lx, 750 lx and 1000 lx) and four CCTs (2700, 4000, 5000 and 6500 K) for 2 h. A subjective asthenopia questionnaire, the optical quality analysis system (OQAS) and an inflammatory cytokine assay were used to assess the levels of asthenopia. RESULTS: Increased asthenopia was observed after reading, but the degree varied with lighting conditions. There were significant differences among the groups in terms of subjective symptoms (inattention, eye pain, dry eye and total score), optical performance parameters (modulation transfer function [MTF] cut-off frequency, Strehl ratio [SR], objective scattering index [OSI], mean OSI and accommodative amplitude [AA]) as well as inflammatory cytokines in the tears (epidermal growth factor [EGF], transforming growth factor [TGF]-α, interleukin [IL]-6, IL-8, macrophage inflammatory protein [MIP]-1ß, tumour necrosis factor [TNF]-α, TNF-ß and vascular endothelial growth factor [VEGF]-A). All of the subjective and objective measurements collectively suggested that asthenopia was lessened for the 500 lx-4000 K condition. However, asthenopia was significantly worse for 300 lx-2700 K and 1000 lx-6500 K in terms of subjective symptoms and objective optical performance, respectively. CONCLUSIONS: LED illuminance and CCT do have a significant effect on asthenopia during reading. 500 lx-4000 K lighting resulted in the lowest level of asthenopia. Conversely, low illuminance at low CCT (300 lx-2700 K) and high illuminance at high CCT (1000 lx-6500 K) promoted more severe asthenopia.
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Lectura , Factor A de Crecimiento Endotelial Vascular , Femenino , HumanosRESUMEN
Oxidative stress and endoplasmic reticulum (ER) stress are the key contributing factors for cataract progression. In our previous studies, we demonstrated that the nuclear factor erythroid 2-like-2 (Nrf-2)/heme oxygenase-1 (HO-1)/carbon monoxide (CO) axis protects lens epithelial cells (LECs) against oxidants and ER stress. In the present study, transgenic FVB/N mice overexpressing the negative dominant mutant HO-1 G143H (TgHO-1 G143H) were generated to evaluate the crosstalk among HO-1, oxidative stress and ER stress in maintaining lens transparency. Slit-lamp examination revealed that nuclear cataracts occurred at 4 months in the TgHO-1 G143H mice, which was 5 months earlier than that of the control mice. The lenses of the transgenic mice showed an accumulation of malondialdehyde and protein carbonyl with a decrease in glutathione and protein sulfhydryl levels. Elevated concentrations of ER stress biomarkers (Bip, PERK, ATF6, IRE1, CHOP, caspase-12 and caspase-3) in the lenses of the TgHO-1 G143H mice were identified by western blotting. Furthermore, we confirmed that overexpressed HO-1 G143H in LECs resulted in oxidative insult and apoptosis in vitro. All of these data suggested that HO-1 enzymatic activity loss induces early-onset nuclear cataracts by activating oxidative stress and ER stress.
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Catarata/patología , Estrés del Retículo Endoplásmico , Hemo-Oxigenasa 1/metabolismo , Estrés Oxidativo , Envejecimiento/patología , Animales , Apoptosis , Biomarcadores/metabolismo , Proliferación Celular , Células Epiteliales/metabolismo , Espacio Intracelular/metabolismo , Cristalino/patología , Ratones Mutantes , Ratones Transgénicos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND: This study aimed to identify and evaluate potential molecular targets associated with the development of proliferative diabetic retinopathy (DR). METHODS: The microarray dataset "GSE60436" generated from fibrovascular membranes (FVMs) associated with proliferative DR was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) from the active FVMs and control or inactive FVMs and control were evaluated and co-DEGs were identified using VEEN analysis. Functional enrichment analysis, and protein-protein interactions (PPI) network and module analyses were performed on the upregulated and downregulated coDEGs. Finally, several predictions regarding microRNAs (miRNAs) and transcription factors (TFs) were made to construct a putative TF-miRNA-target network. RESULTS: A total of 1475 co-DEGs were screened in active/inactive FVM samples, including 461 upregulated and 1014 downregulated genes, which were enriched for angiogenesis [Hypoxia Inducible Factor 1 Subunit Alpha (HIF1A) and Placental Growth Factor (PGF)] and visual perception, respectively. In the case of the upregulated co-DEGs, Kinesin Family Member 11 (KIF11), and BUB1 Mitotic Checkpoint Serine/Threonine Kinase (BUB1) exhibited the highest values in both the PPI network and module analyses, as well as the genes related to mitosis. In the case of downregulated co-DEGs, several G protein subunits, including G Protein Subunit Beta 3 (GNB3), exhibited the highest values in both the PPI network and module analyses. The genes identified in the module analysis were found to be from the signal transduction-related pathways. In addition, we were able to identify four miRNAs and five TFs, including miR-136 and miR-374. CONCLUSIONS: In brief, HIF1A, PGF, KIF11, G protein subunits, and miR-136, miR-374 may all be involved in angiogenesis, retinal endothelial cell proliferation, and visual signal transduction in proliferative DR. This study provides a number of novel insights that may aid the development of future studies dedicated to discovering novel therapeutic targets in proliferative DR.
