Abstract：
Oridonin, a natural terpenoid isolated from the leaves of Isodon rubescens (Hemsley) H.Hara, is widely used in oriental medicine for its anticancer properties across various cancer types. Despite its prevalent use, the toxic effects of oridonin on male reproduction, particularly its impact on sperm functions and the mechanisms involved, are not well understood. This study aimed to explore the effects and underlying mechanisms of oridonin on sperm functions. We initially treated Duroc boar spermatozoa with varying concentrations of oridonin (0, 5, 50, 75, 100, and 150 µM) and incubated them to induce capacitation. We then assessed cell viability and several sperm functions, including sperm motility and motion kinematics, capacitation status, and ATP levels. We also analyzed the expression levels of proteins associated with the phosphatidylinositol 3-kinase (PI3K)/phosphoinositide-dependent kinase-1 (PDK1)/protein kinase B (AKT) signaling pathway and phosphotyrosine proteins. Our results indicate that oridonin adversely affects most sperm functions in a dose-dependent manner. We observed significant decreases in AKT, p-AKT (Thr308), phosphatase and tensin homolog (PTEN), p-PDK1, and p-PI3K levels following oridonin treatment, alongside an abnormal increase in phosphotyrosine proteins. These findings suggest that oridonin may disrupt normal levels of tyrosine-phosphorylated proteins by inhibiting the PI3K/PDK1/AKT signaling pathway, which is crucial for cell proliferation, metabolism, and apoptosis, thus potentially harming sperm functions. Consequently, we recommend considering the reproductive toxicity of oridonin when using it as a therapeutic agent.
摘要：
Oridonin,一种天然萜类化合物，从冬凌草（Hemsley）H.Hara的叶子中分离出来，由于其在各种癌症类型中的抗癌特性，被广泛用于东方医学。尽管它的普遍使用，冬凌草甲素对男性生殖的毒性作用，特别是它对精子功能和相关机制的影响，不是很了解。本研究旨在探讨冬凌草甲素对精子功能的影响及其机制。我们最初用不同浓度的冬凌草甲素（0、5、50、75、100和150µM）处理杜洛克公猪精子，并将其孵育以诱导获能。然后我们评估了细胞活力和几种精子功能，包括精子运动和运动运动学，获能状态，ATP水平。我们还分析了与磷脂酰肌醇3-激酶（PI3K）/磷酸肌醇依赖性激酶-1（PDK1）/蛋白激酶B（AKT）信号通路和磷酸酪氨酸蛋白相关的蛋白质的表达水平。我们的结果表明，冬凌草甲素以剂量依赖性方式对大多数精子功能产生不利影响。我们观察到AKT显著下降，p-AKT(Thr308),磷酸酶和张力蛋白同源物（PTEN），冬凌草甲素治疗后p-PDK1和p-PI3K水平，伴随着磷酸酪氨酸蛋白的异常增加。这些发现表明冬凌草甲素可能通过抑制PI3K/PDK1/AKT信号通路破坏酪氨酸磷酸化蛋白的正常水平，这对细胞增殖至关重要，新陈代谢，和细胞凋亡，从而可能损害精子功能。因此,我们建议在使用冬凌草甲素作为治疗剂时考虑其生殖毒性。
