基于17β-HSD2/CYP450s-GSTs的补骨脂“效毒一体”物质基础及作用机制研究

阐明中药效/毒成分的分子机制是保障安全有效用药亟待解决的核心环节。补骨脂是治疗骨质疏松的常用有毒中药，但目前其效毒研究尚缺乏基于直接作用靶标的证据链。我们结合前期研究提出“补骨脂中香豆素类成分可能代谢形成环氧中间产物，其既可能与17β-HSD2直接加成，抑制雌激素向雌酮转化，发挥补肾壮骨的效应；也可能通过共价结合CYP450s、GSTs影响药物代谢导致肝毒性，产生效毒一体现象”假说。围绕此假说，本项目拟通过效/毒作用导向的UPLC-Q/TOF筛选系统辨析补骨脂关键效毒成分；并通过效毒成分分子探针进行靶点捕获；进而整合表面等离子共振、激光共聚焦共定位及定向蛋白组学等方法从“分子-细胞-整体动物”多层次阐明效毒分子与靶蛋白的相互作用关系，构建“关键成分-直接靶标-效/毒作用”的证据链，初步揭示补骨脂效毒一体现象科学内涵，为补骨脂安全有效用药提供理论依据，并为“有毒”中药研究提供新的思路。

To clarify the molecular mechanism of the efficacy / toxic components of traditional Chinese medicine is the core point that needs to be urgently solved to ensure effective and safe medication in clinic. Psoraleae Fructus is a toxic traditional Chinese medicine for the treatment of osteoporosis, but its efficacy and toxicity research lacks a chain of evidence based on direct-acting targets. Based on previous research, we proposed that "coumarin components in Psoraleae Fructus may be metabolized to form epoxy intermediates, which may directly add to 17β-HSD2, inhibit the conversion of estrogen to estrone, and exert the effect of nourishing the kidney and strengthening the bone; it is also possible to influence drug metabolism through covalently combining CYP450s and GSTs to lead to hepatotoxicity. These mechanisms might cause a phenomenon of effect-toxicity integration of Psoraleae Fructus". Based on this hypothesis, the project intends to identify the key efficacy / toxic components of Psoraleae Fructus through the UPLC-Q / TOF screening system and capture the target through the molecular probes of the key components; and then integrate the methods of surface plasmon resonance, laser confocal co-localization and directed proteomics to elucidate the interaction between the key components and target proteins at multiple levels of "molecule-cell-animal" to construct an evidence chain of "key components-direct target-efficiency / toxicity" of Psoraleae Fructus. To initially reveal the scientific connotation of the phenomenon of effect-toxicity integration, provide a theoretical basis for the safe and effective use of Psoraleae Fructus, and provide new ideas for the research of "toxic" Chinese medicine.