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Retinopatía Diabética/genética , Células Endoteliales/patología , Perfilación de la Expresión Génica , Neovascularización Retiniana/genética , Proliferación Celular , Conjuntos de Datos como Asunto , Proteínas de Unión al GTP/genética , Redes Reguladoras de Genes , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Cinesinas/genética , MicroARNs/genética , Análisis por Micromatrices , Terapia Molecular Dirigida , Factor de Crecimiento Placentario/genética , Dominios y Motivos de Interacción de Proteínas/genética , Factores de Transcripción/genética , Visión Ocular/genéticaRESUMEN
Lens epithelial cells (LECs) play a critical role in the maintenance of clear crystalline lens. Previously, we reported that heme oxygenase-1 can protect LECs from hydrogen peroxide (H2O2)-induced apoptosis and oxidative stress; however, to the best of our knowledge, these protection mechanisms have not yet been explained. As carbon monoxide (CO) is an active by-product of heme degradation, we investigated its cytoprotective mechanism in both H2O2-treated human LECs (SRA 01/04) and primary rabbit LECs. CO-releasing molecule-3 was used as a CO releasing vehicle. The nuclear translocation of nuclear factor kappa B (NF-κB) p65 was monitored by Western blot and immunofluorescence staining. In addition, the levels of intracellular reactive oxygen species (ROS), antioxidants, and apoptotic molecules (Bax, Bcl-2, and caspase-3) were measured. Furthermore, cell apoptosis rate was quantified by flow cytometry. Our results disclosed that low concentrations of CO released from CO-releasing molecule-3 can attenuate NF-κB p65 nuclear translocation, reduce ROS generation, and enhance intracellular glutathione and superoxide dismutase levels. Moreover, low concentrations of CO inhibited H2O2-induced apoptotic molecules, thereby decreasing the apoptosis of LECs. These findings suggest that low concentrations of CO protect LECs from H2O2-induced oxidative damage by attenuating NF-κB p65 nuclear translocation, reducing the generation of ROS and apoptotic molecules, and restoring antioxidant enzyme levels, thereby inhibiting LECs apoptosis.
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Monóxido de Carbono/farmacología , Células Epiteliales/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Cristalino/metabolismo , FN-kappa B/metabolismo , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Antioxidantes/metabolismo , Apoptosis/efectos de los fármacos , Monóxido de Carbono/metabolismo , Caspasa 3 , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Células Cultivadas , Células Epiteliales/metabolismo , Humanos , Cristalino/citología , Conejos , Especies Reactivas de Oxígeno/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Mitochondrial components are of great importance for the maintenance of lens transparency. In our previous work, we confirmed that carbon monoxide (CO) can protect human and rabbit lens epithelial cells (LECs) from hydrogen peroxide (H2O2)-mediated apoptosis, while the mechanism remains undefined. Because CO can bind to mitochondrial cytochrome c oxidase (COX), we evaluated the effect of CO on the regulation of mitochondrial biogenesis and function in H2O2-treated rabbit LECs. To evaluate mitochondrial biogenesis, several mitochondrial transcription factors (PGC-1α, NRF-1, and mtTFA) were detected by western blot analysis. To assess cellular metabolism, adenosine triphosphate (ATP) levels and COX enzymatic activity were measured. In addition, mitochondrial permeability transition pores (mPTP) opening, dissipation of mitochondrial membrane potential (ΔΨm), cytochrome c mitochondrial translocation, and apoptotic molecules were also detected to evaluate mitochondrial apoptosis pathway. Furthermore, the interaction of Bcl-2 and COX was assessed by co-immunoprecipitation. Finally, CO-mediated regulation of cellular function was detected in Bcl-2-knockdown cells. Our results confirmed that CO pretreatment restored H2O2-induced down-regulation of mitochondrial transcription factors expression, COX activity and ATP production. Moreover, CO pretreatment attenuated mPTP opening, ΔΨm loss, cytochrome c mitochondrial translocation, and activation of apoptotic molecules. Bcl-2 was identified to bind to COX, and silence of Bcl-2 expression prevented CO-regulated cellular metabolism and cytoprotection. These data suggest that CO modulates H2O2-induced cellular dysfunction by increasing mitochondrial biogenesis, enhancing cellular metabolism, and attenuating mitochondrial apoptosis cascade. Moreover, Bcl-2 expression was vital for CO to regulate cellular metabolism and cytoprotection in LECs.
