Investigative ophthalmology & visual science最新文献

{"title":"Nicotinamide N-Methyltransferase in the Inflammatory Pathogenesis of Graves' Orbitopathy.","authors":"Dayoon Cho, Soo Hyun Choi, Jin Sook Yoon, JaeSang Ko","doi":"10.1167/iovs.66.5.3","DOIUrl":"https://doi.org/10.1167/iovs.66.5.3","url":null,"abstract":"<p><strong>Purpose: </strong>Nicotinamide N-methyltransferase (NNMT) has been implicated in inflammatory autoimmune disease pathogenesis, although its pro-inflammatory role in Graves' orbitopathy (GO) is unclear. Therefore, we investigated the influence and mechanisms of NNMT in GO inflammation.</p><p><strong>Methods: </strong>We evaluated NNMT mRNA expression in GO and non-GO orbital tissues via reverse transcription-quantitative PCR analysis. A pro-inflammatory process was induced in primary cultured orbital fibroblasts via interleukin (IL)-1β treatment, and NNMT expression was assessed by Western blotting. To further investigate the role of NNMT in GO inflammation, we inhibited NNMT expression and activity using small interfering RNA (siRNA) and pharmacologic antagonists, respectively. The production of inflammatory cytokines and intracellular signaling molecules were analyzed via Western blotting and enzyme-linked immunosorbent assay analysis.</p><p><strong>Results: </strong>NNMT mRNA expression levels were higher in GO orbital tissues than in healthy orbital tissues. Tissues from patients with type Ⅱ GO showed higher NNMT expression than those with type Ⅰ GO. Pro-inflammatory stimulation induced NNMT expression in dose- and time-dependent manners. NNMT siRNA and antagonists attenuated the expression of pro-inflammatory cytokines (IL-6, IL-8, and monocyte chemotactic protein-1), cyclooxygenase-2, and prostaglandin E2 in orbital fibroblasts. NNMT silencing downregulated the active forms of intracellular signaling molecules (extracellular signal-regulated kinase, c-Jun-terminal kinase, and p38).</p><p><strong>Conclusions: </strong>Our results demonstrate that NNMT was associated with the inflammatory mechanisms of GO. Inhibiting NNMT, either through mRNA silencing or pharmacologic antagonism, markedly reduced pro-inflammatory reactions. These findings suggest that targeting NNMT is a promising therapeutic strategy for managing inflammation in GO.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"3"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12054684/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144005533","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}

{"title":"Cholesterol and Q147E Deamidation Modulates αA-Crystallin Membrane Binding Elucidating Protective Role of Lens Membrane Composition Changes With Aging.","authors":"Preston Hazen, Nawal K Khadka, Laxman Mainali","doi":"10.1167/iovs.66.5.8","DOIUrl":"https://doi.org/10.1167/iovs.66.5.8","url":null,"abstract":"<p><strong>Purpose: </strong>The αA-Crystallin (αAc) binding with lens membranes increases with age and cataract formation. However, the role of lipids and cholesterol (Chol) in Q147E-αAc membrane binding remains unclear, which we aim to elucidate in this study.</p><p><strong>Methods: </strong>We have used the electron paramagnetic resonance spin-labeling method to probe the Chol/ 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) and Chol/ sphingomyelin (SM) membranes binding with wild-type (WT) and Q147E-αAc.</p><p><strong>Results: </strong>Compared to WT-αAc, the Q147E mutant had increased binding to POPC and decreased binding to SM membranes without Chol. Adding 33 mol% Chol to the POPC and SM membranes decreased the binding of WT and, to a lesser degree, decreased the binding of Q147E-αAc to the membranes. Adding 60 mol% Chol completely inhibited Q147E mutant and WT binding to POPC membranes. However, 33 and 60 mol% Chol completely inhibited WT and Q147E mutant binding to SM membranes, respectively. WT and Q147E-αAc membrane binding decreased membrane mobility while increasing order and hydrophobicity near the headgroup.</p><p><strong>Conclusions: </strong>In Chol-free membranes, the deamidated Q147E-αAc binds significantly more to the POPC membranes compared to WT, whereas WT binds significantly more to the SM membranes compared to Q147E-αAc. In contrast, for 33 mol% Chol-containing membranes, the deamidated Q147E-αAc binds significantly more to POPC and SM membranes than WT. Conversely, 60 mol% Chol-containing membranes completely inhibit WT and deamidated Q147E-αAc binding to POPC and SM membranes. These results suggest that increased Chol content of the lens membranes during aging protects against accumulation of modified proteins on the membrane associated with cataracts.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"8"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12060060/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008985","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}

