ABSTRACT
Application of stable isotopically labelled (SIL) molecules in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI) over a series of time points allows the temporal and spatial dynamics of biochemical reactions to be tracked in a biological system. However, these large kinetic MSI datasets and the inherent variability of biological replicates presents significant challenges to the rapid analysis of the data. In addition, manual annotation of downstream SIL metabolites involves human input to carefully analyse the data based on prior knowledge and personal expertise. To overcome these challenges to the analysis of spatiotemporal MALDI-MSI data and improve the efficiency of SIL metabolite identification, a bioinformatics pipeline has been developed and demonstrated by analysing normal bovine lens glucose metabolism as a model system. The pipeline consists of spatial alignment to mitigate the impact of sample variability and ensure spatial comparability of the temporal data, dimensionality reduction to rapidly map regional metabolic distinctions within the tissue, and metabolite annotation coupled with pathway enrichment modules to summarise and display the metabolic pathways induced by the treatment. This pipeline will be valuable for the spatial metabolomics community to analyse kinetic MALDI-MSI datasets, enabling rapid characterisation of spatio-temporal metabolic patterns from tissues of interest.
Subject(s)
Glucose , Lens, Crystalline , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Animals , Cattle , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Lens, Crystalline/metabolism , Glucose/metabolism , Isotope Labeling/methods , Workflow , Metabolomics/methods , Data Analysis , Metabolic Networks and PathwaysSubject(s)
Drainage , Fluorocarbons , Retinal Detachment , Retinal Telangiectasis , Humans , Retinal Detachment/diagnosis , Retinal Detachment/surgery , Retinal Detachment/etiology , Fluorocarbons/administration & dosage , Drainage/methods , Retinal Telangiectasis/diagnosis , Retinal Telangiectasis/complications , Male , Lens, Crystalline , Exudates and Transudates , Visual Acuity , Endotamponade/methodsABSTRACT
PURPOSE: The epithelial-mesenchymal transition of human lens epithelial cells plays a role in posterior capsule opacification, a fibrotic process that leads to a common type of cataract. Hyaluronic acid has been implicated in this fibrosis. Studies have investigated the role of transforming growth factor (TGF)-ß2 in epithelial-mesenchymal transition. However, the role of TGF-ß2 in hyaluronic acid-mediated fibrosis of lens epithelial cell remains unknown. We here examined the role of TGF-ß2 in the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells. METHODS: Cultured human lens epithelial cells (HLEB3) were infected with CD44-siRNA by using the Lipofectamine 3000 transfection reagent. The CCK-8 kit was used to measure cell viability, and the scratch assay was used to determine cell migration. Cell oxidative stress was analyzed in a dichloro-dihydro-fluorescein diacetate assay and by using a flow cytometer. The TGF-ß2 level in HLEB3 cells was examined through immunohistochemical staining. The TGF-ß2 protein level was determined through western blotting. mRNA expression levels were determined through quantitative real-time polymerase chain reaction. RESULTS: Treatment with hyaluronic acid (1.0 µM, 24 h) increased the epithelial-mesenchymal transition of HLEB3 cells. The increase in TGF-ß2 levels corresponded to an increase in CD44 levels in the culture medium. However, blocking the CD44 function significantly reduced the TGF-ß2-mediated epithelial-mesenchymal transition response of HLEB3 cells. CONCLUSIONS: Our study showed that both CD44 and TGF-ß2 are critical contributors to the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells, and that TGF-ß2 in epithelial-mesenchymal transition is regulated by CD44. These results suggest that CD44 could be used as a target for preventing hyaluronic acid-induced posterior capsule opacification. Our findings suggest that CD44/TGF-ß2 is crucial for the hyaluronic acid-induced epithelial-mesenchymal transition of lens epithelial cells.
