Experimental eye research最新文献_第7页

Editorial: In memory of Jerry Lutty 社论：纪念杰瑞·卢蒂。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110226

Malia M. Edwards, Robert F. Mullins

引用次数: 0

Estradiol impacts Müller glia and endothelial cell responses in hyperglycemic microenvironments with advanced glycation end products 雌二醇影响mÜller胶质细胞和内皮细胞反应在高血糖微环境与晚期糖基化终产物。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110185

Natalia Castro , Juan S. Peña , Richard Cliver , François Berthiaume , Maribel Vazquez

Diabetic retinopathy is a leading cause of vision loss in working adults, with disproportionate impact on women with lowered estrogen. Sex hormones and their receptors are significant to neuroprotection of the inner blood-retinal barrier (iBRB), a tissue that regulates transport across the neuroretina and vasculature. Moreover, high glucose levels in diabetes lead to the formation of advanced glycation end products (AGEs), which promote inflammation and iBRB breakdown to result in vision loss. This study examined the effects of supplemental estradiol on cell reactivity and cell barrier resistance within an in vitro model of hyperglycemia. Changes in morphology and expression of reactive oxygen species were examined when cells were exposed to a hyperglycemic medium containing AGEs, with and without supplemental estradiol. Cell morphology was assessed via changes in cell area and cell shape index, while intracellular ROS levels were measured using a ROS-sensitive dye. In addition, trans endothelial resistance (TEER) assays were used to measure changes in cell barrier function in response to hyperglycemic conditions, with and without supplemental estradiol. Results show that ROS levels in Müller glia in hyperglycemic conditions significantly decreased in response to supplemental estradiol. The estradiol further increased the resistivity of Müller glia and endothelial cell barriers cultured in high glucose and AGEs. This project illustrates the restorative effects of estradiol in collective responses of cell barriers formed by endothelial cells and Müller glia.

糖尿病性视网膜病变是成年劳动者视力丧失的主要原因，对雌激素水平较低的女性影响更大。性激素及其受体对血液视网膜内屏障（iBRB）的神经保护具有重要意义，iBRB是一种调节神经视网膜和脉管系统运输的组织。此外，糖尿病患者的高葡萄糖水平会导致晚期糖基化终产物（AGEs）的形成，从而促进炎症和iBRB分解，导致视力丧失。本研究在体外高血糖模型中检测了补充雌二醇对细胞反应性和细胞屏障抵抗的影响。当细胞暴露在含有AGEs的高血糖培养基中，添加或不添加雌二醇时，检测了形态和活性氧表达的变化。通过细胞面积和细胞形状指数的变化来评估细胞形态，而使用ROS敏感染料测量细胞内ROS水平。此外，使用反内皮阻力（TEER）试验来测量在有或没有补充雌二醇的情况下，高血糖条件下细胞屏障功能的变化。结果表明，在高血糖状态下，补充雌二醇显著降低了勒神经胶质细胞中的ROS水平。雌二醇进一步增加了高糖和AGEs培养的神经胶质细胞和内皮细胞屏障的电阻率。该项目说明了雌二醇在内皮细胞和勒神经胶质细胞形成的细胞屏障集体反应中的恢复作用。

{"title":"Estradiol impacts Müller glia and endothelial cell responses in hyperglycemic microenvironments with advanced glycation end products","authors":"Natalia Castro , Juan S. Peña , Richard Cliver , François Berthiaume , Maribel Vazquez","doi":"10.1016/j.exer.2024.110185","DOIUrl":"10.1016/j.exer.2024.110185","url":null,"abstract":"<div><div>Diabetic retinopathy is a leading cause of vision loss in working adults, with disproportionate impact on women with lowered estrogen. Sex hormones and their receptors are significant to neuroprotection of the inner blood-retinal barrier (iBRB), a tissue that regulates transport across the neuroretina and vasculature. Moreover, high glucose levels in diabetes lead to the formation of advanced glycation end products (AGEs), which promote inflammation and iBRB breakdown to result in vision loss. This study examined the effects of supplemental estradiol on cell reactivity and cell barrier resistance within an in vitro model of hyperglycemia. Changes in morphology and expression of reactive oxygen species were examined when cells were exposed to a hyperglycemic medium containing AGEs, with and without supplemental estradiol. Cell morphology was assessed via changes in cell area and cell shape index, while intracellular ROS levels were measured using a ROS-sensitive dye. In addition, trans endothelial resistance (TEER) assays were used to measure changes in cell barrier function in response to hyperglycemic conditions, with and without supplemental estradiol. Results show that ROS levels in Müller glia in hyperglycemic conditions significantly decreased in response to supplemental estradiol. The estradiol further increased the resistivity of Müller glia and endothelial cell barriers cultured in high glucose and AGEs. This project illustrates the restorative effects of estradiol in collective responses of cell barriers formed by endothelial cells and Müller glia.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110185"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Increased reactive astrocytes and NLRC4-mediated neuronal pyroptosis in advanced visual structures contralateral to the optic nerve crush eye in mice 小鼠视神经挤压眼对侧高级视觉结构中反应性星形胶质细胞和nlrc4介导的神经元焦亡增加。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2025.110235

Deling Li, Qinyuan Hu, Zongyi Zhan, Xinyi Zhang, Weiting Zeng, Liling Liu, Kaili Wu, Minbin Yu

