Abstract

Sal alleviates H₂O₂-induced oxidative damage and ECM deposition in HTMCs. • The TNF signaling pathway is a key upstream pathway mediating the protective effects of Sal. • Sal upregulates MMP3 expression via the TNF signaling pathway, thereby alleviating oxidative damage and ECM deposition in HTMCs. • In vivo experiments confirm that Sal effectively reduces intraocular pressure and ameliorates retinal ganglion cell injury by regulating the TNF/MMP3 axis. Background The functional integrity of human trabecular meshwork cells (HTMCs) is essential for regulating intraocular pressure (IOP). In primary open-angle glaucoma (POAG), pathological stimuli induce oxidative damage and promote excessive extracellular matrix (ECM) deposition in HTMCs, thereby hindering aqueous humor outflow. Salidroside (Sal), a compound with diverse pharmacological activities, has been reported to inhibit oxidative stress and reduce ECM accumulation. This study aimed to investigate whether Sal exerts its antioxidant effects by regulating matrix metalloproteinase 3 (MMP3) expression via the TNF signaling pathway. Methods Potential targets of salidroside and glaucoma were predicted using SwissTargetPrediction and GeneCards online databases, and overlapping targets were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on these intersecting targets. Meanwhile, the glaucoma GEO dataset (GSE27276) was downloaded for differential gene expression analysis, and the results were further compared with the intersecting targets to identify key genes. An oxidative damage model of HTMCs was established using hydrogen peroxide (H₂O₂) induction. Cell viability was assessed using the MTT assay to determine the appropriate H₂O₂ concentration for modeling, as well as the safe dosage and treatment duration of Sal. Oxidative stress markers were measured as follows: superoxide dismutase (SOD) and malondialdehyde (MDA) levels were determined by enzym