Schicht Martin, Rausch Felix, Beron Martin, Jacobi Christina, Garreis Fabian, Hartjen Nadine, Beileke Stephanie, Kruse Friedrich, Bräuer Lars, Paulsen Friedrich
Department of Anatomy II Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
Department of Ophthalmology, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
Invest Ophthalmol Vis Sci. 2015 Nov;56(12):7312-23. doi: 10.1167/iovs.15-17560.
Palate Lung Nasal Clone (PLUNC) is a hydrophobic protein belonging to the family of surfactant proteins that is involved in fluid balance regulation of the lung. Moreover, it is known to directly act against gram-negative bacteria. The purpose of this study was to investigate the possible expression and antimicrobial role of PLUNC at the healthy ocular surface and in tears of patients suffering from dry eye disease (DED).
Bioinformatics and biochemical and immunologic methods were combined to elucidate the structure and function of PLUNC at the ocular surface. Tissue-specific localization was performed by using immunohistochemistry. The PLUNC levels in tear samples from non-Sjögren's DED patients with moderate dry eye suffering either from hyperevaporation or tear deficiency were analyzed by ELISA and compared with tears from healthy volunteers.
Palate Lung Nasal Clone is expressed under healthy conditions at the ocular surface and secreted into the tear film. Protein modeling studies and molecular dynamics simulations performed indicated surface activity of PLUNC. In vitro experiments revealed that proinflammatory cytokines and bacterial supernatants have only a slight effect on the expression of PLUNC in HCE and HCjE cell lines. In tears from DED patients, the PLUNC concentration is significantly increased (7-fold in evaporative dry eye tears and 17-fold in tears from patients with tear deficiency) compared with healthy subjects.
The results show that PLUNC is a protein of the tear film and suggest that it plays a role in fluid balance and surface tension regulation at the ocular surface.
腭肺鼻克隆蛋白(PLUNC)是一种疏水蛋白,属于表面活性蛋白家族,参与肺的液体平衡调节。此外,已知它能直接对抗革兰氏阴性菌。本研究的目的是调查PLUNC在健康眼表及干眼疾病(DED)患者泪液中的可能表达及抗菌作用。
结合生物信息学、生化和免疫学方法阐明PLUNC在眼表的结构和功能。采用免疫组织化学进行组织特异性定位。通过酶联免疫吸附测定(ELISA)分析非干燥综合征的中度干眼患者(因蒸发过强或泪液缺乏所致)泪液样本中的PLUNC水平,并与健康志愿者的泪液进行比较。
腭肺鼻克隆蛋白在健康条件下于眼表表达并分泌至泪膜中。进行的蛋白质建模研究和分子动力学模拟表明PLUNC具有表面活性。体外实验显示,促炎细胞因子和细菌上清液对人角膜上皮(HCE)和人结膜上皮(HCjE)细胞系中PLUNC的表达仅有轻微影响。与健康受试者相比,DED患者泪液中的PLUNC浓度显著升高(蒸发过强型干眼泪液中升高7倍,泪液缺乏型患者泪液中升高17倍)。
结果表明PLUNC是泪膜中的一种蛋白,并提示其在眼表液体平衡和表面张力调节中发挥作用。