Currently, research on optic nerve injury predominantly focuses on the retina and optic nerve, but emerging evidence suggests that optic nerve injury also affects advanced visual structures like the superior colliculus (SC) and primary visual cortex (V1 region). However, the exact mechanisms have not been fully explored. This study aims to investigate the characteristics and mechanisms of pathology in the SC and V1 region after optic nerve crush (ONC) to deepen our understanding of the central mechanism of visual injury. After unilateral ONC, visual acuity in the injured eye declined, along with thinning of the retinal nerve fiber layer, and the latency and amplitude of FVEPs decreased. Furthermore, neuronal loss and degeneration were observed in the contralateral SC and V1 region, accompanied by astrocytic activation. Additionally, protein markers C3, and Serping1 for A1 astrocytes, which had neurotoxic effects and S100A10, and PTX3 for A2 astrocytes, which promoted tissue repair, were increased in the two regions. A1 astrocytes were mainly present in the early stages of observation, while A2 astrocytes were mainly increased later. Notably, NLRC4, GSDMD-N, cleaved caspase-1 expression, and IL-1β, IL-18 secretion increased in the contralateral SC and V1 region. Collectively, our findings reveal that A1 (neurotoxic) and A2 astrocytes (neuroprotective), NLRC4-mediated neuronal pyroptosis are enhanced in SC and V1 region contralateral to the ONC eye. The primary visual cortex responds to injury later than the superior colliculus after ONC, with less pronounced damage changes. Reactive astrocytes and NLRC4 inflammasome may act as promising targets for the prevention and treatment of optic nerve injury.

目前，视神经损伤的研究主要集中在视网膜和视神经上，但越来越多的证据表明视神经损伤也影响到高级视觉结构，如上丘（SC）和初级视觉皮层（V1区）。然而，确切的机制尚未得到充分探索。本研究旨在探讨视神经压伤（ONC）后SC和V1区的病理特点和机制，以加深我们对视觉损伤中枢机制的认识。单侧ONC后，损伤眼视力下降，视网膜神经纤维层变薄，fvep潜伏期和振幅降低。此外，在对侧SC和V1区观察到神经元丢失和变性，并伴有星形细胞激活。此外，具有神经毒性作用的A1星形胶质细胞的蛋白标志物C3和Serping1以及促进组织修复的A2星形胶质细胞的蛋白标志物S100A10和PTX3在这两个区域均有所增加。观察早期以A1星形胶质细胞为主，观察后期以A2星形胶质细胞增多为主。NLRC4、GSDMD-N、cleaved caspase-1表达和IL-1β、IL-18分泌在对侧SC和V1区显著升高。总的来说，我们的研究结果表明，A1（神经毒性）和A2星形细胞（神经保护性）、nlrc4介导的神经元焦亡在ONC眼对侧的SC和V1区增强。ONC后初级视觉皮层对损伤的反应晚于上丘，损伤变化不明显。反应性星形胶质细胞和NLRC4炎性体可能是预防和治疗视神经损伤的有希望的靶点。

{"title":"Increased reactive astrocytes and NLRC4-mediated neuronal pyroptosis in advanced visual structures contralateral to the optic nerve crush eye in mice","authors":"Deling Li, Qinyuan Hu, Zongyi Zhan, Xinyi Zhang, Weiting Zeng, Liling Liu, Kaili Wu, Minbin Yu","doi":"10.1016/j.exer.2025.110235","DOIUrl":"10.1016/j.exer.2025.110235","url":null,"abstract":"<div><div>Currently, research on optic nerve injury predominantly focuses on the retina and optic nerve, but emerging evidence suggests that optic nerve injury also affects advanced visual structures like the superior colliculus (SC) and primary visual cortex (V1 region). However, the exact mechanisms have not been fully explored. This study aims to investigate the characteristics and mechanisms of pathology in the SC and V1 region after optic nerve crush (ONC) to deepen our understanding of the central mechanism of visual injury. After unilateral ONC, visual acuity in the injured eye declined, along with thinning of the retinal nerve fiber layer, and the latency and amplitude of FVEPs decreased. Furthermore, neuronal loss and degeneration were observed in the contralateral SC and V1 region, accompanied by astrocytic activation. Additionally, protein markers C3, and Serping1 for A1 astrocytes, which had neurotoxic effects and S100A10, and PTX3 for A2 astrocytes, which promoted tissue repair, were increased in the two regions. A1 astrocytes were mainly present in the early stages of observation, while A2 astrocytes were mainly increased later. Notably, NLRC4, GSDMD-N, cleaved caspase-1 expression, and IL-1β, IL-18 secretion increased in the contralateral SC and V1 region. Collectively, our findings reveal that A1 (neurotoxic) and A2 astrocytes (neuroprotective), NLRC4-mediated neuronal pyroptosis are enhanced in SC and V1 region contralateral to the ONC eye. The primary visual cortex responds to injury later than the superior colliculus after ONC, with less pronounced damage changes. Reactive astrocytes and NLRC4 inflammasome may act as promising targets for the prevention and treatment of optic nerve injury.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110235"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Inhibition of matrix metalloproteases by a chemical cross-linker to halt the corneal degradation in keratoconus 化学交联剂抑制基质金属蛋白酶以阻止圆锥角膜的角膜退化。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110208

