Yu Mengxi, Xie Feng, Liu Xiang, Sun Haidan, Guo Zhengguang, Liu Xiaoyan, Li Wei, Sun Wei, Wang Ying, He Chengyan
China-Japan Union Hospital of Jilin University, Changchun, China.
Shanghai AB Sciex Analytical Instrument Trading Co., Ltd., Shanghai, China.
Front Mol Biosci. 2020 Oct 8;7:587677. doi: 10.3389/fmolb.2020.587677. eCollection 2020.
Aqueous humor (AH) proteins are involved in many physiological and pathological processes of the eye. The proteome analysis of AH is important to understand its physiological and pathophysiological functions. In the present study, AH samples obtained from 21 cataract volunteers were pooled together. After high-pH RPLC offline separation, the pooled sample was analyzed by LC-MS/MS to provide a comprehensive profile of AH proteome. The function analysis was provided by the GO and IPA annotation. In order to determine whether the AH proteome can reflect the pathophysiological changes of the disease, DIA technology was used to analyze the AH samples obtained from three neovascular glaucoma (NVG) patients (six samples) before and after drug treatment. The differential proteins were validated by PRM technology in an independent group (14 samples). In the AH proteome database, 802 proteins were identified, and 318 proteins were identified for the first time. Furthermore, 480 proteins were quantified based on the peak intensity-based semiquantification (iBAQ), which ranged by approximately 7 orders of magnitude. These proteins are primarily involved in immunity- and inflammation-related pathways. The differential AH proteomic analysis in NVG treatment revealed that the AH proteome can reflect the pathophysiological changes of drug treatment. Angiogenesis and thrombus coagulation progression are deeply involved in NVG treatment. The present experiment provided a comprehensive AH proteome analysis and expanded the profile of human AH proteome. The differential AH proteomic analysis of NVG treatment indicated that AH proteome can reflect the pathophysiological changes in drug intervention.
房水(AH)蛋白参与眼睛的许多生理和病理过程。房水的蛋白质组分析对于理解其生理和病理生理功能很重要。在本研究中,将从21名白内障志愿者获得的房水样本汇集在一起。经过高pH值反相液相色谱离线分离后,对汇集样本进行液相色谱-串联质谱分析,以提供房水蛋白质组的全面概况。通过基因本体(GO)和Ingenuity Pathway Analysis(IPA)注释进行功能分析。为了确定房水蛋白质组是否能反映疾病的病理生理变化,采用数据非依赖采集(DIA)技术分析了3例新生血管性青光眼(NVG)患者药物治疗前后的房水样本(6个样本)。差异蛋白在一个独立组(14个样本)中通过平行反应监测(PRM)技术进行验证。在房水蛋白质组数据库中,鉴定出802种蛋白质,其中318种蛋白质为首次鉴定。此外,基于峰强度的半定量(iBAQ)对480种蛋白质进行了定量,其范围约为7个数量级。这些蛋白质主要参与免疫和炎症相关途径。NVG治疗中的差异房水蛋白质组分析表明,房水蛋白质组可以反映药物治疗的病理生理变化。血管生成和血栓形成进程在NVG治疗中密切相关。本实验提供了全面的房水蛋白质组分析,并扩展了人类房水蛋白质组的概况。NVG治疗的差异房水蛋白质组分析表明,房水蛋白质组可以反映药物干预中的病理生理变化。