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Antimetabolitos/farmacología , Monóxido de Carbono/farmacología , Células Epiteliales/efectos de los fármacos , Peróxido de Hidrógeno/toxicidad , Cristalino/citología , Mitocondrias/efectos de los fármacos , Oxidantes/toxicidad , Adenosina Trifosfato/metabolismo , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Proteínas de Unión al ADN/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Células Epiteliales/metabolismo , Potencial de la Membrana Mitocondrial/fisiología , Mitocondrias/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Poro de Transición de la Permeabilidad Mitocondrial , Proteínas Mitocondriales/metabolismo , Factor 1 Relacionado con NF-E2/metabolismo , Biogénesis de Organelos , Compuestos Organometálicos/farmacología , Estrés Oxidativo/efectos de los fármacos , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Conejos , Factores de Transcripción/metabolismoRESUMEN
Our previous study has shown heme oxygenase-1 (HO-1) protects human lens epithelial cells (LECs) against H2O2-induced oxidative stress and apoptosis. Nrf2, the major regulator of HO-1, is triggered during the mutual induction of oxidative stress and ER stress. In response to ER stress, unfolded protein response (UPR) serves as a program of transcriptional and translational regulation mechanism with PERK involved. Both Nrf2 and ATF4 are activated as the downstream effect of PERK signaling coordinating the convergence of dual stresses. However, the ways in which Nrf2 interacting with ATF4 regulates deteriorated redox state have not yet been fully explored. Here, the transfected LECs with Nrf2 overexpression illustrated enhanced resistance in morphology and viability upon H2O2 treatment condition. Intracellular ROS accumulation arouses ER stress, initiating PERK dependent UPR and inducing the downstream signal Nrf2 and ATF4 auto-phosphorylation. Further, converging at target promoters, ATF4 facilitates Nrf2 with the expression of ARE-dependent phase II antioxidant and detoxification enzymes. According to either Nrf2 or ATF4 gene modification, our data suggests a novel interaction between Nrf2 and ATF4 under oxidative and ER stress, thus drives specific enzymatic and non-enzymatic reactions of antioxidant mechanisms maintaining redox homeostasis. Therapies that restoring Nrf2 or ATF4 expression might help to postpone LECs aging and age-related cataract formation.
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Factor de Transcripción Activador 4/metabolismo , Estrés del Retículo Endoplásmico , Células Epiteliales/citología , Cristalino/citología , Factor 2 Relacionado con NF-E2/fisiología , Estrés Oxidativo , Western Blotting , Catalasa/metabolismo , Línea Celular , Citoprotección , Células Epiteliales/metabolismo , Citometría de Flujo , Técnica del Anticuerpo Fluorescente Indirecta , Glutatión/metabolismo , Humanos , Peróxido de Hidrógeno/toxicidad , Cristalino/metabolismo , Oxidantes/toxicidad , Fosforilación , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Superóxido Dismutasa/metabolismo , Transfección , Respuesta de Proteína Desplegada/fisiología , eIF-2 Quinasa/metabolismoRESUMEN
OBJECTIVES: This study aimed to investigate the protective role of heme oxygenase-1 (HO-1) in H2O2-induced oxidative stress and apoptosis in human lens epithelial cells (hLEC; SRA01/04). METHODS: SRA01/04 cells were exposed to a hydrogen peroxide (H2O2) concentration gradient and inducers of HO-1 such as cobalt protoporphyrin (CoPP) and zinc protoporphyrin (ZnPP), respectively. In addition, an RNA silencing experiment was conducted to investigate the HO-1 function in this study. A Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability. Western blot and ELISA were used to detect the level of HO-1 expression. In our study, hLECs were exposed to 400 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with 10µΜ CoPP or 10µΜ ZnPP, respectively. Double immunofluorescence staining was used for cell identification and the qualitative expression of HO-1. Expression of HO-1 was monitored using Western blot and ELISA. Intracellular reactive oxygen species (ROS) were detected by flow cytometry analyses; commercial enzymatic kits were used to measure the levels of glutathione (GSH), as well as superoxide dismutase (SOD). The proportion of cell apoptosis was quantified by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The expression of caspase family (-8, -3) proteins was measured by Western blot analysis. RESULTS: HO-1 significantly restored the cell viability under H2O2 injury via reducing the generation of ROS and increasing the levels of SOD and GSH activity. Moreover, HO-1 also inhibited H2O2-induced caspase-8 and caspase-3 proteins, thus significantly reducing the apoptosis of SRA01/04. An RNA silencing experiment demonstrated the increased resistance of LECs to oxidative stress specifically for increased levels of HO-1. CONCLUSIONS: These findings suggested that HO-1 protects human lens epithelial cells from H2O2-induced oxidant stress by upregulating antioxidant enzyme activity, reducing ROS generation, and thus inhibiting caspase family-dependent apoptosis.