Adhithya Subramanian Gopalakrishnan , Sumaiya Sirajudeen , Nasrin Banu , Jessica Nunes , Divya T. Rajendran , Seema Yadav , Namperumalsamy Venkatesh Prajna , Rachel Williams , Dharmalingam Kuppamuthu , Ramprasad Obula Giridhara Gopalan

The need for better and simpler alternative crosslinking strategies to treat keratoconus (KC) is becoming essential as there is only a single approved way to treat it. Recently, conventional UV-A Riboflavin crosslinking is proven to have some disadvantages such as causing damage to the corneal endothelium and inducing keratocyte apoptosis. A chemical cross-linker (CXL) using carbodiimide chemistry and an octanedioic acid spacer is found effective in stiffening the cornea and has the potential to be developed as an alternative therapy to halt KC progression. In order to investigate the molecular changes induced by the cross-linker, we have analyzed the effect of the cross-linker on the activity of matrix metalloproteases (MMPs) in epithelial and stromal layers of KC corneas and in vitro cellular systems to determine its role in stiffening the KC cornea. At well-optimized concentration, KC corneal buttons were treated with the CXL and the stiffening of the cornea was measured. The collagen fibril assembly in the stroma was analyzed using transmission electron microscopy and the activity of MMPs 2 and 9 were visualized using gelatin zymography. KC corneal fibroblasts in culture and tumor necrosis factor-α (TNF-α) induced human corneal epithelial (HCE) cell line were treated with CXL and secretion of MMPs 1, 2, 3 and 9 were analyzed by enzyme-linked immunosorbent assay (ELISA). We found that the CXL stiffened the KC corneas comparable to the normal corneas, with very less cytotoxicity. The collagen fiber assembly was reorganized in an orderly fashion and fibril density and diameter increased after CXL treatment. The activity of MMPs and cathepsin G in the epithelial and stromal layers of KC tissues decreased post-treatment. Secretion and activity of MMPs from the corneal epithelial and stromal cells after CXL treatment were significantly reduced while the epithelial lysyl oxidase activity increased. The CXL, intended to stop the KC progression, modified the extracellular matrix collagen assembly in the stroma and decreased the secretion of a group of metalloproteases and their activity. We have demonstrated a set of molecular changes effected by the CXL, which might aid in the stiffening of the KC cornea.

由于目前只有一种经批准的治疗圆锥角膜的方法，因此需要更好、更简单的交联治疗方法。近年来，传统的UV-A核黄素交联方法被证明存在损伤角膜内皮、诱导角膜细胞凋亡等缺点。一种化学交联剂（CXL）使用碳二亚胺化学和辛烷二酸间隔剂被发现可以有效地硬化角膜，并且有潜力作为一种替代疗法来阻止KC的进展。为了研究交联剂诱导的分子变化，我们分析了交联剂对KC角膜上皮和基质层以及体外细胞系统中基质金属蛋白酶（MMPs）活性的影响，以确定其在KC角膜硬化中的作用。在最佳浓度下，用CXL处理KC角膜按钮，并测量角膜的硬化。用透射电镜分析基质中胶原原纤维的组装，用明胶酶谱法观察MMPs 2和9的活性。用CXL处理培养的KC角膜成纤维细胞和肿瘤坏死因子-α (TNF-α)诱导的人角膜上皮（HCE）细胞株，酶联免疫吸附试验（ELISA）检测MMPs 1、2、3和9的分泌情况。我们发现CXL使KC角膜硬化，与正常角膜相比，细胞毒性非常小。CXL处理后，胶原纤维组装有序重组，纤维密度和直径增加。KC组织上皮和间质层的MMPs和组织蛋白酶G活性降低。CXL处理后，角膜上皮细胞和基质细胞中MMPs的分泌和活性显著降低，而上皮赖氨酸氧化酶活性升高。为了阻止KC的进展，CXL改变了基质中的细胞外基质胶原蛋白组装，降低了一组金属蛋白酶的分泌及其活性。我们已经证明了一组受CXL影响的分子变化，这可能有助于KC角膜的硬化。

