线粒体是中药拮抗阿霉素心脏毒性的重要靶点，是活性组分筛选的良好载体。针对四逆汤拮抗阿霉素心脏毒性药效物质不清，机制不明的问题，在证实四逆汤存在直接作用于心肌细胞线粒体的组分和前期工作基础上，采用线粒体膜为活性组分和靶标识别载体，建立基于线粒体识别-二维色谱技术的中药活性组分筛选新方法，鉴定四逆汤线粒体膜结合组分，评价其拮抗阿霉素心脏毒性的活性；采用药物亲和反应靶标稳定技术，鉴定并解析四逆汤在线粒体膜上的结合靶蛋白；通过分子对接模拟，表征线粒体膜结合组分与结合靶蛋白的对应关系。遴选制备3-5种结合组分，采用siRNA干扰等实验手段，在细胞分子水平进行靶点反向试验验证，明确四逆汤拮抗阿霉素心脏毒性组分与靶标的对应关系。通过本研究，构建基于线粒体识别的中药“多组分-多靶标”关联分析的新方法，有望为拮抗阿霉素心脏毒性的新药开发提供活性先导化合物和新靶点，这也对治疗线粒体疾病的新药研究具有重要意义。

Mitochondria is an important target of Traditional Chinese medicine (TCM) against doxorubicin-induced cardiotoxicity, and it is a good recognizer for the screening of the active components from TCM. To focus on the problems that the effective components and mechanisms of Sini decoction (SND) against doxorubicin-induced cardiotoxicity are unclear, we take the mitochondrial membrane as recognizer to screen the bioactive components and targets of TCM, based on the preliminary result that some components in SND can directly target mitochondria of cardiomyocytes. In this study, we will establish a new method for the screening of the bioactive components in TCM based on the combination of the mitochondria recognition with comprehensive two-dimensional liquid chromatography, and then identify the binding components of SND on mitochondrial membrane and evaluate their bioactivities against doxorubicin-induced cardiotoxicity. Meanwhile, by using the drug affinity responsive target stability (DARTS) technique, we will identify their target protein on mitochondrial membrane. Then, we further perform molecular docking and molecular dynamics simulation to profile the corresponding relationship between the binding components and the binding targeted proteins on mitochondrial membrane. Finally, we choose three to five binding components and perform the validation of the targeted proteins on the cell model and the molecular level using siRNA and surface plasmon resonance (SPR) technologies. The relationship between the components and the targets against doxorubicin-induced cardiotoxicity will further be confirmed. By this study, we will establish a new approach to study the multicomponent-multitarget relationship based on mitochondria recognition. The results are expected to provide the active lead compounds and new targets for the new drug development against doxorubicin-induced cardiotoxicity. It is also an important significance on the discovery of new drug to treat mitochondrial diseases .