Subject(s)
Cell Movement , Epithelial Cells , Epithelial-Mesenchymal Transition , Hyaluronan Receptors , Hyaluronic Acid , Lens, Crystalline , Transforming Growth Factor beta2 , Humans , Epithelial-Mesenchymal Transition/drug effects , Epithelial-Mesenchymal Transition/physiology , Hyaluronic Acid/pharmacology , Hyaluronan Receptors/metabolism , Transforming Growth Factor beta2/pharmacology , Transforming Growth Factor beta2/metabolism , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Lens, Crystalline/cytology , Lens, Crystalline/drug effects , Lens, Crystalline/metabolism , Cell Movement/drug effects , Cell Survival/drug effects , Oxidative Stress/drug effects , Oxidative Stress/physiology , Blotting, Western , Capsule Opacification/metabolism , Capsule Opacification/pathology , Real-Time Polymerase Chain Reaction , Flow Cytometry , Immunohistochemistry , Cells, CulturedSubject(s)
Magnetic Resonance Imaging , Humans , Male , Lens, Crystalline/diagnostic imaging , FemaleABSTRACT
The aging eye undergoes the same progressive crosslinking which occurs throughout the body, resulting in increased rigidity of ocular connective tissues including the lens and the sclera which impact ocular functions. This offers the potential for a scleral treatment that is based on restoring normal biomechanical movements. Laser Scleral Microporation is a laser therapy that evaporates fractional areas of crosslinked tissues in the sclera, reducing ocular rigidity over critical anatomical zones of the accommodation apparatus, restoring the natural dynamic range of focus of the eye. Although controversial and challenged, an alternative theory for presbyopia is Schachar's theory that suggests a reduction in the space between the ciliary processes and the crystalline lens. Widening of this space with expansion bands has been shown to aid near vision in people with presbyopia, a technique that has been used in the past but seems to be obsolete now. The use of drugs has been used in the treatment of presbyopia, either to cause pupil miosis to increase depth of focus, or an alteration in refractive error (to induce myopia in one eye to create monovision). Drugs and laser ablation of the crystalline lens have been used with the aim of softening the hardened lens. Poor nutrition and excess exposure to ultraviolet light have been implicated in the onset of presbyopia. Dietary nutritional supplements, lifestyle changes have also been shown to improve accommodation and the question arises whether these could be harnessed in a treatment for presbyopia as well.
Subject(s)
Presbyopia , Sclera , Presbyopia/therapy , Presbyopia/physiopathology , Humans , Lens, Crystalline , Laser Therapy/methods , Accommodation, Ocular/physiologyABSTRACT
Purpose: The role of specific extracellular matrix (ECM) molecules in lens cell development and regeneration is poorly understood, as appropriate cellular models are lacking. Here, a laminin-based lens cell in vitro induction system was developed to study the role of laminin in human lens epithelial stem/progenitor cell (LES/PC) development. Methods: The human embryonic stem cell-based lens induction system followed a three-stage protocol. The expression profile of laminins during lens induction was screened, and laminin-511 (LN511) was tested as a candidate substitute. LN511 induction system cellular and molecular features, including induction efficiency, transcription factor expression related to different lens development stages, ECM alterations, and Hippo/YAP signaling, were evaluated. Results: LAMA5, LAMB1, and LAMC1 were highly expressed around the time of LES/PC derivation. We chose LN511 (product of LAMA5, LAMB1, and LAMC1) and found that it considerably enhanced lens cell induction efficiency, compared to that in Matrigel-coated culture, as more and larger lentoid bodies were detected. Notably, LES/PC induction efficiency improved by promoting lens specification-related transcription factor expression and cell proliferation. Transcriptome analysis revealed that compared to those with Matrigel, ECM accumulation and cell adhesion were downregulated in the LN511 system. Hippo/YAP signaling was hypoactive during LES/P-like cell generation, and small molecule inhibitors of YAP/TAZ activity upregulated LES/PC marker expression and promoted the efficiency of LES/P-like cell derivation. Conclusions: The laminin isoform LN511 is a reliable substitute for the LES/P-like cell induction system, and LN511-YAP acted as efficient modulators of LES/PC derivation; this contributes to knowledge of the role of the ECM in human lens development.
Subject(s)
Cell Differentiation , Cell Proliferation , Epithelial Cells , Laminin , Lens, Crystalline , Humans , Laminin/metabolism , Lens, Crystalline/cytology , Lens, Crystalline/metabolism , Epithelial Cells/metabolism , Epithelial Cells/drug effects , Cells, Cultured , Signal Transduction , Stem Cells/metabolism , Stem Cells/cytology , Transcription Factors/metabolism , Transcription Factors/genetics , Extracellular Matrix/metabolismABSTRACT
BACKGROUND: To assess the relationship between postoperative implantable collamer lens (ICL) vault and lens height obtained from two different measurements. METHODS: A retrospective case series study enrolled eyes with horizontally implanted ICL. Crystal lens rise (CLR) and the distance between STS plane and anterior crystalline lens surface (STSL) were measured in the horizontal and vertical directions using ultrasound biomicroscopy (UBM). We compared the differences in the parameters measured in both horizontal and vertical directions. The participants were categorized into three groups according to ciliary sulcus width (CSW) which is defined as the distance between the posterior angle of the iris and the anterior angle of the ciliary process: narrow CSW group (NSG); medium CSW group (MSG); and wide CSW group (WSG). The correlations between CLR/STSL and vault were examined in each of the three groups. Biased correlation analysis was used further to contrast the correlation between CLR/STSL and vault. RESULTS: This retrospective study included 223 myopic eyes. Vertical STSL (VSTSL) and vertical CLR (VCLR) exhibited significantly greater values compared to their horizontal counterparts (both P < 0.05). None of the indicators were statistically different between the three groups. In both NSG and MSG, STSL/CLR correlated with vault, while in WSG, only STSL correlated with vault (r=-0.316, P = 0.013). In contrast to HCLR, the correlation between HSTSL and vault remained after controlling for HCLR (r=-0.162, P = 0.015). CONCLUSIONS: STSL should deserve more attention in the preoperative evaluation of ICL compared to CLR especially when CSW is large.
