基于细胞膜键合磁性碳球垂钓技术和网络药理学探究三子丸抗骨质疏松药效物质及作用机制

细胞膜垂钓技术是近年来用于中药（复方）活性成分快速筛选的前沿热点技术，但其主要不足在于细胞膜容易从磁性固载基质上脱落，导致垂钓复合材料的稳定性较差。三子丸是《千金要方》所载的经典补肾中药方剂，由蛇床子、菟丝子、五味子三味药组成，申请人前期确证三子丸能显著减缓去卵巢骨质疏松（OP）大鼠的骨丢失，且对成骨细胞增殖的促进和对破骨细胞分化的抑制作用显著高于单味药及两味药组合，但其抗OP的药效物质基础尚不明确。有鉴于此，本项目在前期研究的基础上，拟分别建立大鼠成骨和破骨细胞膜键合磁性碳球垂钓技术，显著提升传统细胞膜垂钓技术的稳定性和重复性，结合原方成分分析及药效学实验，高效发掘出三字丸抗OP的药效物质；再通过网络药理学研究和生物学验证，确认三字丸药效物质的作用靶点和信号通路，为阐明三子丸多成分、多靶点整合防治OP的作用机制奠定良好的基础，也为其他中药复方药效物质基础及作用机制的研究提供新颖的思路。

Recently, cell membrane (CM)-fishing technique has become the focus and gain frontier position in the rapid screening of active constituents from traditional Chinese medicine (compound). Its main deficiency is the poor lifespan of fishing materials, due to the fall-off of CM from magnetic substrates. Sanziwan is a classic Chinese kidney-tonifying recipe recorded in "Thousand-Golden-Prescriptions", which consists of fructus cnidii, Cuscuta chinensis, and Schisandra chinensis. Our previous work has confirmed that Sanziwan can dramatically prevent the bone loss of ovariectomized rats with osteoporosis (OP), and has more effect on the promotion of osteoblast proliferation and the inhibiting of osteoclast differentiation than that of its ingredients. However, the pharmacodynamic substance foundation of Sanziwan on OP remains unclear. In light of above facts, the osteoblast and osteoclast membrane-bonded magnetic carbon sphere fishing techniques will be developed based on our previous work, thus significantly enhancing the stability and repeatability of traditional CM-fishing methods. Then, the anti-osteoporosis active components of Sanziwan will be efficiently discovered through the combination of developed techniques, the Sanziwan component analysis and pharmacodynamic experiment. After that, the effect targets and signaling pathways of these components will be further investigated by the network pharmacology research and biological evaluation. These researches will lay a good foundation for illustrating the multi-component and multi-target action mechanism of Sanziwan in the integrate control of OP, and also provide a novel methodology for studying the pharmacodynamic substance foundation and action mechanism of other Chinese medicine compound.