{"title":"Inhibition of matrix metalloproteases by a chemical cross-linker to halt the corneal degradation in keratoconus","authors":"Adhithya Subramanian Gopalakrishnan , Sumaiya Sirajudeen , Nasrin Banu , Jessica Nunes , Divya T. Rajendran , Seema Yadav , Namperumalsamy Venkatesh Prajna , Rachel Williams , Dharmalingam Kuppamuthu , Ramprasad Obula Giridhara Gopalan","doi":"10.1016/j.exer.2024.110208","DOIUrl":"10.1016/j.exer.2024.110208","url":null,"abstract":"<div><div>The need for better and simpler alternative crosslinking strategies to treat keratoconus (KC) is becoming essential as there is only a single approved way to treat it. Recently, conventional UV-A Riboflavin crosslinking is proven to have some disadvantages such as causing damage to the corneal endothelium and inducing keratocyte apoptosis. A chemical cross-linker (CXL) using carbodiimide chemistry and an octanedioic acid spacer is found effective in stiffening the cornea and has the potential to be developed as an alternative therapy to halt KC progression. In order to investigate the molecular changes induced by the cross-linker, we have analyzed the effect of the cross-linker on the activity of matrix metalloproteases (MMPs) in epithelial and stromal layers of KC corneas and in vitro cellular systems to determine its role in stiffening the KC cornea. At well-optimized concentration, KC corneal buttons were treated with the CXL and the stiffening of the cornea was measured. The collagen fibril assembly in the stroma was analyzed using transmission electron microscopy and the activity of MMPs 2 and 9 were visualized using gelatin zymography. KC corneal fibroblasts in culture and tumor necrosis factor-α (TNF-α) induced human corneal epithelial (HCE) cell line were treated with CXL and secretion of MMPs 1, 2, 3 and 9 were analyzed by enzyme-linked immunosorbent assay (ELISA). We found that the CXL stiffened the KC corneas comparable to the normal corneas, with very less cytotoxicity. The collagen fiber assembly was reorganized in an orderly fashion and fibril density and diameter increased after CXL treatment. The activity of MMPs and cathepsin G in the epithelial and stromal layers of KC tissues decreased post-treatment. Secretion and activity of MMPs from the corneal epithelial and stromal cells after CXL treatment were significantly reduced while the epithelial lysyl oxidase activity increased. The CXL, intended to stop the KC progression, modified the extracellular matrix collagen assembly in the stroma and decreased the secretion of a group of metalloproteases and their activity. We have demonstrated a set of molecular changes effected by the CXL, which might aid in the stiffening of the KC cornea.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110208"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142834845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mitf over-expression leads to microphthalmia and coloboma in Mitf-cre mice Mitf过表达导致Mitf-cre小鼠小眼症和结肠瘤。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110209

Anne Nathalie Longakit, Hannah Bourget, Catherine D. Van Raamsdonk

The Mitf transcription factor is a critical regulator of the melanocyte lineage and eye development. Mitf activity in different cell types is controlled in part by ten alternative promoters and their resulting isoforms. A useful tool for melanocyte-based research, Mitf-cre was designed to express Cre from the Mitf-M promoter, which is melanocyte specific. However, Mitf-cre mice are also microphthalmic, perhaps because of insertional mutagenesis or disrupted gene expression. Here, we investigated these possibilities and described the eye phenotype. Targeted locus amplification indicated that the transgene integrated on chromosome 2, in between Spred1 and Meis2. The BAC transgene used to make Mitf-cre was larger than expected, carrying three upstream alternative promoters, Mitf-H, Mitf-D, and Mitf-B, which could express their isoforms intact off the transgene. RT-qPCR using eye tissue demonstrated a 5-fold increase in Mitf transcripts containing exon 1B1b, which is shared by Mitf-H, Mitf-D, and Mitf-B, while Spred1 and Meis2 did not differ in their expression. These findings clarify and support the usage of Mitf-cre in conditional mutagenesis in melanocytes. The specific over-expression of these isoforms, which are preferentially expressed in the RPE, presents a unique resource for those interested in eye development and coloboma.

Mitf转录因子是黑素细胞谱系和眼睛发育的关键调节因子。在不同的细胞类型中，Mitf的活性在一定程度上是由十种不同的启动子及其产生的同种异构体控制的。作为一种基于黑素细胞研究的有用工具，Mitf-cre被设计用于表达来自Mitf-M启动子的Cre，这是黑素细胞特异性的。然而，Mitf-cre小鼠也有小眼性，可能是由于插入突变或基因表达中断。在这里，我们研究了这些可能性并描述了眼睛表型。目标基因座扩增表明，该基因整合在2号染色体上，位于Spred1和Meis2之间。用于制造Mitf-cre的BAC转基因比预期的要大，它携带了三个上游替代启动子，Mitf-H、Mitf-D和Mitf-B，它们可以完整地表达它们的同种异构体。使用眼组织的RT-qPCR显示含有外显子1B1b的Mitf转录本增加了5倍，Mitf- h， Mitf- d和Mitf- b共享该转录本，而Spred1和Meis2的表达没有差异。这些发现阐明并支持Mitf-cre在黑素细胞条件诱变中的应用。这些在RPE中优先表达的同种异构体的特异性过表达，为那些对眼睛发育和结肠瘤感兴趣的人提供了独特的资源。

{"title":"Mitf over-expression leads to microphthalmia and coloboma in Mitf-cre mice","authors":"Anne Nathalie Longakit, Hannah Bourget, Catherine D. Van Raamsdonk","doi":"10.1016/j.exer.2024.110209","DOIUrl":"10.1016/j.exer.2024.110209","url":null,"abstract":"<div><div>The Mitf transcription factor is a critical regulator of the melanocyte lineage and eye development. Mitf activity in different cell types is controlled in part by ten alternative promoters and their resulting isoforms. A useful tool for melanocyte-based research, <em>Mitf-cre</em> was designed to express Cre from the Mitf-M promoter, which is melanocyte specific. However, <em>Mitf-cre</em> mice are also microphthalmic, perhaps because of insertional mutagenesis or disrupted gene expression. Here, we investigated these possibilities and described the eye phenotype. Targeted locus amplification indicated that the transgene integrated on chromosome 2, in between <em>Spred1</em> and <em>Meis2</em>. The BAC transgene used to make <em>Mitf-cre</em> was larger than expected, carrying three upstream alternative promoters, Mitf-H, Mitf-D, and Mitf-B, which could express their isoforms intact off the transgene. RT-qPCR using eye tissue demonstrated a 5-fold increase in <em>Mitf</em> transcripts containing exon 1B1b, which is shared by Mitf-H, Mitf-D, and Mitf-B, while <em>Spred1</em> and <em>Meis2</em> did not differ in their expression. These findings clarify and support the usage of <em>Mitf-cre</em> in conditional mutagenesis in melanocytes. The specific over-expression of these isoforms, which are preferentially expressed in the RPE, presents a unique resource for those interested in eye development and coloboma.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110209"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effects of atropine on choroidal hemodynamics and VEGFA and HIF-1α expression in form-deprivation myopia guinea pigs 阿托品对形态剥夺性近视豚鼠脉络膜血流动力学及VEGFA和HIF-1α表达的影响