Subject(s)
Ciliary Body , Lens Implantation, Intraocular , Microscopy, Acoustic , Myopia , Phakic Intraocular Lenses , Humans , Retrospective Studies , Male , Female , Ciliary Body/diagnostic imaging , Ciliary Body/pathology , Pilot Projects , Adult , Myopia/surgery , Myopia/physiopathology , Lens, Crystalline/diagnostic imaging , Young Adult , Middle Aged , Visual Acuity/physiology , Refraction, Ocular/physiologyABSTRACT
Cataracts are the world's number one blinding eye disease. Cataracts can only be effectively treated surgically, although there is a chance of surgical complications. One of the pathogenic processes of cataracts is oxidative stress, which closely correlated with pyroptosis. SIRT1 is essential for the regulation of pyroptosis. Nevertheless, the role of SIRT1 in formation of cataracts is unclear. In this work, we developed an in vitro model of shortwave blue light (SWBL)-induced scotomization in human lens epithelial cells (HLECs) and an in vivo model of SWBL-induced cataracts in rats. The study aimed to understand how the SIRT1/NF-κB/NLRP3 pathway functions. Additionally, the evaluation included cell death and the release of lactate dehydrogenase (LDH), a cytotoxicity marker, from injured cells. First, we discovered that SWBL exposure resulted in lens clouding in Sprague- Dawley (SD) rats and that the degree of clouding was positively linked to the duration of irradiation. Second, we discovered that SIRT1 exhibited antioxidant properties and was connected to the NF-κB/NLRP3 pathway. SWBL irradiation inhibited SIRT1 expression, exacerbated oxidative stress, and promoted nuclear translocation of NF-κB and the activation of the NLRP3 inflammasome, which caused LEC pyroptosis and ultimately led to cataract formation. Transient transfection to increase the expression of SIRT1 decreased the protein expression levels of NF-κB, NLRP3, caspase-1, and GSDMD, inhibited HLEC pyroptosis, and reduced the release of LDH, providing a potential method for cataract prevention and treatment.
Subject(s)
Cataract , Epithelial Cells , Lens, Crystalline , NF-kappa B , NLR Family, Pyrin Domain-Containing 3 Protein , Pyroptosis , Sirtuin 1 , Animals , Humans , Rats , Blotting, Western , Blue Light/adverse effects , Cataract/metabolism , Cataract/pathology , Cataract/etiology , Cells, Cultured , Disease Models, Animal , Epithelial Cells/metabolism , Epithelial Cells/radiation effects , Lens, Crystalline/radiation effects , Lens, Crystalline/metabolism , NF-kappa B/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Oxidative Stress , Pyroptosis/physiology , Pyroptosis/radiation effects , Rats, Sprague-Dawley , Signal Transduction/physiology , Sirtuin 1/metabolismABSTRACT
PURPOSE: To evaluate the impact of anterior chamber phakic intraocular lens (pIOL) on swept-source optical coherence tomography (SS-OCT) biometric measurements and IOL power calculation. METHODS: This retrospective analysis of 67 eyes of 49 patients with previous anterior chamber pIOL implantation analyzed the accuracy of automatic segmentation of the anterior surface of the crystalline lens and its impact on anterior chamber depth (ACD, measured from epithelium to lens), lens thickness measurements, and IOL power calculation. The sample was divided into two groups: correct detection of the anterior surface of the crystalline lens and inaccurate detection. Segmentation of eyes from the inaccurate detection group was manually corrected and ACD and lens thickness were calculated using ImageJ software. IOL power was calculated using 7 formulas for both measurements. RESULTS: The anterior surface of the crystalline lens was mis-identified in 13 (19.4%) eyes. ACD was underestimated (Δ -0.85 ± 0.33 mm, P < .001) and lens thickness was overestimated (Δ +0.81 ± 0.25 mm, P < .001). Manual correction changed the target spherical equivalent only in the Haigis formula (P = .009). After correction for segmentation bias, the Pearl DGS, Cooke K6, and EVO 2.0 formulas showed the lowest prediction error, with the Pearl DGS showing greatest accuracy within ±1.00 diopters of prediction error range (81.0%). CONCLUSIONS: SS-OCT biometry misidentifies the anterior surface of the crystalline lens in a significant proportion, resulting in significant IOL power calculation error in the Haigis formula. Manual proofing of segmentation is mandatory in every patient with anterior chamber pIOL implantation. After correct segmentation, the Pearl DGS, Cooke K6, and EVO seem to be the best formulas. [J Refract Surg. 2024;40(8):e562-e568.].
