Cassia polysaccharides can regulate the effect of low PAX6 expression on the function of ARPE-19 cells through the Wnt/β-catenin pathway.

Purpose: To explore the protective effects of cassia polysaccharides on myopia by examining their influence on ARPE-19 cells with reduced PAX6 expression. Methods: The ARPE-19 cell line with diminished PAX6 expression was established using a lentiviral approach and the addition of XAV-939, an inhibitor of Wnt/β-catenin. We assessed the expression of genes and proteins involved in Wnt/β-catenin, scleral remodeling, and cell cycle regulation following treatment with cassia polysaccharides. Gene and protein expression were quantified using reverse transcription PCR and western blot analyses, respectively. Additionally, the migratory capabilities of these cells were evaluated using a scratch assay. Results: Optimal transduction was achieved with a multiplicity of infection of 20, successfully generating a stable ARPE-19 cell line with low PAX6 expression. Cassia polysaccharides did not significantly alter the expression of Wnt2 compared to control groups. Similarly, when treated with XAV-939, β-catenin levels were modified in PAX6-shRNA and XAV-939 but remained unchanged in the cassia polysaccharides. Scleral remodeling markers, including MMP-2 and TGF-β, were elevated, and COL1A1 was decreased in PAX6-shRNA, with no significant changes observed in the cassia polysaccharides. Cell cycle analysis indicated reduced cyclin-dependent kinase 1 and proliferating cell nuclear antigen levels in PAX6-shRNA, with cassia polysaccharides showing no significant effect. Scratch assay results demonstrated slower wound healing in PAX6-shRNA compared to controls over 72 h, with no significant differences observed in the cassia polysaccharides. Conclusions: Cassia polysaccharides may mitigate ARPE-19 cell damage induced by low PAX6 expression through modulation of Wnt/β-catenin, potentially slowing the progression of myopia and offering a protective effect on vision.