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110214

Danyang Che , Danlei Qiao , Lingfeng Lv , Yingjie Zhang , Yiting Cao , Fang Li , Shanbao Tong , Peng Miao , Jibo Zhou

We investigated the mechanism of action of atropine in myopia control by examining its effect on choroidal hemodynamics. Blood flow was evaluated using indocyanine green (ICG) fluorescence and molecular variation during the development of form-deprivation myopia (FDM) and atropine treatment in guinea pigs. Guinea pigs were divided randomly into the normal control (NC), FDM, and FDM + 1% atropine (ATR) groups, and evaluated by spherical equivalent refractive error (SE) and axial length (AL). Choroidal hemodynamic parameters were measured via ICG fluorescence imaging including the maximal ICG fluorescence intensity (I_max), rising time (T_rising), blood flow index (BFI), and mean transit time (MTT). Additionally, the expression in the choroid-RPE complex of choroidal vascular endothelial growth factor A (VEGFA) and HIF-1 α were assessed via Western blotting. Atropine inhibited the development of FDM, with effects of FD on both SE and AL being reduced. ICG fluorescence hemodynamic wide-field maps and time-series curves revealed that the atropine significantly accelerated choroidal blood flow, with reduced T_rising and MTT, while increasing I_max, BFI and the number of lobulated choriocapillaris structures compared with the FDM group. In terms of molecular markers, atropine inhibited the effect of FDM, increasing VEGFA levels and reducing HIF-1α expression. These findings suggest that atropine improved choroidal hemodynamics and changed vascular markers, potentially contributing to its role in inhibiting the progression of myopia in the FDM model.

我们通过观察阿托品对脉络膜血流动力学的影响来探讨其控制近视的作用机制。采用吲哚菁绿（ICG）荧光和阿托品治疗评估豚鼠形态剥夺性近视（FDM）发展过程中的血流变化。将豚鼠随机分为正常对照组（NC）、FDM组和FDM + 1%阿托品组（ATR），采用球等效屈光差（SE）和眼轴长度（AL）进行评价。通过ICG荧光成像测量脉络膜血流动力学参数，包括最大ICG荧光强度（Imax）、上升时间（Trising）、血流指数（BFI）和平均传递时间（MTT）。Western blotting检测脉络膜血管内皮生长因子A （VEGFA）和HIF-1 α在脉络膜- rpe复合体中的表达。阿托品抑制FDM的发展，FD对SE和AL的影响减弱。ICG荧光血流动力学宽场图和时间序列曲线显示，与FDM组相比，阿托品显著加速脉络膜血流，降低Trising和MTT，同时增加Imax、BFI和分叶绒毛膜毛细血管结构的数量。在分子标记方面，阿托品抑制FDM的作用，增加VEGFA水平，降低HIF-1α表达。这些发现表明，阿托品改善脉络膜血流动力学和改变血管标志物，可能有助于其在抑制FDM模型近视进展中的作用。

{"title":"Effects of atropine on choroidal hemodynamics and VEGFA and HIF-1α expression in form-deprivation myopia guinea pigs","authors":"Danyang Che , Danlei Qiao , Lingfeng Lv , Yingjie Zhang , Yiting Cao , Fang Li , Shanbao Tong , Peng Miao , Jibo Zhou","doi":"10.1016/j.exer.2024.110214","DOIUrl":"10.1016/j.exer.2024.110214","url":null,"abstract":"<div><div>We investigated the mechanism of action of atropine in myopia control by examining its effect on choroidal hemodynamics. Blood flow was evaluated using indocyanine green (ICG) fluorescence and molecular variation during the development of form-deprivation myopia (FDM) and atropine treatment in guinea pigs. Guinea pigs were divided randomly into the normal control (NC), FDM, and FDM + 1% atropine (ATR) groups, and evaluated by spherical equivalent refractive error (SE) and axial length (AL). Choroidal hemodynamic parameters were measured via ICG fluorescence imaging including the maximal ICG fluorescence intensity (<em>I</em><sub>max</sub>), rising time (<em>T</em><sub>rising</sub>), blood flow index (BFI), and mean transit time (MTT). Additionally, the expression in the choroid-RPE complex of choroidal vascular endothelial growth factor A (VEGFA) and HIF-1 α were assessed via Western blotting. Atropine inhibited the development of FDM, with effects of FD on both SE and AL being reduced. ICG fluorescence hemodynamic wide-field maps and time-series curves revealed that the atropine significantly accelerated choroidal blood flow, with reduced <em>T</em><sub>rising</sub> and MTT, while increasing <em>I</em><sub>max</sub>, BFI and the number of lobulated choriocapillaris structures compared with the FDM group. In terms of molecular markers, atropine inhibited the effect of FDM, increasing VEGFA levels and reducing HIF-1α expression. These findings suggest that atropine improved choroidal hemodynamics and changed vascular markers, potentially contributing to its role in inhibiting the progression of myopia in the FDM model.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110214"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142871860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of the IL-6 trans-signaling pathway in the absence or presence of TGF-β2 on Schlemm's canal endothelial cells TGF-β2缺失或存在时IL-6反式信号通路对施莱姆管内皮细胞的影响