Subject(s)
Anterior Chamber , Biometry , Lens Implantation, Intraocular , Phakic Intraocular Lenses , Tomography, Optical Coherence , Humans , Tomography, Optical Coherence/methods , Biometry/methods , Retrospective Studies , Male , Adult , Female , Anterior Chamber/diagnostic imaging , Refraction, Ocular/physiology , Middle Aged , Optics and Photonics , Lens, Crystalline/diagnostic imaging , Myopia/surgery , Myopia/physiopathology , Visual Acuity/physiology , Young AdultABSTRACT
Antioxidant therapies are of interest in the prevention and management of ocular disorders such as cataracts. Although an active area of interest, topical therapy with antioxidants for the treatment of cataracts is complicated by multiple ocular anatomical barriers, product stability, and solubility. Entrapment and delivery of antioxidants with poly(lactic-co-glycolic acid) nanoparticles is a possible solution to these challenges, however, little is known regarding their effects in vitro or in vivo. Our first aim was to investigate the impact of blank and lutein loaded PLGA nanoparticles on viability and development of reactive oxygen species in lens epithelial cells in vitro. Photo-oxidative stress was induced by ultraviolet light exposure with cell viability and reactive oxygen species monitored. Next, an in vivo, selenite model was utilized to induce cataract formation in rodents. Eyes were treated topically with both free lutein and lutein loaded nanoparticles (LNP) at varying concentrations. Eyes were monitored for the development of anterior segment changes and cataract formation. The ability of nanodelivered lutein to reach the anterior segment of the eye was evaluated by liquid chromatography coupled to mass spectrometry of aqueous humor samples and liquid chromatography coupled to tandem mass spectrometry (targeted LC-MS/MS) of lenses. LNP had a minimal impact on the viability of lens epithelial cells during the short exposure timeframe (24 h) and at concentrations < 0.2 µg LNP/µl. A significant reduction in the development of reactive oxygen species was also noted. Animals treated with LNPs at an equivalent lutein concentration of 1,278 µg /mL showed the greatest reduction in cataract scores. Lutein delivery to the anterior segment was confirmed through evaluation of aqueous humor and lens sample evaluation. Topical treatment was not associated with the development of secondary keratitis or anterior uveitis when applied once daily for one week. LNPs may be an effective in the treatment of cataracts.
Subject(s)
Administration, Topical , Cataract , Lutein , Nanoparticles , Polylactic Acid-Polyglycolic Acid Copolymer , Animals , Lutein/pharmacology , Lutein/administration & dosage , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Nanoparticles/chemistry , Cataract/drug therapy , Rats , Lens, Crystalline/drug effects , Lens, Crystalline/metabolism , Reactive Oxygen Species/metabolism , Oxidative Stress/drug effects , Cell Survival/drug effects , Antioxidants/pharmacology , Antioxidants/administration & dosage , Humans , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Aqueous Humor/drug effects , Aqueous Humor/metabolism , Male , Cell Line , Lactic Acid/chemistry , Polyglycolic Acid/chemistryABSTRACT
In India, the prevailing approach to eye lens dosimetry is the placement of an existing dosemeter on the forehead region after slight modification serves as a dedicated Eye Lens Dosemeter. A methodology for estimating the eye lens dose in terms of the Hp(3) has been previously explored employing an algorithm based on the response characteristics of this dosemeter using ISO slab phantom. It was observed that the performance of the dosemeter in terms of Hp(3) using previous algorithm showed under response at higher angles of incidence and photon beams of energy < 200 keV. Further, study was conducted to modify the algorithm following the latest ISO recommendations. This involved generation of data from the response of existing dosemeter on a cylindrical phantom. The results of this study revealed better performance of the newly established algorithm in estimating eye lens dose in terms of Hp(3) when compared to the earlier algorithm.
