The Role of LOX-1 in Innate Immunity to Aspergillus fumigatus in Corneal Epithelial Cells
The Role of LOX-1 in Innate Immunity to Aspergillus fumigatus in Corneal Epithelial Cells
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LOX-1 在角膜上皮细胞对烟曲霉先天免疫中的作用。
DOI：
10.1167/iovs.14-15989
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发表时间：
2015-06-01
影响因子：
4.4
通讯作者：
Liu, Ying
中科院分区：
文献类型：
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作者：
Li, Cui;Zhao, Guiqiu;Liu, Ying
目的.探讨凝集素样氧化低密度脂蛋白受体1（LOX-1）在真菌感染角膜上皮细胞中的作用。真菌性角膜炎角膜13只为实验组。10例健康供体角膜为对照组。刮取角膜上皮并检测LOX-1 mRNA。免疫组化法检测LOX-1蛋白的表达。用75%乙醇灭活的烟曲霉处理人角膜上皮细胞（HCECs）。检测LOX-1，并检测CXCL 1、TNF-α、IL-6和dectin-1水平，无论是否中和LOX-1。建立大鼠真菌性角膜炎模型，采用PCR、免疫组化和Western blot检测LOX-1的表达。在LOX-1中和前后检测磷酸化p38 MAPK（p-p38 MAPK）蛋白; CXCL 1、TNF-α和IL-6 mRNA水平;和髓过氧化物酶（MPO）。LOX-1表达于角膜上皮。体外细胞实验显示，正常人HCECs中有LOX-1表达，A. LOX-1蛋白表达在刺激后24和48小时增加。LOX-1的中和降低了CXCL 1、TNF-α的表达，但没有改变IL-6或dectin-1的表达。在大鼠中发生的烟曲霉角膜炎激活p38 MAPK，并通过LOX-1升高CXCL 1、TNF-α和IL-6的表达。LOX-1中和降低MPO水平。LOX-1在正常角膜上皮、HCECs和A.烟曲霉通过LOX-1上调CXCL 1和TNF-α的表达。大鼠A.烟曲霉角膜炎激活p38 MAPK并通过LOX-1升高CXCL 1、TNF-α和IL-6的表达。
PURPOSE. To determine the role of lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) in corneal epithelial cells exposed to fungus.METHODS. Thirteen corneas with fungal keratitis composed the experimental group. Ten healthy donor corneas were the control group. Corneal epithelium was scraped and LOX-1 mRNA tested. Immunostaining was used to detect LOX-1 protein. Human corneal epithelial cells (HCECs) were treated with 75% ethanol-killed Aspergillus fumigatus. LOX-1 was detected, and CXCL1, TNF-alpha, IL-6, and dectin-1 levels were detected with or without neutralization of LOX-1. Rat fungal keratitis was establi7shed and LOX-1 was detected by PCR, immunostaining, and Western blot. Phosphorylated-p38MAPK (p-p38MAPK) protein; CXCL1, TNF-alpha, and IL-6 mRNA levels; and myeloperoxidase (MPO) were tested before and after LOX-1 neutralization.RESULTS. LOX-1 was expressed in corneal epithelium. In vitro cellular experiment showed that LOX-1 was detected in normal HCECs, and LOX-1 mRNA increased after stimulation of A. fumigatus and peaked at 8 hours; LOX-1 protein expression increased after stimulation at 24 and 48 hours. Neutralization of LOX-1 decreased expression of CXCL1, TNF-alpha, but did not change IL-6 or dectin-1 expression. Aspergillus fumigatus keratitis developed in rats activated p38MAPK and elevated the expression of CXCL1, TNF-alpha, and IL-6 through LOX-1. LOX-1 neutralization reduced MPO levels.CONCLUSIONS. LOX-1 expressed in normal corneal epithelium and HCECs and A. fumigatus elevated the expression of CXCL1 and TNF-alpha through LOX-1. Rat A. fumigatus keratitis activated p38MAPK and elevated the expression of CXCL1, TNF-alpha, and IL-6 through LOX-1.