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Apoptosis/efectos de los fármacos , Células Epiteliales/metabolismo , Hemo-Oxigenasa 1/fisiología , Peróxido de Hidrógeno/farmacología , Cristalino/metabolismo , Estrés Oxidativo/efectos de los fármacos , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Supervivencia Celular/efectos de los fármacos , Glutatión Sintasa/metabolismo , Humanos , Superóxido Dismutasa/metabolismoRESUMEN
PURPOSE: Fibrotic cataracts, including anterior subcapsular cataract (ASC) as well as posterior capsule opacification (PCO), are a common vision-threatening cause worldwide. Still, little is known about the underlying mechanisms. Here, we demonstrate a miRNA-based pathway regulating the pathological fibrosis process of lens epithelium. METHODS: Gain- and loss-of-function approaches, as well as multiple fibrosis models of the lens, were applied to validate the crucial role of two miR-1225 family members in the TGF-ß2 induced PCO model of human LECs and injury-induced ASC model in mice. RESULTS: Both miR-1225-3p and miR-1225-5p prominently stimulate the migration and EMT process of lens epithelial cells (LECs) in vitro as well as lens fibrosis in vivo. Moreover, we demonstrated that the underlying mechanism for these effects of miR-1225-5p is via directly targeting Keap1 to regulate Keap1/Nrf2 signaling. In addition, evidence showed that Keap1/Nrf2 signaling is activated in the TGF-ß2 induced PCO model of human LECs and injury-induced ASC model in mice, and inhibition of the Nrf2 pathway can significantly reverse the process of LECs EMT as well as lens fibrosis. CONCLUSIONS: These results suggest that blockade of miR-1225-5p prevents lens fibrosis via targeting Keap1 thereby inhibiting Nrf2 activation. The 'miR-1225-Keap1-Nrf2' signaling axis presumably holds therapeutic promise in the treatment of fibrotic cataracts.
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Catarata , Modelos Animales de Enfermedad , Fibrosis , Proteína 1 Asociada A ECH Tipo Kelch , Ratones Endogámicos C57BL , MicroARNs , Factor 2 Relacionado con NF-E2 , Transducción de Señal , Animales , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , MicroARNs/genética , Ratones , Proteína 1 Asociada A ECH Tipo Kelch/genética , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Humanos , Catarata/metabolismo , Catarata/genética , Catarata/patología , Cristalino/metabolismo , Cristalino/patología , Regulación de la Expresión Génica , Células Cultivadas , Células Epiteliales/metabolismo , Western Blotting , Movimiento Celular , Transición Epitelial-MesenquimalRESUMEN
AIM: To explore the effect of silent information regulator factor 2-related enzyme 1 (SIRT1) on modulating apoptosis of human lens epithelial cells (HLECs) and alleviating lens opacification of rats through suppressing endoplasmic reticulum (ER) stress. METHODS: HLECs (SRA01/04) were treated with varying concentrations of tunicamycin (TM) for 24h, and the expression of SIRT1 and C/EBP homologous protein (CHOP) was assessed using real-time quantitative polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence. Cell morphology and proliferation was evaluated using an inverted microscope and cell counting kit-8 (CCK-8) assay, respectively. In the SRA01/04 cell apoptosis model, which underwent siRNA transfection for SIRT1 knockdown and SRT1720 treatment for its activation, the expression levels of SIRT1, CHOP, glucose regulated protein 78 (GRP78), and activating transcription factor 4 (ATF4) were examined. The potential reversal of SIRT1 knockdown effects by 4-phenyl butyric acid (4-PBA; an ER stress inhibitor) was investigated. In vivo, age-related cataract (ARC) rat models were induced by sodium selenite injection, and the protective role of SIRT1, activated by SRT1720 intraperitoneal injections, was evaluated through morphology observation, hematoxylin and eosin (H&E) staining, Western blotting, and RT-PCR. RESULTS: SIRT1 expression was downregulated in TM-induced SRA01/04 cells. Besides, in SRA01/04 cells, both cell apoptosis and CHOP expression increased with the rising doses of TM. ER stress was stimulated by TM, as evidenced by the increased GRP78 and ATF4 in the SRA01/04 cell apoptosis model. Inhibition of SIRT1 by siRNA knockdown increased ER stress activation, whereas SRT1720 treatment had opposite results. 4-PBA partly reverse the adverse effect of SIRT1 knockdown on apoptosis. In vivo, SRT1720 attenuated the lens opacification and weakened the ER stress activation in ARC rat models. CONCLUSION: SIRT1 plays a protective role against TM-induced apoptosis in HLECs and slows the progression of cataract in rats by inhibiting ER stress. These findings suggest a novel strategy for cataract treatment focused on targeting ER stress, highlighting the therapeutic potential of SIRT1 modulation in ARC development.