Subject(s)
Algorithms , Lens, Crystalline , Phantoms, Imaging , Thermoluminescent Dosimetry , Humans , Calibration , Lens, Crystalline/radiation effects , Thermoluminescent Dosimetry/instrumentation , Thermoluminescent Dosimetry/methods , Radiation Dosage , Photons , Head/radiation effects , India , Radiation Protection/standards , Radiation Protection/instrumentationABSTRACT
PURPOSE: Autostereoscopic displays have become increasingly common, but their impact on ocular dimensions remains unknown. We sought to identify changes in the crystalline lens dimensions induced by autostereoscopic three-dimensional (3D) viewing. METHODS: Forty young adults (age: 22.6 ± 2.0 years, male/female: 15/25) were consecutively enrolled and randomly divided into two groups (3D and two-dimensional [2D] viewing groups) to watch a 30-min movie clip displayed in 3D or 2D mode on a tablet computer. The lens thickness (LT), diameter, curvature, decentration and tilt were measured with anterior segment optical coherence tomography under both non-accommodating (static) and accommodating conditions. RESULTS: In the static condition, the LT decreased by 0.03 ± 0.03 mm (p < 0.001) and the anterior radius of curvature (ARC) increased by 0.49 ± 0.59 mm (p = 0.001) post-3D viewing. In contrast, following 2D viewing, the ARC decreased by 0.23 ± 0.25 mm (p = 0.001). Additionally, the increase in the steep ARC post-3D viewing was greater in high-myopic eyes than low to moderate myopic eyes (p = 0.04). When comparing the accommodative with the static (non-accommodative) condition, for 3D viewing the lens decentration decreased (-0.03 ± 0.05 mm, p = 0.02); while for 2D viewing, the posterior curvature radius (-0.14 ± 0.20 mm, p = 0.006) and diameter (-0.13 ± 0.20 mm, p = 0.01) decreased. CONCLUSIONS: Viewing with the autostereoscopic 3D tablet could temporally decrease the thickness and curvature of the lens under non-accommodating conditions. However, its long-term effect requires further exploration.
Subject(s)
Accommodation, Ocular , Imaging, Three-Dimensional , Lens, Crystalline , Myopia , Tomography, Optical Coherence , Humans , Female , Male , Young Adult , Tomography, Optical Coherence/methods , Imaging, Three-Dimensional/methods , Lens, Crystalline/diagnostic imaging , Accommodation, Ocular/physiology , Myopia/physiopathology , Myopia/diagnosis , Refraction, Ocular/physiology , AdultABSTRACT
This study aims to investigate the hypothesis that Yes-associated protein (YAP) significantly regulates antioxidant potential and anti-apoptosis in UVB-induced cataract by exploring the underlying molecular mechanisms. To investigate the association between YAP and cataract, various experimental techniques were employed, including cell viability assessment, Annexin V FITC/PI assay, measurement of ROS production, RT-PCR, Western blot assay, and Immunoprecipitation. UVB exposure on human lens epithelium cells (HLECs) reduced total and nuclear YAP protein expression, increased cleaved/pro-caspase 3 ratios, decreased cell viability, and elevated ROS levels compared to controls. Similar Western blot results were observed in in vivo experiments involving UVB-treated mice. YAP knockdown in vitro demonstrated a decrease in the protein expression of FOXM1, Nrf2, and HO-1, which correlated with the mRNA expression, accompanied by an increase in cell apoptosis, caspase 3 activation, and the release of ROS. Conversely, YAP overexpression mitigated these effects induced by UVB irradiation. Immunoprecipitation revealed a FOXM1-YAP interaction. Notably, inhibiting FOXM1 decreased Nrf2 and HO-1, activating caspase 3. Additionally, administering the ROS inhibitor N-acetyl-L-cysteine (NAC) effectively mitigated the apoptotic effects induced by oxidative stress from UVB irradiation, rescuing the protein expression levels of YAP, FOXM1, Nrf2, and HO-1. The initial findings of our study demonstrate the existence of a feedback loop involving YAP, FOXM1, Nrf2, and ROS that significantly influences the cell apoptosis in HLECs under UVB-induced oxidative stress.