{"title":"Aqueous Humor Levels of Vascular Endothelial Growth Factor in Patients With Dry Age-Related Macular Degeneration and Subretinal Drusenoid Deposits.","authors":"Eun Gyu Yoon, Ki Tae Nam, Mihyun Choi, Kwang-Eon Choi, Cheolmin Yun","doi":"10.1167/iovs.66.5.10","DOIUrl":"https://doi.org/10.1167/iovs.66.5.10","url":null,"abstract":"<p><strong>Purpose: </strong>We sought to investigate aqueous humor levels of growth factors and cytokines related to human angiogenesis in patients with dry age-related macular degeneration (AMD).</p><p><strong>Methods: </strong>This prospective study classified patients with dry AMD into two groups of patients-those with soft drusen and those with both soft drusen and subretinal drusenoid deposits (SDDs). Aqueous humor samples were collected from each group and from a control group to analyze intraocular cytokine concentrations and examine their associations with AMD characteristics.</p><p><strong>Results: </strong>A total of 48 participants, 16 per group, were enrolled in the study. The vascular endothelial growth factor (VEGF)-A level was highest in the soft drusen with SDD group (229.21 ± 88.26 pg/mL) compared to the soft drusen group (167.54 ± 92.71 pg/mL) and the control group (140.73 ± 84.91 pg/mL) (P = 0.021). There were no significant differences in the concentrations of angiopoietin-2, placental growth factor, interleukin-1α, interleukin-1β, interleukin-6, interleukin-10, or tumor necrosis factor-α among the groups (all P > 0.05). In the soft drusen with SDD group, a higher cube root of drusen volume (β = 0.533, P = 0.033) was significantly associated with an elevated VEGF-A level.</p><p><strong>Conclusions: </strong>In eyes with dry AMD, those with both soft drusen and SDDs exhibited higher intraocular VEGF-A levels than those with only soft drusen, and these levels correlated with the cube root of drusen volume.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"10"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12060072/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002392","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}

{"title":"Complement C3/C3aR Signaling Pathway Inhibition Ameliorates Retinal Damage in Experimental Retinal Vein Occlusion.","authors":"Yanying Zhao, Zhengwei Ge, Tingting Guo, Hengwei Liu, Yufan Zhou, Juan Chen, Heping Xu, Zhongping Chen","doi":"10.1167/iovs.66.5.2","DOIUrl":"https://doi.org/10.1167/iovs.66.5.2","url":null,"abstract":"<p><strong>Purpose: </strong>Retinal vein occlusion (RVO) is a common retinal vascular disease that severely threatens visual function. This study aims to elucidate the role of the complement C3/C3aR signaling pathway in a laser-induced RVO mouse model and to explore its potential as a therapeutic target.</p><p><strong>Methods: </strong>RVO was induced in C57BL/6J mice using laser photocoagulation combined with photosensitizer dye administration. Two days later, retinal tissues were collected for bulk RNA sequencing. The activation of the C3/C3aR signaling pathway was validated through RT-qPCR and Western blot. The C3aR antagonist SB290157 (C3aRA) was administered intravitreally and retinal morphological and functional changes were examined 1, 2, and 8 days later by optical coherence tomography (OCT), fundus photography (FP), and fluorescein angiography (FA), optomotor response (OKR) test, and electroretinogram (ERG).</p><p><strong>Results: </strong>RVO mice exhibited marked increases in retinal thickness (P < 0.001) and fluorescence leakage (P < 0.01) compared to the sham-laser group. Bulk RNA-seq revealed significant upregulation of the complement pathway. Elevated expression of C3 and C3aR (P < 0.05) was confirmed by RT-qPCR and Western blot. Blocking C3aR with SB290157 significantly alleviated RVO-induced retinal edema, vascular leakage, and structural damage. Functional assessment showed that SB290157 treatment significantly improved contrast sensitivity (P < 0.05), increased b-wave (P < 0.001), and oscillatory potentials (Ops) amplitudes (P < 0.05) in RVO mice. RNA-seq analysis demonstrated that SB290157 significantly reduced the inflammatory mediator-related pathways and upregulated visual perception pathways (P < 0.05).</p><p><strong>Conclusions: </strong>The complement C3/C3aR signaling pathway is critically involved in RVO-induced retinal damage and targeting this pathway may be a promising approach for RVO treatment.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"2"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12054687/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144024249","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}