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PURPOSE: To explore the correlation of endoplasmic reticulum stress (ERS) and oxidative stress (OS), and the protective effect of Sestrin2 (SESN2) on human lens epithelial cells (HLECs). METHODS: Tunicamycin (TM) was used to induce ERS in HLECs. 4-Phenylbutyric acid (4-PBA) was used to inhibit ERS. Eupatilin applied to HLECs as SESN2 agonist. SESN2 expression was knocked down via si-RNA in HLECs. The morphological changes of HLECs were observed by microscope. ER-tracker to evaluate ERS, ROS production assay to measure ROS, flow cytometry to calculate cell apoptosis rate. Immunofluorescence to observe Nrf2 translocation, and effects of TM or EUP on SESN2. Western blot and qPCR were used to evaluate the expression of GRP78, PERK, ATF4, CHOP, Nrf2, and SESN2 expression in HLECs with different treatment groups. RESULTS: ERS can elevate the expression of ROS and Nrf2 to induce OS. Upregulation of SESN2 was observed in ERS-mediate OS. Overexpression of SESN2 can reduce the overexpression of ERS-related protein GRP78, PERK, ATF4, proapoptotic protein CHOP, OS-related protein Nrf2, as well as ROS, and alleviate ERS injury at the same time. Whereas knockdown of SESN2 can upregulate the expression of GRP78, PERK, ATF4, CHOP, Nrf2, ROS, and deteriorate ERS damage. CONCLUSIONS: ERS can induce OS, they form a vicious cycle to induce apoptosis in HLECs, which may contribute to cataract formation. SESN2 could protect HLECs against the apoptosis by regulating the vicious cycle between ERS and OS.
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Apoptosis , Western Blotting , Catarata , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico , Células Epiteliales , Cristalino , Proteínas Nucleares , Estrés Oxidativo , Humanos , Estrés del Retículo Endoplásmico/fisiología , Cristalino/metabolismo , Cristalino/patología , Células Epiteliales/metabolismo , Catarata/metabolismo , Catarata/patología , Catarata/genética , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Células Cultivadas , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Citometría de Flujo , Factor 2 Relacionado con NF-E2/metabolismo , Factor 2 Relacionado con NF-E2/genética , Regulación de la Expresión Génica , SestrinasRESUMEN
BACKGROUND: Corneal edema (CE) affects the outcome of phacoemulsification. Effective ways to predict the CE after phacoemulsification are needed. METHODS: On the basis of data from patients conforming to the protocol of the AGSPC trial, 17 variables were selected to predict CE after phacoemulsification by constructing a CE nomogram through multivariate logistic regression, which was improved via variable selection with copula entropy. The prediction models were evaluated using predictive accuracy, the area under the receiver operating characteristic curve (AUC), and decision curve analysis (DCA). RESULTS: Data from 178 patients were used to construct prediction models. After copula entropy variable selection, which shifted the variables used for prediction in the CE nomogram from diabetes, best corrected visual acuity (BCVA), lens thickness and cumulative dissipated energy (CDE) to CDE and BCVA in the Copula nomogram, there was no significant change in predictive accuracy (0.9039 vs. 0.9098). There was also no significant difference in AUCs between the CE nomogram and the Copula nomogram (0.9637, 95% CI 0.9329-0.9946 vs. 0.9512, 95% CI 0.9075-0.9949; p = 0.2221). DCA suggested that the Copula nomogram has clinical application. CONCLUSIONS: This study obtained a nomogram with good performance to predict CE after phacoemulsification, and showed the improvement of copula entropy for nomogram models.