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110215

Mai Urahashi , Tomokazu Fujimoto , Miyuki Inoue-Mochita , Toshihiro Inoue

Intraocular pressure (IOP) is regulated through the balance of production and drainage of aqueous humor. The main route of aqueous-humor outflow comprises the trabecular meshwork (TM) and Schlemm's canal (SC). We reported that IL-6 trans-signaling can inhibit TGF-β signaling in TM cells and may affect regulation of IOP. However, the function of IL-6 trans-signaling in SC cells remains unclear. Therefore, we investigated the role of IL-6 trans-signaling in monkey SC cells. Simultaneous treatment with IL-6 and soluble IL-6 receptor (sIL-6R) significantly decreased the trans-endothelial electrical resistance (TER) of SC cells and reduced aqueous-humor outflow resistance. Moreover, activation of IL-6 trans-signaling significantly reduced expression of fibronectin, ZO-1 and claudin-5, and increased that of several matrix metalloproteinases. We also investigated the effect of IL-6 trans-signaling on TGF-β2-induced changes in SC cells. Simultaneous treatment with IL-6 and sIL-6R significantly suppressed the TGF-β2-induced increase in the TER of SC cells but did not affect the activity of the TGF-β2 signaling pathway. By contrast, the TGF-β2-induced increases in the expression of fibronectin and collagen type I were significantly decreased upon simultaneous treatment with IL-6 and sIL-6R. The results show that IL-6 trans-signaling suppressed TGF-β2-induced increase in outflow resistance.

眼内压（IOP）是通过房水生成和排出的平衡来调节的。房水的主要流出途径包括小梁网（TM）和施莱姆管（SC）。我们报道了IL-6反式信号传导可以抑制TM细胞中TGF-β信号传导，并可能影响IOP的调节。然而，IL-6反式信号在SC细胞中的功能尚不清楚。因此，我们研究了IL-6反式信号在猴SC细胞中的作用。同时使用IL-6和可溶性IL-6受体（sIL-6R）可显著降低SC细胞的跨内皮电阻（TER）和房水流出阻力。此外，IL-6反式信号的激活显著降低了纤维连接蛋白、ZO-1和cladin -5的表达，并增加了几种基质金属蛋白酶的表达。我们还研究了IL-6反式信号通路对TGF-β2诱导的SC细胞变化的影响。IL-6和sIL-6R同时处理可显著抑制TGF-β2诱导的SC细胞TER升高，但不影响TGF-β2信号通路的活性。而IL-6和sIL-6R同时治疗时，TGF-β2诱导的纤维连接蛋白和I型胶原表达的升高明显降低。结果表明，IL-6反式信号通路抑制TGF-β2诱导的流出阻力增加。

{"title":"Effect of the IL-6 trans-signaling pathway in the absence or presence of TGF-β2 on Schlemm's canal endothelial cells","authors":"Mai Urahashi , Tomokazu Fujimoto , Miyuki Inoue-Mochita , Toshihiro Inoue","doi":"10.1016/j.exer.2024.110215","DOIUrl":"10.1016/j.exer.2024.110215","url":null,"abstract":"<div><div>Intraocular pressure (IOP) is regulated through the balance of production and drainage of aqueous humor. The main route of aqueous-humor outflow comprises the trabecular meshwork (TM) and Schlemm's canal (SC). We reported that IL-6 <em>trans</em>-signaling can inhibit TGF-β signaling in TM cells and may affect regulation of IOP. However, the function of IL-6 <em>trans</em>-signaling in SC cells remains unclear. Therefore, we investigated the role of IL-6 <em>trans</em>-signaling in monkey SC cells. Simultaneous treatment with IL-6 and soluble IL-6 receptor (sIL-6R) significantly decreased the <em>trans</em>-endothelial electrical resistance (TER) of SC cells and reduced aqueous-humor outflow resistance. Moreover, activation of IL-6 <em>trans</em>-signaling significantly reduced expression of fibronectin, ZO-1 and claudin-5, and increased that of several matrix metalloproteinases. We also investigated the effect of IL-6 <em>trans</em>-signaling on TGF-β2-induced changes in SC cells. Simultaneous treatment with IL-6 and sIL-6R significantly suppressed the TGF-β2-induced increase in the TER of SC cells but did not affect the activity of the TGF-β2 signaling pathway. By contrast, the TGF-β2-induced increases in the expression of fibronectin and collagen type I were significantly decreased upon simultaneous treatment with IL-6 and sIL-6R. The results show that IL-6 <em>trans</em>-signaling suppressed TGF-β2-induced increase in outflow resistance.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110215"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genetic variants in PIKFYVE: A review of ocular phenotypes PIKFYVE的遗传变异：眼部表型研究综述。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110211