{"title":"Regulatory Effects of the Wnt7b/β-Catenin/MMP-2 Signaling Pathway on Scleral Stiffness in Guinea Pigs With Form-Deprivation Myopia.","authors":"Binyu Wen, Hangyu Li, Hui Tao, Hong Ren, Bosheng Ma, Mengdi Shi, Shen Chen, Jiaqi Du, Ziyi Cai, Jing Zhang, Dongshi Guan, Zhihong Deng","doi":"10.1167/iovs.66.5.19","DOIUrl":"https://doi.org/10.1167/iovs.66.5.19","url":null,"abstract":"<p><strong>Purpose: </strong>The development and progression of myopia are influenced by the Wnt7b/β-catenin signaling pathway. This study investigated the specific impacts of this pathway on the biomechanical properties of the sclera by altering the expression of matrix metalloproteinase-2 (MMP-2) to regulate type I collagen (collagen I) levels.</p><p><strong>Methods: </strong>We examined the effects of the Wnt7b/β-catenin signaling pathway and MMP-2 on human fetal scleral fibroblasts (HFSFs) and the sclera of guinea pigs with form-deprivation myopia (FDM). To explore the effects of the Wnt7b/β-catenin pathway and the role of MMP-2 in this context, we treated HFSFs and guinea pig sclera with specific agonists and inhibitors targeting Wnt7b/β-catenin and MMP-2. The expression levels of Wnt7b, MMP-2, and collagen I were subsequently analyzed quantitatively via western blot (WB) analysis, immunofluorescence, and quantitative real-time PCR (qRT-PCR) to assess protein and mRNA changes in response to pathway manipulation. Atomic force microscopy (AFM) was used to measure the elastic modulus of the treated HFSFs and guinea pig sclera to directly evaluate changes in cell and tissue stiffness. In the FDM model, essential ocular parameters such as refractive error and axial length (AL) were also assessed.</p><p><strong>Results: </strong>In vivo and in vitro activation of the Wnt7b/β-catenin signaling pathway significantly upregulated MMP-2 expression, which was accompanied by a notable decrease in collagen I levels. This change led to a reduction in the elastic modulus of both HFSFs and the sclera of guinea pigs with FDM. These significant biomechanical changes in the scleral tissue were indicated by a reduction in stiffness. Alterations in scleral biomechanics were associated with changes in ocular parameters, including an increase in AL and a myopic shift in refraction.</p><p><strong>Conclusions: </strong>The Wnt7b/β-catenin pathway regulates scleral biomechanics by upregulating MMP-2 expression, which leads to increased collagen I degradation and, consequently, an increase in axial elongation and a myopic shift in refractive error.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"19"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12068525/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143990081","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}

{"title":"LncRNA HOTAIR Interaction With WTAP Promotes m6A Methyltransferase Complex Assembly and Posterior Capsule Opacification Formation by Increasing THBS1.","authors":"Xi Chen, Chenshuang Li, Jiankui Li, Zaoxia Guo, Siqi Zhang, Chenjun Guo, Hong Yan","doi":"10.1167/iovs.66.5.20","DOIUrl":"https://doi.org/10.1167/iovs.66.5.20","url":null,"abstract":"<p><strong>Purpose: </strong>To explore the role of long non-coding RNAs (lncRNAs) and N6-methyladenosine (m6A) in posterior capsule opacification (PCO) and their underlying mechanisms.</p><p><strong>Methods: </strong>The localization of lncRNAs and proteins was analyzed using fluorescence in situ hybridization and immunofluorescence staining. RNA m6A quantification, RNA immunoprecipitation, co-immunoprecipitation, MeRIP-seq, MeRIP-qPCR, western blotting, wound healing, and Transwell assays were applied to elucidate the underlying mechanisms.</p><p><strong>Results: </strong>The levels of lncRNA HOX transcript antisense intergenic RNA (HOTAIR) and m6A methylation increased significantly during epithelial-mesenchymal transition (EMT) in lens epithelial cells (LECs). HOTAIR promoted EMT and m6A methyltransferase activity but had no effect on methyltransferase activity and was not modified by m6A. Nevertheless, HOTAIR interacted with WT1-associated protein (WTAP), a key m6A writer protein, facilitating WTAP-mediated recruitment of METTL3-METTL14 heterodimers and enhancing m6A modification. The HOTAIR/WTAP complex elevated m6A levels, thrombospondin 1 (THBS1) expression, and EMT in LECs.</p><p><strong>Conclusions: </strong>LncRNA HOTAIR enhances the assembly of the WTAP/METTL3/METTL14 complex and promotes EMT in LECs by upregulating m6A modification and THBS1 expression. Targeting the HOTAIR/WTAP/THBS1 pathway may prevent or treat PCO.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"20"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12068528/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143967103","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}

