Abstract
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.
Highlights
Aqueous humor (AH) is a transparent liquid secreted by the ciliary epithelium
21 cataract AH samples were used to generate a comprehensive profile of the human AH proteome
The functional annotation was provided by the Gene Ontology (GO) annotation and Ingenuity Pathway Analysis (IPA) analysis
Summary
Aqueous humor (AH) is a transparent liquid secreted by the ciliary epithelium. It is mainly composed of 99.9% water, and trace amounts of sugar, vitamins, protein, and other factors (Macknight et al, 2000). The functions of AH are to maintain intraocular pressure, nourish the avascular cornea and lens, and remove metabolic waste (Bill, 1975). It plays an important role in the pathogenesis and progression of ophthalmic disease (Kliuchnikova et al, 2016). The analysis of the proteome in AH is important to understand the physiology and changes evoked by pathological situations, especially in posterior disorders
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