DataSheet7_Integrative proteomics and metabolomics approach to identify the key roles of icariin-mediated protective effects against cyclophosphamide-induced spermatogenesis dysfunction in mice.CSV

The alkylating antineoplastic agent cyclophosphamide (CP) is known to be toxic to the male reproductive system, but there are no effective prevention or treatment options. The flavonoid icariin (ICA), which is used in Chinese medicine, has been shown to have a number of biological functions, including testicular protection. The current study looked into the protective effects of ICA in preventing CP-induced spermatogenesis dysfunction. The current study looked into the role of ICA in preventing testicular dysfunction caused by CP. For 5 days, healthy adult mice were given saline or a single dose of CP (50 mg/kg) intraperitoneally (i.p). For the next 30 days, mice were given ICA (80 mg/kg) by gavage. Animals were euthanized 12 h after receiving ICA, and testes were removed for biochemical, histopathological, sperm evaluation, and transmission electron microscope analysis (TEM). We also investigated the potential biological effects of ICA on CP-induced spermatogenesis dysfunction in mice using an integrated proteomic and metabolomic approach. The levels of 8309 proteins and 600 metabolites were measured. The majority of the differential proteins and metabolites were found to be enriched in a variety of metabolic pathways, including the PI3K-Akt signaling pathway, necroptosis, the mTOR signaling pathway, glycerophospholipid metabolism, and ABC transporters, implying that ICA may have molecular mechanisms that contribute to CP-induced spermatogenesis dysfunction in the testis. Taken together, these findings show that ICA effectively reduces testis injury, implying that ICA may have a role in male infertility preservation.

烷化类抗肿瘤药物环磷酰胺（cyclophosphamide, CP）已知可对雄性生殖系统造成毒性损伤，但目前尚无有效的预防或治疗方案。中药活性成分淫羊藿苷（icariin, ICA）已被证实具备多种生物学功能，其中包括睾丸保护作用。本研究旨在探究ICA对环磷酰胺诱导的精子生成功能障碍的保护作用，以及其对CP所致睾丸功能损伤的干预效果。实验中，健康成年小鼠连续5天接受腹腔注射生理盐水，或单次腹腔注射50mg/kg的环磷酰胺；随后连续30天以灌胃方式给予80mg/kg的ICA。末次给药12小时后处死小鼠，摘取睾丸组织进行生化检测、组织病理学分析、精子功能评估以及透射电子显微镜（transmission electron microscope, TEM）观察。此外，本研究采用蛋白质组学与代谢组学联合分析策略，探究ICA对小鼠CP诱导的精子生成功能障碍的潜在调控机制。本次实验共检测到8309种蛋白质与600种代谢物，经富集分析发现，绝大多数差异蛋白与代谢物显著富集于多条核心代谢通路，包括PI3K-Akt信号通路、坏死性凋亡、mTOR信号通路、甘油磷脂代谢以及ABC转运蛋白通路，提示ICA或通过调控上述分子通路，改善环磷酰胺诱导的睾丸精子生成功能障碍。综上，本研究结果表明ICA可有效减轻睾丸组织损伤，提示其在雄性不育的临床防护中具备潜在应用价值。