基于抑制AGEs蓄积偶联炎症损伤机制的知母黄柏药对抗糖尿病骨质疏松药效物质基础研究

糖尿病骨质疏松(DOP)是发病率最高的糖尿病并发症，晚期糖基化终末产物(AGEs)蓄积偶联炎症损伤是DOP致骨丢失的核心机制，但缺乏针对性干预手段。滋阴清热经典药对知母黄柏是抗DOP方药的核心组成且高频使用，申请人前期发现，知母黄柏药对可降低DOP大鼠AGEs蓄积、抑制炎症损伤，然而其抗DOP药效物质基础不明。据此，本课题将围绕抗AGEs蓄积偶联炎症损伤机制，先用代谢组学技术构建药对药效物质轮廓；再用非酶糖基化反应、细胞亲和色谱及网络药理学技术，针对核心机制的三个关键病理环节(AGEs蓄积、AGEs受体结合活性和AGEs炎症损伤)筛选靶成分；最后构建定量组效关系，在DOP动物及AGEs损伤成骨/破骨细胞模型上明确靶成分协同配伍关系，阐明药对抗DOP主效成分群及机制。期望从抑制AGEs蓄积偶联炎症损伤的角度阐明知母黄柏药对配伍机理，为发展化学组成与作用机制清晰的中药抗DOP新策略提供依据。

Diabetic osteoporosis (DOP) is the most common diabetic complication. Advanced glycation end products (AGEs) accumulation and the related inflammatory injury are the core reason for DOP-induced bone loss. However, there is no effective treatment for DOP based on this core mechanism of the disease. As the classic herb-pair, Zhimu-Huangbai herb-pair was always used as the important components in anti-DOP prescription. Our previous studies found that the Zhimu-Huangbai herb-pair could reduce AGEs levels and inhibit inflammatory damage in DOP rats. However, anti-DOP substances from Zhimu-Huangbai herb-pair is unclear. This project will clarify the anti-DOP substances of Zhimu-Huangbai herb-pair based on the mechanism of AGEs accumulation and inflammatory injury. Firstly, the profile of anti-DOP substances will be found by using metabolomics technology. Then, non-enzymatic glycosylation reaction, cell affinity chromatography and network pharmacology technology will be used for screening the target substances, which can regulate three key pathological processes (AGEs accumulation, AGEs receptor binding activity, and AGEs inflammatory damage) of the core mechanism. Finally, composition-activity relationship will be constructed to identify principal effective substances and the synergistic effect in DOP animals and AGEs injured osteoblast/osteoclast models. It is expected to clarify the synergistic mechanism of Zhimu-Huangbai herb-pair from the perspective of inhibiting AGEs accumulation and inflammatory injury. This project will provide a basis for developing a new strategy for DOP treatment by traditional Chinese medicine with clear chemical compositions and mechanism.