Wang Zhe, Chen Ziyan, Yang Qian, Jiang Yibo, Lin Liping, Liu Xialin, Wu Kaili
Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, China.
Invest Ophthalmol Vis Sci. 2014 Sep 25;55(10):6722-7. doi: 10.1167/iovs.14-14491.
The aim of this study was to explore the possible interactions between vitelline membrane outer layer 1 homolog (VMO1) and other tear proteins and to determine the function of VMO1 in tear fluid.
Interactions between recombinant human VMO1 and several abundant tear proteins were determined by dot blot, His pull-down, immunoprecipitation, and Western blot assays, as well as by computer-assisted prediction and modeling of molecular interactions. Kirby-Bauer antibiotic testing was performed to determine whether VMO1 possesses antimicrobial activity. Tear samples were collected from dry eye patients and from healthy controls. The role of VMO1 in maintaining the stability of tear film was investigated by measurement of contact angles on Teflon, tear break-up time (TBUT) and the time-dependent reduction in tear film integrity in mice.
Vitelline membrane outer layer 1 homolog showed an interaction with lysozyme C (LYSC) in the dot-blot, His pull-down, and immunoprecipitation assays. Vitelline membrane outer layer 1 homolog revealed no zones of growth inhibition of standard strains of Staphylococcus aureus and Escherichia coli. Tears presented smaller contact angles on Teflon surfaces after the addition of VMO1 (P<0.05). Vitelline membrane outer layer 1 homolog-treated mice showed longer TBUTs (P<0.05). Tear films from VMO1-treated mice maintained their integrity for longer periods of time than tear films from the control group, and this effect was dose-dependent.
Vitelline membrane outer layer 1 homolog interacts with LYSC and has positive effects on the stabilization of tear film.
本研究旨在探讨卵黄膜外层1同源物（VMO1）与其他泪液蛋白之间可能的相互作用，并确定VMO1在泪液中的功能。
通过斑点印迹、His下拉、免疫沉淀和蛋白质印迹分析，以及计算机辅助的分子相互作用预测和建模，确定重组人VMO1与几种丰富的泪液蛋白之间的相互作用。进行Kirby-Bauer抗生素测试以确定VMO1是否具有抗菌活性。从干眼患者和健康对照者中收集泪液样本。通过测量特氟龙上的接触角、泪膜破裂时间（TBUT）以及小鼠泪膜完整性随时间的降低情况，研究VMO1在维持泪膜稳定性中的作用。
在斑点印迹、His下拉和免疫沉淀分析中，卵黄膜外层1同源物显示与溶菌酶C（LYSC）有相互作用。卵黄膜外层1同源物对金黄色葡萄球菌和大肠杆菌的标准菌株未显示生长抑制区域。添加VMO1后，泪液在特氟龙表面呈现较小的接触角（P<0.05）。经卵黄膜外层1同源物处理的小鼠显示出更长的TBUTs（P<0.05）。与对照组相比，经VMO1处理的小鼠的泪膜在更长时间内保持其完整性，且这种作用呈剂量依赖性。
卵黄膜外层1同源物与LYSC相互作用，并对泪膜的稳定有积极作用。