{"title":"Spatial and Temporal Changes in Choroid Morphology Associated With Long-Duration Spaceflight.","authors":"Charles Bélanger Nzakimuena, Marissé Masís Solano, Rémy Marcotte-Collard, Mark Richard Lesk, Santiago Costantino","doi":"10.1167/iovs.66.5.17","DOIUrl":"https://doi.org/10.1167/iovs.66.5.17","url":null,"abstract":"<p><strong>Purpose: </strong>Amid efforts to understand spaceflight associated neuro-ocular syndrome (SANS), uncovering the role of the choroid in its etiology is challenged by the accuracy of image segmentation. The present study extended deep learning-based choroid quantification from optical coherence tomography (OCT) to the characterization of pulsatile and topological changes in the macular plane and investigated changes in response to prolonged microgravity exposure.</p><p><strong>Methods: </strong>We analyzed OCT macular videos and volumes acquired from astronauts before, during, and after long-duration spaceflight. Deep learning models were fine-tuned for choroid segmentation and combined with further image processing toward vascularity quantification. Statistical analysis was performed to determine changes in time-dependent and spatially averaged variables from preflight baseline.</p><p><strong>Results: </strong>For 12 astronauts with a mean age of 47 ± 9 years, there were significant increases in choroid thickness and luminal area (LA) averaged over OCT macular video segments. There was also a significant increase in pulsatile LA. For a subgroup of six astronauts for whom inflight imaging was available, choroid volume, luminal volume, and the choroidal vascularity index over the macular region all increased significantly during spaceflight.</p><p><strong>Conclusions: </strong>The findings suggest that localized choroid pulsatile changes occur following prolonged microgravity exposure. They show that the choroid vessels expand in a manner similar to the choroid layer across the macular region during spaceflight, with a relative increase in the space they occupy. The methods developed provide new tools and avenues for studying and establishing effective countermeasures to risks associated with long-duration spaceflight.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"17"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12061066/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144023102","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}

{"title":"Contribution of Leukocidin ED to the Pathogenesis of Staphylococcus aureus Endophthalmitis.","authors":"Luis Longoria-Gonzalez, Phillip S Coburn, Roger Astley, Yan Chen, Michelle C Callegan","doi":"10.1167/iovs.66.5.11","DOIUrl":"https://doi.org/10.1167/iovs.66.5.11","url":null,"abstract":"<p><strong>Purpose: </strong>To test the hypothesis that leukocidin ED (LukED) contributes to the pathogenesis of experimental Staphylococcus aureus endophthalmitis.</p><p><strong>Methods: </strong>Growth curves were generated for S. aureus strain JE2 and strain JE2 lukE::Tn, the transposon mutant of LukED, in brain heart infusion (BHI) and explanted rabbit vitreous. The expression of leukotoxins (lukSF-PV, lukED, hlgABC, and lukGH) was assessed in 18-hour overnight cultures in BHI, tryptic soy broth, and vitreous. S. aureus endophthalmitis was induced by intravitreal injection of 5000 colony-forming units of JE2 or JE2 lukE::Tn into C57BL/6J mice. At 6, 12, and 24 hours after infection, eyes were assessed for retinal function, intraocular colony-forming units and inflammation, and neutrophil infiltration by flow cytometry. RNA was isolated from infected eyes to assess leukotoxin expression.</p><p><strong>Results: </strong>Strains JE2 and JE2 lukE::Tn grew similarly in BHI and vitreous. Transcript levels of leukotoxin subunits were lower in vitreous compared with laboratory media. In vivo, no differences in retinal function, intraocular growth, intraocular inflation, or neutrophil infiltration were observed in eyes infected with JE2 or JE2 lukE::Tn. During infection, other leukotoxins were expressed in vivo in the absence of LukED.</p><p><strong>Conclusions: </strong>LukED does not seem to be essential for the pathogenesis of experimental S. aureus endophthalmitis. However, other leukotoxins are expressed in vivo, which may compensate for the effects of LukED during infection.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"11"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12060071/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143990604","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}