Ehsan Misaghi , Peter Kannu , Ian M. MacDonald , Matthew D. Benson

Many studies have identified disease-causing variants of PIKFYVE in ocular tissues; however, a comprehensive review of these variants and their ocular phenotypes is lacking. The phosphoinositide kinase PIKFYVE plays crucial roles in the endolysosomal pathway in autophagy and phagocytosis, both essential for cellular homeostasis. In this review, we evaluate the reported disease-causing PIKFYVE variants and their associated phenotypes in humans to identify potential genotype-phenotype correlations. Variants in PIKFYVE have been associated with corneal fleck dystrophy, congenital cataracts and possibly keratoconus. There are unvalidated associations of variants in PIKFYVE with autism spectrum disorder and congenital heart disease. We show that variants causing corneal fleck dystrophy exist in the chaperonin-like domain of PIKFYVE as well as the region between the chaperonin-like and the kinase domains. Similarly, congenital cataract variants appear to be specific to the kinase domain of the protein. This review consolidates existing knowledge on PIKFYVE variants in ocular disease and bridges fundamental science and clinical manifestations, potentially informing future diagnostic and treatment strategies for PIKFYVE-associated ocular disorders.

许多研究已经确定了眼部组织中PIKFYVE的致病变异；然而，缺乏对这些变异及其眼部表型的全面回顾。磷酸肌肽激酶PIKFYVE在自噬和吞噬的内溶酶体途径中起着至关重要的作用，这两个途径都是细胞稳态所必需的。在这篇综述中，我们评估了已报道的致病PIKFYVE变异及其在人类中的相关表型，以确定潜在的基因型-表型相关性。PIKFYVE的变异与角膜斑点营养不良、先天性白内障和可能的圆锥角膜有关。PIKFYVE变异与自闭症谱系障碍和先天性心脏病之间存在未经证实的关联。我们发现，导致角膜斑点营养不良的变异存在于PIKFYVE的伴侣蛋白样结构域以及伴侣蛋白样结构域和激酶结构域之间的区域。同样，先天性白内障变异似乎是特异性的激酶结构域的蛋白质。本综述巩固了关于PIKFYVE变异在眼部疾病中的现有知识，并将基础科学与临床表现联系起来，可能为未来PIKFYVE相关眼部疾病的诊断和治疗策略提供信息。

{"title":"Genetic variants in PIKFYVE: A review of ocular phenotypes","authors":"Ehsan Misaghi , Peter Kannu , Ian M. MacDonald , Matthew D. Benson","doi":"10.1016/j.exer.2024.110211","DOIUrl":"10.1016/j.exer.2024.110211","url":null,"abstract":"<div><div>Many studies have identified disease-causing variants of <em>PIKFYVE</em> in ocular tissues; however, a comprehensive review of these variants and their ocular phenotypes is lacking. The phosphoinositide kinase PIKFYVE plays crucial roles in the endolysosomal pathway in autophagy and phagocytosis, both essential for cellular homeostasis. In this review, we evaluate the reported disease-causing <em>PIKFYVE</em> variants and their associated phenotypes in humans to identify potential genotype-phenotype correlations. Variants in <em>PIKFYVE</em> have been associated with corneal fleck dystrophy, congenital cataracts and possibly keratoconus. There are unvalidated associations of variants in <em>PIKFYVE</em> with autism spectrum disorder and congenital heart disease. We show that variants causing corneal fleck dystrophy exist in the chaperonin-like domain of PIKFYVE as well as the region between the chaperonin-like and the kinase domains. Similarly, congenital cataract variants appear to be specific to the kinase domain of the protein. This review consolidates existing knowledge on <em>PIKFYVE</em> variants in ocular disease and bridges fundamental science and clinical manifestations, potentially informing future diagnostic and treatment strategies for PIKFYVE-associated ocular disorders.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110211"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853408","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Exploring ocular disorders in Parkinson's disease: A comprehensive review and future perspectives 帕金森氏病眼部疾病的研究：综述与展望

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110225

Minal Thacker , Ka Ying Wong , Liping Zhou , Juewen Liu , Man-Sau Wong

Parkinson's disease (PD) is a multifaceted neurodegenerative disorder characterized by predominantly motor symptoms. However, recent research has broadened our understanding of PD by revealing its impact on non-motor functions, including ocular manifestations. This review explored the intricate relationship between PD and ocular health, shedding light on the mechanisms underlying common ocular diseases such as dry eye disease, cataract, glaucoma, age-related macular degeneration, and diabetic retinopathy. It also underscores the importance of recognizing ocular manifestations as potential early markers of PD, as well as their impact on patients' daily activities, necessitating prompt identification and intervention to prevent complications and enhance the overall quality of life. Furthermore, future research should prioritize unraveling the potential association between PD and other prevalent ocular diseases, such as myopia, to formulate effective treatment strategies.

