Homocysteine Induces Brain and Retinal Microvascular Endothelial Cell Barrier Damage and Hyperpermeability via NLRP3 Inflammasome Pathway Differentially

IF 1.9 4区医学 Q3 HEMATOLOGY

Microcirculation Pub Date : 2025-07-17 DOI:10.1111/micc.70019

O'lisa Yaa Waithe, Aliyah Anderson, Saravanakumar Muthusamy, Gabriela M. Seplovich, Binu Tharakan

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引用次数: 0

Abstract

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.

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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

氨基酸同型半胱氨酸（HCY）与几种疾病的病理生物学有关，包括航天相关的神经-眼综合征（SANS）——一种影响宇航员近视力的症状集合。血视网膜屏障（BRB）和血脑屏障（BBB）功能障碍与SANS的病理生物学有关。我们的目的是评估HCY如何影响BRB/BBB的通透性，以及NLRP3炎症小体在调节这种影响中的作用。方法分别用100 μM HCY或联合1 μM NLRP3炎性体抑制剂MCC950对人脑和视网膜微血管内皮细胞（HBMECs和HRMECs）进行处理。采用荧光法测定细胞活力，caspase-1酶测定，RT-PCR法表达BRB/BBB紧密连接蛋白ZO-1 (zonula occluden -1)， fitc -葡聚糖法检测屏障通透性。结果在hrmec和hbmec中，hcy诱导的内皮单层高通透性显著（p < 0.05）。在HBMECs中，MCC950的作用减弱（p < 0.05）。在添加HCY后，两种细胞类型均观察到Caspase-1活性增加。添加HCY后，观察到屏障受损的基因表达结果，特别是ZO-1 （p < 0.05）。结论HCY诱导视网膜和脑内皮细胞高通透性。与视网膜内皮细胞相比，nlrp3介导的hcy诱导的微血管通透性在脑内皮细胞中更为突出。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microcirculation 医学-外周血管病

CiteScore

5.00

自引率

4.20%

发文量

审稿时长

6-12 weeks

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.