Acute angle-closure glaucoma (AACG) is an ophthalmic emergency that occurs over the course of hours or days and may cause irreversible blindness if not treated immediately. In most cases, optic nerve damage is the cause of visual field (VF) loss in AACG. There has been no reliable biomarker found to evaluate optic nerve damage to date. Aqueous humor (AH) proteome analysis might reveal the proteomic alterations in AACG and provide helpful clues in the search for an AH biomarker of optic nerve damage and VF loss.In this study, we used the AH proteome to explore the functions of differentially expressed proteins (DEPs) during disease progression. The AH proteins from the early-stage group and late-stage group were extracted and analyzed by the data-independent acquisition (DIA) method. The DEPs functions were annotated, and parallel reaction monitoring (PRM) was used to validate the key DEPs.A total of 87 DEPs were found. Gene Ontology analysis showed that most DEPs were enriched in immunology, hemodynamics, and apoptosis. Ingenuity pathway analysis found that vascular endothelial growth factor (VEGF) signaling, the production of reactive oxygen species (ROS) in macrophages, and the nuclear factor erythroid 2-related factor 2 (NRF2)-mediated oxidative stress response were active pathways in the late stage of AACG. The mechanism of retinal ganglion cell (RGC) death was hypothesized on the basis of DEP functional analysis. A total of 20 DEPs were validated by using PRM, and prostaglandin-H2 D-isomerase was found to have the potential to evaluate optic nerve damage.This study showed that AH proteomic analysis could reveal the proteomic alterations in the pathogenesis of VF loss in AACG and help to provide objective protein biomarkers to evaluate VF loss. These findings will benefit the application of the AH proteome to clinical research.