冬凌草甲素是中药冬凌草的主要活性成分，具有抗炎、抗肿瘤、抗菌等作用。临床广泛用于急、慢性咽炎、扁桃体炎、支气管炎的治疗。然而，冬凌草甲素因其生殖毒性而使其临床应用受到极大限制，且其具体机制尚不清楚。本研究的目的是探讨冬凌草甲素诱导 HTR-8/SVneo 细胞损伤的机制。通过整合表观遗传学、蛋白质组学和代谢组学方法，通过荧光成像、RT-qPCR 和蛋白质印迹发现并证实了冬凌草甲素诱导的生殖毒性机制。实验结果表明冬凌草甲素改变了 m6A 水平、基因和蛋白质表达水平以及细胞内的代谢水平。此外，冬凌草甲素还会引发氧化应激和线粒体损伤，导致 WNT6、β-连环蛋白、CLDN1、CCND1 和 ZO-1 蛋白水平显着下降。这表明冬凌草甲素诱导的细胞毒性和线粒体功能障碍可能导致Wnt/β-catenin信号通路的抑制和紧密连接的破坏，最终导致HTR-8/SVneo细胞的损伤。

Oridonin is the primary active component in the traditional Chinese medicine Rabdosia rubescens, displaying anti-inflammatory, anti-tumor, and antibacterial effects. It is widely employed in clinical therapy for acute and chronic pharyngitis, tonsillitis, as well as bronchitis. Nevertheless, the clinical application of oridonin is significantly restricted due to its reproductive toxicity, with the exact mechanism remaining unclear. The aim of this study was to investigate the mechanism of oridonin-induced damage to HTR-8/SVneo cells. Through the integration of epigenetics, proteomics, and metabolomics methodologies, 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, CLDN1, CCND1, and ZO-1 protein levels. This implied that the inhibition of the Wnt/β-catenin signaling pathway and disruption of tight junction might be attributed to the cytotoxicity induced by oridonin and mitochondrial dysfunction, ultimately resulting in damage to HTR-8/SVneo cells.