Author(s): Luo Jincheng*, Li Doudou, Li Peixuan, Li Jialin, Sun Xuewei, Wang Xue, Qiu Hongbin
Objective: Aimed to explore the protective effects of cassia polysaccharides on myopia by examining their influence on Adult Retinal Pigment Epithelium-19 (ARPE-19) cells with reduced Paired Box Protein (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 family member 2 (Wnt/β-catenin, scleral remodeling and cell cycle regulation following treatment with cassia polysaccharides. Gene and protein expression were quantified using Reverse Transcription Polymerase Chain Reaction (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 a Multiplicity of Infection (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 Matrix Metalloproteinase-2 (MMP-2) and Transforming Growth Factor-β (TGF-β), were elevated and Collagen Type 1 Alpha 1 (COL1A1) was decreased in the PAX6-shRNA, with no significant changes observed in the cassia polysaccharide. Cell cycle analysis indicated reduced Cyclin-Dependent Kinase 1 (CDK1) and Proliferating Cell Nuclear Antigen (PCNA) levels in the PAX6-shRNA: Paired box protein 6-short hairpin ribonucleic acid (PAX6- shRNA), with cassia polysaccharides showing no significant effect. Scratch assay results demonstrated slower wound healing in the PAX6-shRNA compared to controls over 72 h 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.