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The fluidics system is an indispensable and primary component of phacoemulsification. Both the gravity-fluidics system and active-fluidics system are commonly used in practice. The irrigation pressure of the gravity-fluidics system is determined by the bottle height, which is relatively constant, while the active-fluidics system is paired with a cassette that contains pressure sensors to monitor intraocular pressure changes. The active-fluidics system allows surgeons to preset a target intraocular pressure value, and it replenishes the fluids proactively; thus, the intraocular pressure is consistently maintained near the target value. Under such circumstances, stable intraocular pressure and anterior chamber volume values could be acquired. Research on surgical safety, efficiency and results have reported several strengths of the active-fluidics system. It is also advantageous in some complicated cataract surgeries. However, the system is not widely used at present, mainly due to its low penetration rate and high equipment cost. Some of its updates such as the new Active Sentry handpiece showed potential superiority in laboratory studies recently, but there is still further research to be conducted. This article gives an overview of the mechanism and performance of the active-fluidics system, and it is expected to provide clues for future research.
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PURPOSE: This study investigated the protective effect of carbon monoxide releasing molecule-3 (CORM-3), the classical donor of carbon monoxide, on selenite-induced cataract in rats and explore its possible mechanism. METHODS: Sprague-Dawley rat pups treated with sodium selenite (Na2SeO3) were chosen as the cataract model. Fifty rat pups were randomly divided into 5 groups: Control group, Na2SeO3 (3.46 mg/kg) group, low-dose CORM-3 (8 mg/kg/d) + Na2SeO3 group, high-dose CORM-3 (16 mg/kg/d) + Na2SeO3 group, and inactivated CORM-3 (iCORM-3) (8 mg/kg/d) + Na2SeO3 group. The protective effect of CORM-3 was tested by lens opacity scores, hematoxylin and eosin staining, TdT-mediated dUTP nick-end labeling assay, and enzyme-linked immunosorbent assay. Besides, quantitative real-time PCR and western blotting were used for mechanism validation. RESULTS: Na2SeO3 induced nuclear cataract rapidly and stably, and the achievement ratio of Na2SeO3 group was 100%. CORM-3 alleviated lens opacity of selenite-induced cataract and attenuated the morphological changes of the rat lens. The levels of antioxidant enzymes GSH and SOD in rat lens were also increased by CORM-3 treatment. CORM-3 significantly reduced the ratio of apoptotic lens epithelial cells, besides, CORM-3 decreased the expression of Cleaved Caspase-3 and Bax induced by selenite and increased the expression of Bcl-2 in rat lens inhibited by selenite. Moreover, Nrf-2 and HO-1 were upregulated and Keap1 was downregulated after CORM-3 treatment. While iCORM-3 did not exert the same effect as CORM-3. CONCLUSIONS: Exogenous CO released from CORM-3 alleviates oxidative stress and apoptosis in selenite-induced rat cataract via activating Nrf2/HO-1 pathway. CORM-3 may serve as a promising preventive and therapeutic strategy for cataract.
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Catarata , Ácido Selenioso , Ratas , Animales , Ratas Sprague-Dawley , Ácido Selenioso/toxicidad , Monóxido de Carbono/efectos adversos , Monóxido de Carbono/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Catarata/inducido químicamente , Catarata/prevención & control , Catarata/tratamiento farmacológico , Estrés Oxidativo , ApoptosisRESUMEN
INTRODUCTION: Posterior capsular opacification (PCO) is the most common complication of cataract surgery. In this study, we develop a model to quantitatively predict the probability of Nd:YAG laser capsulotomy for vision-threatening PCO to improve the life quality of postoperative patients. METHODS: A registry analysis of cataract procedures performed between the years 2010 and 2021. Following the screening of 16,802 patients (25,883 eyes), 9768 patients (eyes) were enrolled. The cohort was randomly divided into two groups: training (n = 6838) and validation (n = 2930). To identify relevant risk factors, univariate, multivariate, and Least Absolute Shrinkage and Selection Operator (LASSO) algorithm Cox regression analysis were employed, and a nomogram was created to demonstrate the prediction result. RESULTS: At 5 years, the overall cumulative incidence of Nd:YAG laser capsulotomy was 12.0% (1169/9768). The following variables were included in the prediction model: sex [hazard ratio (HR) = 1.53, 95% CI 1.32-1.76], age (HR = 0.71, 95% CI 0.56-0.88), intraocular lens (IOL) material (HR = 2.65, 95% CI 2.17-3.24), high myopia (HR = 2.28, 95% CI 1.90-2.75), and fibrinogen (HR = 0.79, 95% CI 0.72-0.88). In the validation cohort, the area under the curve (AUC) of 1-, 3-, and 5-year predictions for Nd:YAG laser capsulotomy were 0.702, 0.691, and 0.688, respectively. For a subgroup of patients with high myopia, the protective effect of hydrophobic IOL disappeared (HR = 0.68, 95% CI 0.51-1.12, P = 0.127). CONCLUSION: This model could predict the probability of Nd:YAG laser capsulotomy for vision-threatening PCO after cataract surgery by taking into account factors such as age, gender, IOL material, high myopia, and fibrinogen. Meanwhile, implantation of a hydrophobic IOL in individuals with high myopia did not demonstrate a protective impact against vision-threatening PCO.