{"title":"Targeting α7 Nicotinic Acetylcholine Receptor for Modulating the Neuroinflammation of Dry Eye Disease Via Macrophages.","authors":"Xujiao Zhou, Yuqing Wu, Yirou Zhang, Binbin Chu, Kan Yang, Jiaxu Hong, Yao He","doi":"10.1167/iovs.66.5.13","DOIUrl":"https://doi.org/10.1167/iovs.66.5.13","url":null,"abstract":"<p><strong>Purpose: </strong>Patients with dry eye disease (DED) often exhibit neurological abnormalities and may even suffer from neuropathic pain and pain-related anxiety or depression. The α-7 nicotinic acetylcholine receptor (α7nAChR) is a pivotal regulator in the anti-inflammatory pathway connecting the nervous and immune systems, Here, we investigate the potential of α7nAChR agonist as a novel treatment for DED.</p><p><strong>Methods: </strong>We induced DED model by unilateral excision of extraorbital lachrymal gland in C57/BL6 mice. After seven days of treatment, RNA sequencing was performed to identify differentially expressed genes in the cornea of lacrimal gland excision (LGE) mice. Quantitative polymerase chain reaction, Western blotting, and flow cytometry tests were carried out to elucidate neuroinflammation changes after α7nAChR activation. Corneal nerve abnormalities were assessed by corneal esthesiometry and immunofluorescence staining.</p><p><strong>Results: </strong>The activation of α7nAChR stimulates genes involved in immune-mediated inflammatory progression and neuroregulation, inhibits the expression of transient receptor potential vanilloid-1, reinstates corneal nerve density, and alleviates anxiety-like behaviors associated with severe DED. Furthermore, we demonstrated that α7nAChR agonist restored corneal nerve abnormality and alleviated inflammation response by down-regulating the proportion of CD86+ M1 macrophages (proinflammatory phenotypes).</p><p><strong>Conclusions: </strong>Our findings underscore the activation of α7nAChR as a pioneering therapeutic approach for preserving corneal nerves balance and controlling inflammation in DED.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"13"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12061059/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143967107","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}

{"title":"Retinoic Acid Receptor Alpha- and Beta-Mediated Signaling Regulates Conjunctival Epithelial Cell Keratinization.","authors":"Hokoru Yoshioka, Mayumi Ueta, Seiichi Yokoo, Norihiko Yokoi, Katsura Mizushima, Yuji Naito, Shigeru Kinoshita, Chie Sotozono","doi":"10.1167/iovs.66.5.6","DOIUrl":"https://doi.org/10.1167/iovs.66.5.6","url":null,"abstract":"<p><strong>Purpose: </strong>Keratinization of the mucosal epithelia develops in severe ocular surface disorders, causing severe visual loss. This study elucidates the molecular mechanisms of keratinization in conjunctival epithelial cells (CjECs) and investigates the involvement of the vitamin A pathway.</p><p><strong>Methods: </strong>Keratinized conjunctival epithelial sheets were generated by a closed system culture of human CjECs and confirmed by immunostaining. Comprehensive gene expression analysis and quantitative real-time PCR (qRT-PCR) were used to examine whether the cells could be used as an in vitro keratinization model. Moreover, immunostaining and qRT-PCR were used to examine alterations of vitamin A pathway-related genes in the cells and also the effect of adding all-trans retinoic acid (ATRA) and retinoic acid receptor alpha/beta (RARA/RARB) agonist Am80. Knockdown of RARA or RARB was also performed using transfection of small interfering RNA to identify receptors for retinoic acid.</p><p><strong>Results: </strong>Immunostaining revealed that CjECs cultured in a closed system had increased expression of keratinization markers. Comprehensive gene expression analysis and qRT-PCR revealed expression changes in vitamin A pathway genes, in addition to keratinization. In the closed system culture, immunostaining revealed that conjunctival epithelial keratinization was suppressed or partially ameliorated by ATRA or Am80, and qRT-PCR revealed that vitamin A pathway-related genes were significantly altered. Moreover, knockdown of RARA or RARB induced an increase in keratinization marker involucrin.</p><p><strong>Conclusions: </strong>Keratinization of CjECs involves RARA or RARB-mediated pathways, and ATRA and Am80 alter the expression of vitamin A pathway gene and suppress keratinization.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 5","pages":"6"},"PeriodicalIF":5.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12054659/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144025045","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}