Abstract：
Epimedii Folium (EF), a commonly used herbal medicine to treat osteoporosis, has caused serious concern due to potential hepatotoxicity. Until now, its intrinsic hepatotoxic mechanism and hepatotoxic ingredients remain unclear. Here, a novel high-throughput approach was designed to investigate the intrinsic hepatotoxic of EF. High-content screen imaging (HCS) and biochemical tests were first performed to obtain the cytotoxicity parameter matrix of 17 batch EF samples. EF-treated alpha mouse liver 12 (AML12) cells showed increased reactive oxygen species (ROS), reduced glutathione (GSH) and mitochondrial membrane potential (MMP), and apoptosis and cholestasis were further observed. Network toxicology predicted that EF-triggered hepatotoxiciy was involved in transcription factor (TF) activity. The FXR expression, screened by a TF PCR array, exhibited down-regulation following EF extract administration. Moreover, EF inhibited bile acid (BA) metabolism pathway in an FXR-dependent manner. Pearson correlation between the cytotoxicity parameter matrix and quantification feature table obtained from UHPLC-QTOF data of EF suggested 7 prenylated flavonoids possessed potent hepatotoxicities and their cytotoxicity order was further summarized. The transcriptional repression effects of them on FXR were also verified. Collectively, our findings indicate that FXR is probably responsible for EF-induced hepatotoxicity and prenylated flavonoids may be a major class of hepatotoxic constituents in EF.
摘要：
淫羊藿(EF),一种常用的治疗骨质疏松症的草药，由于潜在的肝毒性引起了严重的关注。直到现在,其内在的肝毒性机制和肝毒性成分尚不清楚。这里,我们设计了一种新的高通量方法来研究EF的内在肝毒性.首先进行了高含量屏幕成像（HCS）和生化测试，以获得17批EF样品的细胞毒性参数矩阵。EF处理的α小鼠肝脏12（AML12）细胞显示出增加的活性氧（ROS），还原型谷胱甘肽（GSH）和线粒体膜电位（MMP），进一步观察到细胞凋亡和胆汁淤积。网络毒理学预测EF触发的肝毒性与转录因子（TF）活性有关。FXR表达式，通过TFPCR阵列筛选，在EF提取物施用后表现出下调。此外，EF以FXR依赖性方式抑制胆汁酸（BA）代谢途径。从EF的UHPLC-QTOF数据获得的细胞毒性参数矩阵与定量特征表之间的Pearson相关性表明，7个异戊烯黄酮具有有效的肝毒性，并进一步总结了它们的细胞毒性顺序。还验证了它们对FXR的转录抑制作用。总的来说，我们的发现表明FXR可能是EF诱导的肝毒性的原因，异戊烯化类黄酮可能是EF中的一类主要肝毒性成分。
