Cassia polysaccharide regulates the effect of PAX6 low expression on myopia through the Wnt/β-catenin pathway

Abstract Objective: Aimed to explore the protective effects of cassia polysaccharides on myopia by examining their influence on ARPE-19 cells with reduced PAX6 expression. Methods: ARPE-19 cell line with diminished PAX6 expression was established using a lentiviral approach and addition of XAV-939, an inhibitor of the Wnt/β-catenin. We assessed the expression of genes and proteins involved in the Wnt/β-catenin, scleral remodeling, and cell cycle regulation following treatment with cassia polysaccharides. Gene and protein expression were quantified using RT-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 an MOI 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 the PAX6-shRNA and XAV-939 but remained unchanged in the cassia polysaccharide. Scleral remodeling markers, including MMP-2 and TGF-β, were elevated, and COL1A1 was decreased in the PAX6-shRNA, with no significant changes observed in the cassia polysaccharide. Cell cycle analysis indicated reduced CDK1 and PCNA levels in the PAX6-shRNA, with cassia polysaccharides showing no significant effect. Scratch assay results demonstrated slower wound healing in the PAX6-shRNA compared to controls over 72h, with no significant differences observed in the cassia polysaccharide. Conclusion: Cassia polysaccharides may mitigate ARPE-19 cell damage induced by low PAX6 expression through modulation of the Wnt/β-catenin, potentially slowing the progression of myopia and offering a protective effect on vision.