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Background: Due to media opacity, it is usually difficult to accurately evaluate the postoperative visual acuity in cataracts patients. As a small and portable tool, the critical flicker fusion frequency (CFF) device reflects the temporal resolution of visual function and has been widely used in clinical research. However, poor understanding of the technique and equipment limitations have restricted its clinical application in China. Main text: There was a decrease in the CFF value in various ophthalmic diseases, indicating that the CFF is sensitive to detect visual functional changes. A number of studies have shown that the CFF test can accurately distinguish patients with simple cataracts from those with cataracts combined with fundus disease, and, as a visual test, it can more accurately predict postoperative visual acuity without being affected by media opacity. This study comprehensive reviews the basic principles of CFF and its application in ophthalmology, especially in cataracts. Conclusions: As one of the tools for dynamic visual function detection, the CFF test could help doctors to assess the possible presence of fundus disease in cataracts patients, especially in eyes with dense cataracts, and more precisely provide a reasonable visual prognosis than other available visual tests.
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PURPOSE: This study aimed to investigate the protective effects of piceatannol (PIC) on selenite-induced cataracts in Sprague-Dawley rats and explore its therapeutic effects as an antioxidant. METHODS: Thirty-two eight-day-old rat pups were randomly divided into four groups, with eight pups in each of them. Group 1, as the control group, was injected with the same amount of saline, while Groups 2-4 were administered with sodium selenite (3.46 mg/kg) subcutaneously into the neck on postpartum day 10 for cataract induction. Without further treatment, Group 2 served as the control model, while Groups 3 and 4 (low- and high-dose PIC-treated) had intraperitoneal piceatannol from day 8 to day 17 at doses of 10 mg/kg and 20 mg/kg, respectively. On postpartum day 17, after the last injection, the rat pups were examined for cataract grade by slit lamp, and the lenses of every group were isolated for oxidative damage indicators and further analysis. SRA01/04 cells were exposed to 600 µM H2O2 for 24 hours with or without pretreatment with 10µÐ piceatannol. Cell viability was tested by CCK-8 assay and cell apoptosis was evaluated by AnnexinV-PE/7AAD assay. RESULTS: This study determined that compared with the model group, the degree of lens opacity was significantly reduced in PIC-treated groups. The histopathological damage of the lenses in the PIC-treated groups improved compared to the model group. There were fewer signs of lesions, such as vacuoles and atrophy. The biochemical results indicated that malondialdehyde (MDA) content of the PIC-treated groups were downregulated and the antioxidant enzyme activities (GSH and catalase) and antioxidant status (SOD) were upregulated compared with the model group. In vitro, piceatannol significantly restored cell viability and cell apoptosis under H2O2 injury. CONCLUSION: Pretreatment with piceatannol may achieve a protective effect on cataract development through upregulating antioxidant enzyme activity.