Subject(s)
Apoptosis , Cataract , Forkhead Box Protein M1 , NF-E2-Related Factor 2 , Oxidative Stress , Ultraviolet Rays , YAP-Signaling Proteins , Apoptosis/radiation effects , NF-E2-Related Factor 2/metabolism , NF-E2-Related Factor 2/genetics , Ultraviolet Rays/adverse effects , Humans , Animals , Forkhead Box Protein M1/metabolism , Forkhead Box Protein M1/genetics , Mice , Cataract/etiology , Cataract/metabolism , Cataract/pathology , YAP-Signaling Proteins/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Adaptor Proteins, Signal Transducing/genetics , Lens, Crystalline/metabolism , Lens, Crystalline/radiation effects , Transcription Factors/metabolism , Transcription Factors/genetics , Reactive Oxygen Species/metabolism , Male , Signal Transduction , Mice, Inbred C57BLABSTRACT
OBJECTIVES: Age-related cataract is the most common type of adult cataract and a leading cause of blindness. Currently, there are few reports on the establishment of animal models for age-related cataract. During the experimental breeding of Microtus fortis (M. fortis), we first observed that M. fortis aged 12 to 15 months could naturally develop cataracts. This study aims to explore the possibility of developing them as an animal model for age-related cataract via identifing and analyzing spontaneous cataract in M. fortis. METHODS: The 12-month-old healthy M. fortis were served as a control group and 12-month-old cataractous M. fortis were served as an experimental group. The lens transparency was observed using the slit-lamp biomicroscope. Hematoxylin and eosin staining was used to detect pathological changes in the lens. Biochemical detection methods were applied to detect blood routine, blood glucose levels, the serum activities of superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in both groups. Finally, real-time RT-PCR was used to detect the transcription levels of cataract-related genes in the lens of 2 groups. RESULTS: Compared with the control group, the lens of cataract M. fortis showed severely visible opacity, the structure of lens was destroyed seriously, and some pathological damage, such as swelling, degeneration/necrosis, calcification, hyperplasia, and fiber liquefaction were found in lens epithelial cells (LECs). The fibrous structure was disorganized and irregularly distributed with morgagnian globules (MGs) aggregated in the degenerated lens fibers. There was no statistically significant difference in blood glucose levels between the experimental and control groups (P>0.05). However, white blood cell (WBC) count (P<0.05), lymphocyte count (P<0.01), and lymphocyte ratio (P<0.05) were significantly decreased, while neutrophil percentage (P<0.05) and monocyte ratio (P<0.01) were significantly increased. The serum activities of SOD and GSH-Px (both P<0.05) were both reduced. The mRNAs of cataract-related genes, including CRYAA, CRYBA1, CRYBB3, Bsfp1, GJA3, CRYBA2, MIP, HspB1, DNase2B, and GJA8, were significantly downregultaed in the lenses of the experimental group (all P<0.05). CONCLUSIONS: There are significant differences in lens pathological changes, peroxidase levels, and cataract-related gene expression between cataract and healthy M. fortis. The developed cataract spontaneously in M. fortis is closely related to age, the cataract M. fortis might be an ideal animal model for the research of age-related cataract.
Subject(s)
Arvicolinae , Cataract , Glutathione Peroxidase , Lens, Crystalline , Superoxide Dismutase , Animals , Cataract/genetics , Cataract/pathology , Cataract/etiology , Lens, Crystalline/pathology , Glutathione Peroxidase/genetics , Glutathione Peroxidase/metabolism , Glutathione Peroxidase/blood , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolism , Aging , Disease Models, AnimalABSTRACT
BACKGROUND: Diabetic cataract (DC) is a common complication of diabetes and its etiology and progression are multi-factorial. In this study, the roles of specific protein 1 (SP1) and fibroblast growth factor 7 (FGF7) in DC development were explored. METHODS: DC cell model was established by treating SRA01/04 cells with high glucose (HG). MTT assay was conducted to evaluate cell viability. Transwell assay and wound-healing assay were performed to assess cell migration and invasion. Western blot assay and qRT-PCR assay were conducted to measure the expression of N-cadherin, E-cadherin, Collagen I, Fibronectin, SP1 and FGF7 expression. CHIP assay and dual-luciferase reporter assay were conducted to analyze the combination between FGF7 and SP1. RESULTS: FGF7 was upregulated in DC patients and HG-induced SRA01/04 cells. HG treatment promoted SRA01/04 cell viability, migration, invasion and epithelial-mesenchymal transition (EMT), while FGF7 knockdown abated the effects. Transcription factor SP1 activated the transcription level of FGF7 and SP1 overexpression aggravated HG-induced SRA01/04 cell injury. SP1 silencing repressed HG-induced SRA01/04 cell viability, migration, invasion and EMT, but these effects were ameliorated by upregulating FGF7. Additionally, SP1 knockdown inhibited the PI3K/AKT pathway by regulating the transcription level of FGF7. CONCLUSION: Transcription factor SP1 activated the transcription level of FGF7 and the PI3K/AKT pathway to regulate HG-induced SRA01/04 cell viability, migration, invasion and EMT.
