Integrating Epigenetics, Proteomics, and Metabolomics to Reveal the Involvement of Wnt/β-catenin Signaling Pathway in Oridonin-Induced Reproductive Toxicity

Oridonin is the primary active ingredient in traditional Chinese medicine Rabdosia rubescens, displaying various pharmacological activities such as anti-inflammatory, anti-tumor, and antibacterial effects. It is widely used in the clinical treatment of acute and chronic pharyngitis, tonsillitis, and bronchitis. Nevertheless, the reproductive toxicity of oridonin significantly restricts its clinical application, with the exact mechanism remaining unclear. This study aimed to investigate the mechanism of oridonin-induced damage to HTR-8/SVneo cells. By integrating epigenetics, proteomics, and metabolomics approaches, the mechanisms of oridonin-induced reproductive toxicity were discovered and confirmed through fluorescence imaging, RT-qPCR, and Western blotting. Experimental findings indicated that oridonin altered m6A levels, gene and protein expression levels, along with metabolite levels within the cells. Additionally, oridonin triggered oxidative stress and mitochondrial damage, leading to a notable decrease in WNT6, β-catenin, CLAUDIN-1, and ZO-1 protein levels. This implies that the inhibition of the Wnt/β-catenin pathway and disruption of tight junctions may be attributed to the cytotoxicity induced by oridonin and mitochondrial dysfunction, ultimately resulting in damage to HTR-8/SVneo cells.