目的：探讨紧密连接蛋白闭塞带1（ZO-1）对实验性角膜新生血管（CNV）的影响。方法：采用NaOH建立BALB/c小鼠左眼CNV模型。建模后每天三次，持续 1 周，将抗 ZO-1 中和抗体局部涂抹在烧伤的角膜上。使用角膜整体荧光免疫组织化学测定分析 CD31 表达以计算 CNV 数量与面积的比率。 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-) 的信使核糖核酸 (mRNA) 和蛋白表达水平1)、通过逆转录聚合酶链反应 (PCR) 和蛋白质印迹分析检测烧伤角膜中的肿瘤坏死因子 α (TNF-α)、磷酸化蛋白激酶 C (pPKC) 和簇蛋白。通过流式细胞术检查中性粒细胞、巨噬细胞和祖细胞的浸润。结果：45 s碱伤组CNV明显大于15 s碱伤组。在另一项实验中，ZO-1抗体组的CNV明显高于媒介物处理组。 ZO-1抗体组角膜VEGF、IL-1β、IL-6、IL-8、IL-18和MCP-1 mRNA和蛋白表达水平显着高于对照组。 ZO-1抗体组的中性粒细胞、巨噬细胞和祖细胞的浸润显着高于对照组。 45 s碱损伤组TNF-α表达明显高于15 s碱损伤组。然而，45 s碱损伤组pPKC和clusterin蛋白表达明显低于15 s碱损伤组。结论：抗 ZO-1 中和抗体治疗的小鼠通过增强祖细胞和炎症细胞的角膜内浸润而表现出增强的碱诱导的 CNV。

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.