Subject(s)
Cell Movement , Cell Survival , Epithelial Cells , Epithelial-Mesenchymal Transition , Fibroblast Growth Factor 7 , Glucose , Lens, Crystalline , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Signal Transduction , Sp1 Transcription Factor , Epithelial-Mesenchymal Transition/drug effects , Sp1 Transcription Factor/metabolism , Sp1 Transcription Factor/genetics , Humans , Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Glucose/pharmacology , Epithelial Cells/metabolism , Fibroblast Growth Factor 7/metabolism , Fibroblast Growth Factor 7/genetics , Fibroblast Growth Factor 7/pharmacology , Lens, Crystalline/metabolism , Lens, Crystalline/cytology , Cataract/metabolism , Cells, Cultured , Gene Expression RegulationABSTRACT
Adding 50% vitreous humor to the media surrounding lens explants induces fiber cell differentiation and a significant immune/inflammatory response. While Fgfr loss blocks differentiation in lens epithelial explants, this blockage is partially reversed by deleting Pten. To investigate the functions of the Fgfrs and Pten during lens fiber cell differentiation, we utilized a lens epithelial explant system and conducted RNA sequencing on vitreous humor-exposed explants lacking Fgfrs, or Pten or both Fgfrs and Pten. We found that Fgfr loss impairs both vitreous-induced differentiation and inflammation while the additional loss of Pten restores these responses. Furthermore, transcriptomic analysis suggested that PDGFR-signaling in FGFR-deficient explants is required to mediate the rescue of vitreous-induced fiber differentiation in explants lacking both Fgfrs and Pten. The blockage of ß-crystallin induction in explants lacking both Fgfrs and Pten in the presence of a PDGFR inhibitor supports this hypothesis. Our findings demonstrate that a wide array of genes associated with fiber cell differentiation are downstream of FGFR-signaling and that the vitreous-induced immune responses also depend on FGFR-signaling. Our data also demonstrate that many of the vitreous-induced gene-expression changes in Fgfr-deficient explants are rescued in explants lacking both Fgfrs and Pten.
Subject(s)
Cell Differentiation , Lens, Crystalline , PTEN Phosphohydrolase , PTEN Phosphohydrolase/metabolism , PTEN Phosphohydrolase/genetics , Cell Differentiation/genetics , Animals , Lens, Crystalline/cytology , Lens, Crystalline/metabolism , Mice , Signal Transduction , Gene Expression Profiling , Transcriptome/geneticsABSTRACT
Rationale: Cataract is the leading cause of blindness and low vision worldwide, yet its pathological mechanism is not fully understood. Although macroautophagy/autophagy is recognized as essential for lens homeostasis and has shown potential in alleviating cataracts, its precise mechanism remains unclear. Uncovering the molecular details of autophagy in the lens could provide targeted therapeutic interventions alongside surgery. Methods: We monitored autophagic activities in the lens and identified the key autophagy protein ATG16L1 by immunofluorescence staining, Western blotting, and transmission electron microscopy. The regulatory mechanism of ATG16L1 ubiquitination was analyzed by co-immunoprecipitation and Western blotting. We used the crystal structure of E3 ligase gigaxonin and conducted the docking screening of a chemical library. The effect of the identified compound riboflavin was tested in vitro in cells and in vivo animal models. Results: We used HLE cells and connexin 50 (cx50)-deficient cataract zebrafish model and confirmed that ATG16L1 was crucial for lens autophagy. Stabilizing ATG16L1 by attenuating its ubiquitination-dependent degradation could promote autophagy activity and relieve cataract phenotype in cx50-deficient zebrafish. Mechanistically, the interaction between E3 ligase gigaxonin and ATG16L1 was weakened during this process. Leveraging these mechanisms, we identified riboflavin, an E3 ubiquitin ligase-targeting drug, which suppressed ATG16L1 ubiquitination, promoted autophagy, and ultimately alleviated the cataract phenotype in autophagy-related models. Conclusions: Our study identified an unrecognized mechanism of cataractogenesis involving ATG16L1 ubiquitination in autophagy regulation, offering new insights for treating cataracts.