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Catarata , Estrés Oxidativo , Estilbenos , Animales , Antioxidantes/farmacología , Catarata/inducido químicamente , Catarata/prevención & control , Femenino , Glutatión/metabolismo , Peróxido de Hidrógeno/toxicidad , Cristalino/patología , Malondialdehído , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Selenito de Sodio/toxicidad , Estilbenos/farmacologíaRESUMEN
Oxidation is an essential factor during cataract development. Autophagy, usually a cytoprotective process, is always found elevated in lens epithelial cells under oxidation, yet its roles and associated molecular mechanisms under such circumstances are rarely elucidated. Herein, we extracted and re-analyzed the RNA sequencing data of the GSE161701 dataset from the Gene Expression Omnibus database to identify the differentially expressed mRNAs and lncRNAs by using the R package "DESeq2". Further analyses of gene ontology and KEGG enrichment were implemented via the packages "clusterProfiler" and "enrichplot". We found that after the knockout of ATG7, differentially expressed genes were more associated with hemopoiesis, vasculature development, axonogenesis, and hypoxia regulation. When stimulated with H2O2, LECs displayed a gene expression profile correlating with apoptotic and proliferative pathways, such as the MAPK signaling pathway and FoxO signaling pathway. The differentially expressed gene profiles of the two types of LECs (wild type and ATG7 deficient) under oxidation were distinct to a large extent. Furthermore, 1,341 up-regulated and 1912 down-regulated differential mRNAs and 263 up-regulated and 336 down-regulated differential lncRNAs between these two types of LECs subjected to H2O2 were detected, among which 292 mRNAs and 24 lncRNAs possibly interacted with ten cataract-related miRNAs. A competing endogenous lncRNA-miRNA-mRNA network based on such interactions was finally constructed.
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INTRODUCTION: The active-fluidics system is a new irrigation system of phacoemulsification that automatically detects and maintains stable intraocular pressure at the set value. This trial is designed to compare the efficacy, visual outcomes, safety and patients' subjective perceptions of cataract surgery with the active-fluidics system and gravity-fluidics system. METHODS AND ANALYSIS: This trial will recruit 110 patients with age-related cataract at the Chinese People's Liberation Army (PLA) General Hospital (Beijing, China) and they will be randomly assigned to the active-fluidics group and gravity-fluidics group in a ratio of 1:1 to have phacoemulsification. Patients will be followed up at 1 day, 1 week, 1 month and 3 months postoperatively. The primary outcomes are the cumulative dissipated energy and best corrected visual acuity. Secondary outcomes include: estimated fluid usage, U/S time, total aspiration time, intraocular pressure, corneal endothelium parameters, retinal thickness, macular superficial vessel density, pain scores, scores of the Cataract surgery Patient-Reported Outcome Measures Questionnaire and the complication rates. The data will be independently analysed by the statistical team, who will be masked for the allocation information as participants are. ETHICS AND DISSEMINATION: This study was approved by the Ethics Committee of Chinese PLA General Hospital (approval no. S2021-068-01). Informed consent will be obtained from each participant. All the results will be published in peer-reviewed journals and used for scholarly communication or technical guidance. Protocol version 1.0. TRIAL REGISTRATION NUMBER: Chinese Clinical Trial Registry (ChiCTR2100044409).
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Extracción de Catarata , Catarata , Facoemulsificación , Extracción de Catarata/métodos , Humanos , Facoemulsificación/métodos , Estudios Prospectivos , Ensayos Clínicos Controlados Aleatorios como Asunto , Agudeza VisualRESUMEN
BACKGROUND: To figure out the efficacy, effects, safety and patient's subjective perceptions of phacoemulsification with the active-fluidics system (AFS). PATIENTS AND METHODS: This was a prospective, randomized, double-masked, controlled clinical study. Age-related cataract patients were recruited and randomly assigned to the AFS group and gravity-fluidics system (GFS) group in a ratio of 1:1 to have phacoemulsification. Participants were followed up at one day, one week, one month and three months postoperatively (Chinese Clinical Trial Registry, ChiCTR2100044409). RESULTS: The overall included participants were 107 finally. The total aspiration time of the AFS group was significantly less than that of the GFS group (p = .020), while no significant difference existed in cumulative dissipated energy and estimated fluid usage between the two groups. The best corrected visual acuity was significantly better in the AFS group at one day and one week postoperatively (p = .002, p = .038 respectively). The recovery of central corneal thickening and macular superficial vasculature increase was earlier in the AFS group. The central retinal thickness was significantly higher in the GFS group at one month and three months postoperatively (p = .029, p = .016 respectively). The incidence of corneal adverse events was higher in GFS group (p = .035). No serious adverse events occurred in either group. Pain scores and the scores of Cat-PROM5 questionnaire of the AFS group were significantly lower than that of the GFS group (p = .011, p = .002 respectively). CONCLUSION: AFS improves the efficiency, effects, safety and patients' subjective perceptions of phacoemulsification compared with GFS. It is worthwhile to promote its application in cataract surgery.KEY MESSAGESThe active-fluidics system automatically detects and maintains stable intraocular pressure at the set value.The active-fluidics system improves the efficiency, effects, safety and patients' subjective perceptions in phacoemulsification.