| Objective Age-related macular degeneration is the leading cause of blindness among older people in developed countries.As a type of late stage of the disease,wet age-related macular degeneration accounts for only 10% of the cases,but it is responsible for 90% of legal blindness due to AMD.Since current studies have not yet fully identified the pathogenic factors and related mechanisms of wet age-related macular degeneration,we hope to compare the difference between patients and normal subjects by whole proteomics detection of aqueous humor proteins and to explore the influence of intravitreal anti-vascular endothelial growth factor therapy on the composition of aqueous humor proteins in patients.Methods We performed full proteomic detection of aqueous humor samples obtained from patients with wet age-related macular degeneration before and after Conbercept treatment and cataract patients as a control group before the surgery.Then the three groups of identified aqueous humor proteins were analyzed and compared by GO functional annotation and KEGG signaling pathway.Finally,the differentially expressed proteins in aqueous humor before and after treatment were verified by Western blot in the laser-induced choroidal neovascularization mouse model.Results We identified a total of 637 proteins from all aqueous humor samples by whole proteomics.Overall,although there is no significant difference in the results of GO functional analysis in the three groups of proteins,we can still find differences in the signaling pathways in which these proteins are involved.Compared with the control group,there were more proteins located in hypoxia inducible factor-1 related signaling pathway and fewer proteins located in glycolysis signaling pathway in aqueous humor proteins of patients with wet age-related macular degeneration before treatment.Compared with patients’ aqueous humor proteins without intravitreal injection of Conbercept,the number of proteins located in the cell adhesion molecule signaling pathway in the aqueous humor of the patients after treatment was significantly higher.Then we screened the proteins,angiopoietin-related protein 7 and Dickkopf-related protein 3,which expressed statistically different in aqueous humor before and after treatment,and further verified them by animal experiments.It was found that the changes of protein expression in mouse model were consistent with the results of proteomics in human specimens.Conclusions Our study described the characteristics of aqueous humor proteins profile in patients with wet age-related macular degeneration using wholeproteomics techniques,confirming the effects of disease and drug therapy on aqueous humor proteins.We found that the pathological changes caused by hypoxia may play an important role in the disease-related pathogenesis,and the effect of treatment on aqueous humor protein profile is mainly reflected in the changes of some proteins and signaling pathways. |
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