帕金森病（PD）是一种以运动症状为主的多方面神经退行性疾病。然而，最近的研究通过揭示PD对包括眼部表现在内的非运动功能的影响，拓宽了我们对PD的理解。本文探讨了PD与眼部健康之间的复杂关系，揭示了干眼病、白内障、青光眼、年龄相关性黄斑变性和糖尿病视网膜病变等常见眼部疾病的发病机制。它还强调了将眼部表现视为PD的潜在早期标志的重要性，以及它们对患者日常活动的影响，需要及时识别和干预，以预防并发症并提高整体生活质量。此外，未来的研究应优先揭示PD与其他常见眼部疾病（如近视）之间的潜在关联，以制定有效的治疗策略。

引用次数: 0

SP1/COL1A2/ZEB1 axis promotes TGF-β2-induced lens epithelial cell proliferation, migration, invasion and EMT process SP1/COL1A2/ZEB1轴促进TGF-β2诱导的晶状体上皮细胞增殖、迁移、侵袭和EMT过程。

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-02-01 DOI: 10.1016/j.exer.2024.110220

Lili Zhao , Ping Wang , Lianyi Sun, Weimei Ma, Lei Yu

Posterior capsule opacification (PCO) is the most common complication after cataract surgery. In this study, we used transforming growth factor beta-2 (TGF-β2)-induced SRA01/04 cells to mimic PCO cell model and explored the functions and underlying mechanisms of specific protein 1 (SP1) in TGF-β2-induced SRA01/04 cell development. MTT assay and EdU assay were carried out to explore the proliferation of SRA01/04 cells. Transwell assay and wound-healing assay were performed to investigate SRA01/04 cell migration and invasion. Chromatin Immunoprecipitation (ChIP) assay, dual-luciferase reporter assay and Co-immunoprecipitation (Co-IP) assay were used to analyze the relations of SP1, COL1A2 and ZEB1. TGF-β2 treatment led to the promotion of SRA01/04 cell proliferation, migration, invasion and EMT process. COL1A2 level was induced by TGF-β2 treatment and COL1A2 knockdown inhibited TGF-β2-induced SRA01/04 cell proliferation, migration, invasion and EMT. SP1 could activate the transcription of COL1A2. SP1 overexpression promoted TGF-β2-induced SRA01/04 cell injury by regulating COL1A2 expression. Moreover, COL1A2 interacted with ZEB1 and COL1A2 knockdown-mediated effects on the proliferation, migration, invasion and EMT of TGF-β2-induced SRA01/04 cells were abrogated by elevating ZEB1. SP1 regulated COL1A2 and then mediated ZEB1 to affect the proliferation, migration, invasion and EMT of TGF-β2-induced SRA01/04 cells.

后囊膜混浊是白内障术后最常见的并发症。本研究采用转化生长因子β -2 （TGF-β2）诱导的SRA01/04细胞模拟PCO细胞模型，探讨特异性蛋白1 （SP1）在TGF-β2诱导的SRA01/04细胞发育中的作用及其机制。采用MTT法和EdU法观察SRA01/04细胞的增殖情况。Transwell法和创面愈合法观察SRA01/04细胞的迁移和侵袭。采用染色质免疫沉淀（ChIP）法、双荧光素酶报告基因法和共免疫沉淀（Co-IP）法分析SP1、COL1A2与ZEB1的关系。TGF-β2处理可促进SRA01/04细胞增殖、迁移、侵袭和EMT过程。TGF-β2处理可诱导COL1A2水平，COL1A2敲低可抑制TGF-β2诱导的SRA01/04细胞增殖、迁移、侵袭和EMT。SP1可以激活COL1A2的转录水平。SP1过表达通过调节COL1A2表达促进TGF-β2诱导的SRA01/04细胞损伤。此外，COL1A2与ZEB1相互作用，COL1A2敲低介导的对TGF-β2诱导的SRA01/04细胞增殖、迁移、侵袭和EMT的影响通过上调ZEB1而被消除。SP1通过调控COL1A2，进而介导ZEB1影响TGF-β2诱导的SRA01/04细胞的增殖、迁移、侵袭和EMT。

{"title":"SP1/COL1A2/ZEB1 axis promotes TGF-β2-induced lens epithelial cell proliferation, migration, invasion and EMT process","authors":"Lili Zhao , Ping Wang , Lianyi Sun, Weimei Ma, Lei Yu","doi":"10.1016/j.exer.2024.110220","DOIUrl":"10.1016/j.exer.2024.110220","url":null,"abstract":"<div><div>Posterior capsule opacification (PCO) is the most common complication after cataract surgery. In this study, we used transforming growth factor beta-2 (TGF-β2)-induced SRA01/04 cells to mimic PCO cell model and explored the functions and underlying mechanisms of specific protein 1 (SP1) in TGF-β2-induced SRA01/04 cell development. MTT assay and EdU assay were carried out to explore the proliferation of SRA01/04 cells. Transwell assay and wound-healing assay were performed to investigate SRA01/04 cell migration and invasion. Chromatin Immunoprecipitation (ChIP) assay, dual-luciferase reporter assay and Co-immunoprecipitation (Co-IP) assay were used to analyze the relations of SP1, COL1A2 and ZEB1. TGF-β2 treatment led to the promotion of SRA01/04 cell proliferation, migration, invasion and EMT process. COL1A2 level was induced by TGF-β2 treatment and COL1A2 knockdown inhibited TGF-β2-induced SRA01/04 cell proliferation, migration, invasion and EMT. SP1 could activate the transcription of COL1A2. SP1 overexpression promoted TGF-β2-induced SRA01/04 cell injury by regulating COL1A2 expression. Moreover, COL1A2 interacted with ZEB1 and COL1A2 knockdown-mediated effects on the proliferation, migration, invasion and EMT of TGF-β2-induced SRA01/04 cells were abrogated by elevating ZEB1. SP1 regulated COL1A2 and then mediated ZEB1 to affect the proliferation, migration, invasion and EMT of TGF-β2-induced SRA01/04 cells.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"251 ","pages":"Article 110220"},"PeriodicalIF":3.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0