Subject(s)
Autophagy-Related Proteins , Autophagy , Cataract , Lens, Crystalline , Zebrafish , Animals , Cataract/metabolism , Cataract/drug therapy , Autophagy/drug effects , Autophagy-Related Proteins/metabolism , Lens, Crystalline/metabolism , Lens, Crystalline/drug effects , Humans , Ubiquitination/drug effects , Riboflavin/pharmacology , Disease Models, Animal , Cell LineABSTRACT
A 29-year-old man presented with longstanding, stable, unilateral vision loss in the setting of a remote paintball injury. His examination was notable for a sensory exotropia as well as multiple foci of posterior synechiae, segments of white lenticular material and islands of lenticular cells within a grossly intact capsule, and severe zonular compromise in the affected eye. The majority of the nuclear lens material was absent. The patient was diagnosed with post-traumatic lens absorption and underwent synechialysis, capsulotomy, excision of remnant lenticular material, and placement of a sulcus lens, with significant improvement in visual acuity and ocular alignment following surgery. Our report uniquely highlights identification of a Soemmering's ring in an absorbed lens in the presence of an intact anterior and posterior capsule as well as successful refractive and sensorimotor outcomes following surgical repair despite delay in treatment of many years.
Subject(s)
Eye Injuries , Lens Capsule, Crystalline , Visual Acuity , Humans , Male , Adult , Lens Capsule, Crystalline/surgery , Eye Injuries/diagnosis , Eye Injuries/complications , Eye Injuries/surgery , Eye Injuries/etiology , Lens, Crystalline/injuries , Lens, Crystalline/surgery , Lens Subluxation/etiology , Lens Subluxation/diagnosis , Lens Subluxation/surgeryABSTRACT
PURPOSE: To assess the influence of ocular biometric parameters on intraocular lens (IOL) tilt and decentration after cataract surgery. METHODS: Patients scheduled for cataract surgery were screened for inclusion in this prospective cohort study. Tilt and decentration of the crystalline lens and IOL were measured using the CASIA2 (Tomey). Anterior chamber depth (ACD), lens thickness (LT), and axial length (AL) were preoperatively measured by the IOLMaster 700 (Carl Zeiss Meditec AG). Multivariate regression analysis was performed to assess the influence of ocular biometric parameters on IOL tilt and decentration after cataract surgery. RESULTS: In total, 191 eyes of 120 patients were included. Age was positively correlated with IOL tilt, whereas ACD and AL were negatively correlated with IOL tilt. A strong positive correlation was found between preoperative crystalline lenses and postoperative IOLs in tilt magnitude (r = 0.769, P < .001) and tilt direction (r = 0.688, P < .001). A positive correlation was found between preoperative and postoperative lens decentration magnitude and decentration direction. Greater postoperative IOL tilt and decentration were significantly associated with greater preoperative crystalline lens tilt (P < .001) and decentration (P = .027). CONCLUSIONS: IOL tilt was greater in older patients. Shorter AL and shallower ACD contributed to greater IOL tilt. The tilt and decentration of the IOL will be greater in patients with greater tilt and decentration of the crystalline lens. [J Refract Surg. 2024;40(7):e438-e444.].
Subject(s)
Axial Length, Eye , Biometry , Lens Implantation, Intraocular , Lenses, Intraocular , Phacoemulsification , Humans , Prospective Studies , Male , Female , Aged , Middle Aged , Axial Length, Eye/pathology , Aged, 80 and over , Anterior Chamber/pathology , Artificial Lens Implant Migration/physiopathology , Lens, Crystalline , Visual Acuity/physiology , Adult , Pseudophakia/physiopathologyABSTRACT
Purpose: The purpose of this study was to utilize multi-parametric magnetic resonance imaging (MRI) to investigate in vivo age-related changes in the physiology and optics of mouse lenses where Connexin 50 has been deleted (Cx50KO) or replaced by Connexin 46 (Cx50KI46). Methods: The lenses of transgenic Cx50KO and Cx50KI46 mice were imaged between 3 weeks and 6 months of age using a 7T MRI. Measurements of lens geometry, the T2 (water-bound protein ratios), the refractive index (n), and T1 (free water content) values were calculated by processing the acquired images. The lens power was calculated from an optical model that combined the geometry and the n. All transgenic mice were compared with control mice at the same age. Results: Cx50KO and Cx50KI46 mice developed smaller lenses compared with control mice. The lens thickness, volume, and surface radii of curvatures all increased with age but were limited to the size of the lenses. Cx50KO lenses exhibited higher lens power than Cx50KI46 lenses at all ages, and this was correlated with significantly lower water content in these lenses, which was probably modulated by the gap junction coupling. The refractive power tended to a steady state with age, similar to the control mice. Conclusions: The modification of Cx50 gap junctions significantly impacted lens growth and physiological optics as the mouse aged. The lenses showed delayed development growth, and altered optics governed by different lens physiology. This research provides new insights into how gap junctions regulate the development of the lens's physiological optics.