| Purpose:As the global myopia situation becomes increasingly severe,pathologic myopia has become one of the main causes of low vision and blindness in the world,especially in China.Complications of pathologic myopia,such as posterior scleral staphyloma,choroidal/retinal atrophy,and choroidal neovascularization in the macular region are the main causes of visual impairment.Although we are now facing such a severe situation of prevention and treatment of pathologic myopia,there is still a lack of effective clinical strategies to treat pathologic myopia at home and abroad.The main reason for this dilemma is that our understanding of the pathogenesis of pathologic myopia is not deep enough.With the rapid development of mass spectrometry and biological database technology,we try to use label-free quantitative proteomic analysis to compare the proteomic changes in aqueous humor of age related cataract patients with or without pathologic myopia.The signal transduction pathway and key proteins involved in differentially expressed proteins of pathologic myopia were obtained by bioinformatics analysis,which provided theoretical basis for exploring the pathologic mechanism and treatment of pathologic myopia.Methods:Aqueous humor samples were collected from a total of 80 age related cataract patients with or without pathologic myopia(forty in each group).Twenty samples from each group were analyzed with label-free quantitative proteomic analysis using liquid chromatography tandem mass spectrometry.The differentially expressed proteins were identified and futher validated by enzyme linked immunosorbent assay(ELISA)for the 5 randomly selected differentially expressed proteins.Function enrichment analyses were performed by Panther datadase,gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG).The protein-protein interaction network was constructed and the module analysis was performed using STRING and Cytoscape to find the hub protein.Hub protein was validated with ELISA using the second group of independent cohort consisting of twenty samples from each group and its receiver operating characteristic(ROC)curve analysis was conducted.Results:A total of 583 proteins were identified and 101 proteins were found to be differentially expressed,including 63 up-regulated proteins and 38 down-regulated proteins,which were further validated using ELISA for the 5 randomly selected differentially expressed proteins in the other twenty samples of each group.The main classifications of the differentially expressed proteins were protein-binding activity modulator(25.70%),defense/immunity protein(24.80%),protein modifying enzyme(11.40%),metabolite interconversion enzyme(8.70%),extracellular matrix protein(7.80%),transfer/carrier protein(3.80%)and so on.The bioinformatics analysis suggested that pathologic myopia is closely associated with immunity and inflammation interactions,and remodeling of extracellular matrix.Apolipoprotein A1(Apo A1)was enriched as the hub protein of the network with the highest score(MCODE score=19.085),degree(degree=23)and centrality(centrality=1.0).ROC analysis showed that Apo A1 could distinguish pathologic myopia from controls with an area under the curve(AUC] of 0.963(p < 0.001).Furthermore,the up-regulated expression trend of Apo A1 in pathologic myopia was further validated in an independent set of samples consisting of twenty subjects in each group.Conclusions:Our quantitative proteomic analysis revealed altered proteomics profiles in the aqueous humor from pathologic myopia.The findings could provide potential clues for further study on the molecular mechanisms of the pathologic myopia and developing new treatments for pathologic myopia,especially related to immunity and inflammation interactions.Apo A1 may be a potential key protein and therapeutic target in human pathologic myopia. |
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