紧密连接蛋白 ZO-1 对实验性角膜新生血管形成的抑制作用

IF 1.9 4区 医学 Q2 OPHTHALMOLOGY Journal of Ocular Pharmacology and Therapeutics Pub Date : 2024-07-01 DOI:10.1089/jop.2023.0162
Qingying Yao, Hongya Wu, Hang Ren, Jiufa Cao, Ying Shao, Gaoqin Liu, Peirong Lu
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引用次数: 0

摘要

目的:探讨紧密连接蛋白Zonula occludens 1(ZO-1)对实验性角膜新生血管(CNV)的影响。方法:用黑白猫科动物左眼建立 CNV 模型:用 NaOH 在 BALB/c 小鼠左眼建立 CNV 模型。造模后在烧伤的角膜上局部涂抹抗 ZO-1 中和抗体,每天三次,持续一周。使用角膜全层荧光免疫组化检测法分析 CD31 的表达,计算 CNV 数量与面积之比。ZO-1、血管内皮生长因子(VEGF)、白细胞介素(IL)-1β、IL-6、IL-8、IL-18、单核细胞趋化蛋白-1(MCP-1)的信使核糖核酸(mRNA)和蛋白表达水平、通过逆转录酶聚合酶链反应(PCR)和免疫印迹分析检测了烧伤角膜中的肿瘤坏死因子α(TNF-α)、磷酸化蛋白激酶 C(pPKC)和集束蛋白。流式细胞术检测了中性粒细胞、巨噬细胞和祖细胞的浸润情况。结果显示45 秒碱损伤组的 CNV 明显高于 15 秒碱损伤组。在另一项实验中,ZO-1 抗体组的 CNV 明显高于药物治疗组。ZO-1 抗体组角膜血管内皮生长因子、IL-1β、IL-6、IL-8、IL-18 和 MCP-1 的 mRNA 和蛋白表达水平明显高于对照组。中性粒细胞、巨噬细胞和祖细胞的浸润在 ZO-1 抗体组明显高于对照组。45 秒碱损伤组的 TNF-α 表达量远高于 15 秒碱损伤组。然而,pPKC 和 clusterin 蛋白表达在 45 秒碱损伤组远低于 15 秒碱损伤组。结论抗ZO-1中和抗体处理的小鼠通过增强角膜内祖细胞和炎症细胞的浸润,表现出更强的碱诱导的CNV。
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Inhibition of Experimental Corneal Neovascularization by the Tight Junction Protein ZO-1.

Purpose: To explore the effects of the tight junction protein zonula occludens 1 (ZO-1) on experimental corneal neovascularization (CNV). Methods: CNV models were established in the left eyes of BALB/c mice using NaOH. Anti-ZO-1 neutralizing antibody was topically applied to the burnt corneas after modeling thrice a day for 1 week. CD31 expression was analyzed to calculate the ratio of CNV number to area using a corneal whole-mount fluorescent immunohistochemical assay. Messenger ribonucleic acid (mRNA) and protein expression levels of ZO-1, vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-6, IL-8, IL-18, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor alpha (TNF-α), phosphorylated protein kinase C (pPKC), and clusterin in burned corneas were detected by reverse transcriptase polymerase chain reaction (PCR) and western blot analyses. Infiltration of neutrophils, macrophages, and progenitor cells was examined by flow cytometry. Results: CNV was obviously greater in 45 s than in 15 s alkali injury group. In another experiment, CNV was obviously greater in the ZO-1 antibody group than in the vehicle-treated group. Corneal mRNA and protein expression levels of VEGF, IL-1β, IL-6, IL-8, IL-18, and MCP-1 were significantly higher in the ZO-1 antibody group than in the control group. Infiltration of neutrophils, macrophages, and progenitor cells was significantly greater in the ZO-1 antibody group than in the control group. TNF-α expression was much higher in 45 s than in 15 s alkali injury group. However, protein expression of pPKC and clusterin was much lower in 45 s than in 15 s alkali injury group. Conclusions: Anti-ZO-1 neutralizing antibody-treated mice exhibited enhanced alkali-induced CNV through enhanced intracorneal infiltration of progenitor and inflammatory cells.

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来源期刊
CiteScore
4.60
自引率
4.30%
发文量
72
审稿时长
1 months
期刊介绍: Journal of Ocular Pharmacology and Therapeutics is the only peer-reviewed journal that combines the fields of ophthalmology and pharmacology to enable optimal treatment and prevention of ocular diseases and disorders. The Journal delivers the latest discoveries in the pharmacokinetics and pharmacodynamics of therapeutics for the treatment of ophthalmic disorders. Journal of Ocular Pharmacology and Therapeutics coverage includes: Glaucoma Cataracts Retinal degeneration Ocular infection, trauma, and toxicology Ocular drug delivery and biotransformation Ocular pharmacotherapy/clinical trials Ocular inflammatory and immune disorders Gene and cell-based therapies Ocular metabolic disorders Ocular ischemia and blood flow Proliferative disorders of the eye Eyes on Drug Discovery - written by Gary D. Novack, PhD, featuring the latest updates on drug and device pipeline developments as well as policy/regulatory changes by the FDA.
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