Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China.
Invest Ophthalmol Vis Sci. 2018 Jan 1;59(1):166-175. doi: 10.1167/iovs.17-21887.
To determine the role of protease-activated receptor-2 (PAR-2) in cornea infected by Aspergillus fumigatus.
PAR-2 was tested in normal and infected corneas of C57BL/6 mice. Mice corneas were infected with A. fumigatus with or without pretreatment of PAR-2 antagonist (FSLLRY-NH2). Polymorphonuclear neutrophilic leukocytes (PMNs) were stimulated with 75% ethanol-killed A. fumigatus with or without pretreatment of FSLLRY-NH2. Disease severity was documented by clinical score and photographs with a slit lamp. PCR, Western blot, and ELISA tested expression of PAR-2, IL-1β, TNF-α, IFN-γ, MIP-2, and p-ERK1/2. PMN infiltration was assessed by myeloperoxidase assay and immunofluorescent staining.
PAR-2 expression was significantly elevated by A. fumigatus, whereas the upregulation was significantly inhibited by FSLLRY-NH2 in mice corneas. FSLLRY-NH2 decreased disease response, PMN infiltration, and proinflammatory cytokine expression compared with infected control. In PMNs, PAR-2 expression was also significantly increased by A. fumigatus, which was significantly inhibited by FSLLRY-NH2. FSLLRY-NH2 significantly inhibited proinflammatory cytokine protein expression, as compared with that in infected control cells, which may be modified by p-ERK1/2.
These data provide evidence that A. fumigatus increased PAR-2 expression and elevated disease, PMN infiltration, and proinflammatory cytokine expression through PAR-2, which may be modified by p-ERK1/2.
确定蛋白酶激活受体 2（PAR-2）在感染烟曲霉菌的角膜中的作用。
检测 C57BL/6 小鼠正常和感染角膜中的 PAR-2。用烟曲霉菌感染小鼠角膜，并用或不用 PAR-2 拮抗剂（FSLLRY-NH2）预处理。用 75%乙醇杀死的烟曲霉菌刺激多形核中性粒细胞（PMN），并用或不用 FSLLRY-NH2 预处理。用临床评分和裂隙灯照片记录疾病严重程度。PCR、Western blot 和 ELISA 检测 PAR-2、IL-1β、TNF-α、IFN-γ、MIP-2 和 p-ERK1/2 的表达。用髓过氧化物酶测定和免疫荧光染色评估 PMN 浸润。
烟曲霉菌显著上调 PAR-2 的表达，而 FSLLRY-NH2 可显著抑制小鼠角膜中的上调。与感染对照组相比，FSLLRY-NH2 降低了疾病反应、PMN 浸润和促炎细胞因子的表达。在 PMN 中，烟曲霉菌也显著增加了 PAR-2 的表达，而 FSLLRY-NH2 则显著抑制了其表达。与感染对照组细胞相比，FSLLRY-NH2 显著抑制了促炎细胞因子蛋白的表达，这可能通过 p-ERK1/2 进行修饰。
这些数据提供的证据表明，烟曲霉菌通过 PAR-2 增加 PAR-2 的表达，从而导致疾病、PMN 浸润和促炎细胞因子的表达升高，这可能通过 p-ERK1/2 进行修饰。
