Liu Gaoqin, Wu Hongya, Lu Peirong, Zhang Xueguang
a Department of Ophthalmology , The First Affiliated Hospital of Soochow University , Suzhou , P.R. China.
b Jiangsu Clinical Immunology Institute, The First Affiliated Hospital of Soochow University , Suzhou , P.R. China.
Curr Eye Res. 2017 Mar;42(3):368-379. doi: 10.1080/02713683.2016.1196705. Epub 2016 Jul 15.
To explore the roles of interleukin (IL)-17A in alkali-induced corneal neovascularization (CRNV).
Induction of CRNV by alkali injury was performed and compared in recombinant mouse IL-17A protein or anti-mouse IL-17A neutralizing antibody (Ab)-treated mice. The expression of IL-17A, IL-17A receptor (IL-17RA), vascular endothelial growth factor (VEGF), IL-6, IL-8, and monocyte chemoattractant protein-1 (MCP-1) in burned corneas or other cells was examined by RT-PCR and western blot. The infiltration of macrophages, neutrophils, and c-Kit-positive progenitor cells into burned corneas was detected by flow cytometry (FCM). The Raw264.7 cell line and mouse corneal fibroblasts were stimulated by mouse recombinant IL-17A or anti-mouse IL-17A neutralizing Ab in vitro, and the expression of IL-6 and VEGF in culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA).
The alkali-induced CRNV peaked 2 weeks after the alkali burn. Compared with the vehicle-treated group, mouse recombinant IL-17A administration significantly increased the amount of CRNV, and it decreased in mice treated with neutralizing anti-mouse IL-17A Ab. RT-PCR and western blot confirmed that the IL-17A upregulated intracorneal VEGF, IL-6, IL-8, MCP-1, and phosphorylated focal adhesion kinase (pFAK) expression. FCM revealed that the infiltration of intracorneal progenitor cells, M1 macrophages, and neutrophils was also augmented in IL-17A-treated mice. The ELISA showed that IL-17A markedly induced VEGF and IL-6 expression in the Raw264.7 murine macrophage cell line and in the mouse corneal fibroblasts.
IL-17A-treated mice exhibited enhanced alkali-induced CRNV through enhanced intracorneal progenitor cell and inflammatory cell infiltration, and increased VEGF and IL-6 expression by fibroblasts and macrophages.
探讨白细胞介素（IL）-17A在碱诱导的角膜新生血管化（CRNV）中的作用。
通过碱烧伤诱导CRNV，并在重组小鼠IL-17A蛋白或抗小鼠IL-17A中和抗体（Ab）处理的小鼠中进行比较。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测烧伤角膜或其他细胞中IL-17A、IL-17A受体（IL-17RA）、血管内皮生长因子（VEGF）、IL-6、IL-8和单核细胞趋化蛋白-1（MCP-1）的表达。通过流式细胞术（FCM）检测巨噬细胞、中性粒细胞和c-Kit阳性祖细胞向烧伤角膜的浸润情况。在体外，用小鼠重组IL-17A或抗小鼠IL-17A中和Ab刺激Raw264.7细胞系和小鼠角膜成纤维细胞，通过酶联免疫吸附测定（ELISA）法测定培养上清液中IL-6和VEGF的表达。
碱诱导的CRNV在碱烧伤后2周达到峰值。与载体处理组相比，给予小鼠重组IL-17A显著增加了CRNV的数量，而在用中和抗小鼠IL-17A Ab处理的小鼠中CRNV数量减少。RT-PCR和蛋白质免疫印迹法证实，IL-17A上调了角膜内VEGF、IL-6、IL-8、MCP-1和磷酸化粘着斑激酶（pFAK）的表达。FCM显示，在IL-17A处理的小鼠中，角膜内祖细胞、M1巨噬细胞和中性粒细胞的浸润也增加。ELISA显示，IL-17A显著诱导Raw264.7小鼠巨噬细胞系和小鼠角膜成纤维细胞中VEGF和IL-6的表达。
IL-17A处理的小鼠通过增强角膜内祖细胞和炎性细胞浸润，以及成纤维细胞和巨噬细胞增加VEGF和IL-6表达，表现出增强的碱诱导CRNV。
