{"title":"同型半胱氨酸通过NLRP3炎性小体通路诱导脑和视网膜微血管内皮细胞屏障损伤和高通透性","authors":"O'lisa Yaa Waithe, Aliyah Anderson, Saravanakumar Muthusamy, Gabriela M. Seplovich, Binu Tharakan","doi":"10.1111/micc.70019","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Objectives</h3>\n \n <p>The amino acid homocysteine (HCY) has been implicated in the pathobiology of several conditions, including spaceflight-associated neuro-ocular syndrome (SANS)—a collection of symptoms affecting near vision in astronauts. Blood-retinal barrier (BRB) and blood–brain barrier (BBB) dysfunctions are implicated in the pathobiology of SANS. Our objective was to assess how HCY affects BRB/BBB permeability and the role of the NLRP3 inflammasome in the modulation of such effects.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Human brain and retinal microvascular endothelial cells (HBMECs and HRMECs) were treated with 100 μM HCY alone or in conjunction with NLRP3 inflammasome inhibitor MCC950 at 1 μM. The assays performed included fluorometric assays to measure cell viability, an enzyme assay for caspase-1, expression of BRB/BBB tight junction protein zonula occludens-1 (ZO-1) by RT-PCR, and barrier permeability using FITC-dextran.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>In HRMECs and HBMECs, HCY-induced endothelial monolayer hyperpermeability significantly (<i>p</i> < 0.05). In HBMECs, the effect was attenuated by MCC950 (<i>p</i> < 0.05). Increased Caspase-1 activity was observed in both cell types following the addition of HCY. Following HCY addition, gene expression results denoting barrier damage were observed, particularly that of ZO-1 (<i>p</i> < 0.05).</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>HCY induces hyperpermeability in retinal and brain endothelial cells. NLRP3-mediation in HCY-induced microvascular permeability is prominent in brain endothelial cells compared to retinal endothelial cells.</p>\n </section>\n </div>","PeriodicalId":18459,"journal":{"name":"Microcirculation","volume":"32 5","pages":""},"PeriodicalIF":1.9000,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Homocysteine Induces Brain and Retinal Microvascular Endothelial Cell Barrier Damage and Hyperpermeability via NLRP3 Inflammasome Pathway Differentially\",\"authors\":\"O'lisa Yaa Waithe, Aliyah Anderson, Saravanakumar Muthusamy, Gabriela M. Seplovich, Binu Tharakan\",\"doi\":\"10.1111/micc.70019\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Objectives</h3>\\n \\n <p>The amino acid homocysteine (HCY) has been implicated in the pathobiology of several conditions, including spaceflight-associated neuro-ocular syndrome (SANS)—a collection of symptoms affecting near vision in astronauts. Blood-retinal barrier (BRB) and blood–brain barrier (BBB) dysfunctions are implicated in the pathobiology of SANS. Our objective was to assess how HCY affects BRB/BBB permeability and the role of the NLRP3 inflammasome in the modulation of such effects.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>Human brain and retinal microvascular endothelial cells (HBMECs and HRMECs) were treated with 100 μM HCY alone or in conjunction with NLRP3 inflammasome inhibitor MCC950 at 1 μM. The assays performed included fluorometric assays to measure cell viability, an enzyme assay for caspase-1, expression of BRB/BBB tight junction protein zonula occludens-1 (ZO-1) by RT-PCR, and barrier permeability using FITC-dextran.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>In HRMECs and HBMECs, HCY-induced endothelial monolayer hyperpermeability significantly (<i>p</i> < 0.05). In HBMECs, the effect was attenuated by MCC950 (<i>p</i> < 0.05). Increased Caspase-1 activity was observed in both cell types following the addition of HCY. Following HCY addition, gene expression results denoting barrier damage were observed, particularly that of ZO-1 (<i>p</i> < 0.05).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusions</h3>\\n \\n <p>HCY induces hyperpermeability in retinal and brain endothelial cells. NLRP3-mediation in HCY-induced microvascular permeability is prominent in brain endothelial cells compared to retinal endothelial cells.</p>\\n </section>\\n </div>\",\"PeriodicalId\":18459,\"journal\":{\"name\":\"Microcirculation\",\"volume\":\"32 5\",\"pages\":\"\"},\"PeriodicalIF\":1.9000,\"publicationDate\":\"2025-07-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microcirculation\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/micc.70019\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"HEMATOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microcirculation","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/micc.70019","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"HEMATOLOGY","Score":null,"Total":0}
Homocysteine Induces Brain and Retinal Microvascular Endothelial Cell Barrier Damage and Hyperpermeability via NLRP3 Inflammasome Pathway Differentially
Objectives
The amino acid homocysteine (HCY) has been implicated in the pathobiology of several conditions, including spaceflight-associated neuro-ocular syndrome (SANS)—a collection of symptoms affecting near vision in astronauts. Blood-retinal barrier (BRB) and blood–brain barrier (BBB) dysfunctions are implicated in the pathobiology of SANS. Our objective was to assess how HCY affects BRB/BBB permeability and the role of the NLRP3 inflammasome in the modulation of such effects.
Methods
Human brain and retinal microvascular endothelial cells (HBMECs and HRMECs) were treated with 100 μM HCY alone or in conjunction with NLRP3 inflammasome inhibitor MCC950 at 1 μM. The assays performed included fluorometric assays to measure cell viability, an enzyme assay for caspase-1, expression of BRB/BBB tight junction protein zonula occludens-1 (ZO-1) by RT-PCR, and barrier permeability using FITC-dextran.
Results
In HRMECs and HBMECs, HCY-induced endothelial monolayer hyperpermeability significantly (p < 0.05). In HBMECs, the effect was attenuated by MCC950 (p < 0.05). Increased Caspase-1 activity was observed in both cell types following the addition of HCY. Following HCY addition, gene expression results denoting barrier damage were observed, particularly that of ZO-1 (p < 0.05).
Conclusions
HCY induces hyperpermeability in retinal and brain endothelial cells. NLRP3-mediation in HCY-induced microvascular permeability is prominent in brain endothelial cells compared to retinal endothelial cells.
期刊介绍：
The journal features original contributions that are the result of investigations contributing significant new information relating to the vascular and lymphatic microcirculation addressed at the intact animal, organ, cellular, or molecular level. Papers describe applications of the methods of physiology, biophysics, bioengineering, genetics, cell biology, biochemistry, and molecular biology to problems in microcirculation.
Microcirculation also publishes state-of-the-art reviews that address frontier areas or new advances in technology in the fields of microcirculatory disease and function. Specific areas of interest include: Angiogenesis, growth and remodeling; Transport and exchange of gasses and solutes; Rheology and biorheology; Endothelial cell biology and metabolism; Interactions between endothelium, smooth muscle, parenchymal cells, leukocytes and platelets; Regulation of vasomotor tone; and Microvascular structures, imaging and morphometry. Papers also describe innovations in experimental techniques and instrumentation for studying all aspects of microcirculatory structure and function.
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