Purpose: The purpose of this study was to investigate candidate regulatory proteins associated with the pathogenesis of pterygium and to explore their potential biological roles.
Methods: Paired pterygium and conjunctival tissues from the discovery cohort (n = 10) and the validation cohort (n = 3) were subjected to proteomic analysis (label-free/4D label-free) to identify candidate differentially expressed proteins (DEPs). Exploratory clinical correlation and diagnostic analyses of DEPs were performed. Functional validation was conducted using primary conjunctival epithelial cells (PCECs) treated with C5a with or without the C5aR antagonist PMX53. Assays included evaluation of epithelial-mesenchymal transition (EMT) markers (RT-qPCR, Western blot, and immunofluorescence), migration (wound-healing assay), apoptosis (flow cytometry), and VEGFA expression.
Results: Proteomic analysis showed that C5 was consistently upregulated, and upregulated DEPs were significantly enriched in the complement and coagulation cascades. Furthermore, C5 showed a significant positive correlation with vessel density (r = 0.68, P = 0.029) and achieved an area under the curve (AUC) of 0.8 in the discovery cohort. Functional validation demonstrated that C5a promoted EMT-like marker changes in PCECs, including decreased E-cadherin and increased vimentin and fibronectin (FN). C5a treatment was also associated with enhanced cellular migration and reduced apoptosis, and these cellular changes were attenuated by the C5aR antagonist PMX53. Notably, C5a also increased VEGFA expression.
Conclusions: These findings suggest that C5a/C5aR signaling may contribute to pterygium pathogenesis by promoting EMT-related changes in conjunctival epithelial cells, supporting C5aR as a potential therapeutic target.