基于FOXO活性以及糖尿病果蝇模型的金芪降糖片活性组分分析

Diabetes is a common group of metabolic diseases. he inactivation of insulin function is the main cause for the high circulating glucose phenotype. The activity of Forkhead box O transcription factor (FOXO) is regulated by insulin signaling, FOXO is an important player controlling energy homeostasis via its downstream targets, many of which are sugar and lipids metabolic enzymes. The idea of this project is based on holistic strategy of Chinese traditional medicine (TCM).The innovation of this project is to integrate biochemistry, organic chemistry and genetic methods and technology. By establishing new screen strategies from natural products based on FOXO activity and fruit fly (drosophila) diabetic models, using chemical markers' fishing and knockout technology to analyze bioactive components from Jinqi Jiangtang tablets, we can get those components which can restore insulin signaling or FOXO activity, thus relieving metabolic phenotypes. Furthermore, we can analyze the mechanism how these components affect insulin signaling. Based on these study, we try to elucidate the bioactive components from Jinqi Jiangtang Tablets compound library and disclose their contribution. We can setup effective screen strategy and methods to fish out bioactive components from Chinese Herbal Compound, investigate their biological mechanisms and develop new anti-diabetes drugs.

糖尿病为常见代谢疾病，胰岛素功能的失活是导致高血糖症状的主要因素。叉头框转录因子O（FOXO）受胰岛素通路调控，通过操控糖、脂代谢相关基因的表达调控机体代谢平衡。本项目传承和创新中药整体观的主要思路，通过集成生物化学、有机化学、遗传学等多学科的方法和技术，建立基于FOXO活性和果蝇糖尿病模型的天然药物活性成分的筛选方法，利用成分群缺失/捕获技术对金芪降糖颗粒活性组分分析，筛选对胰岛素信号转导通路有影响的中药活性成分，揭示活性物质与胰岛素信号转导通路的相互作用机制。通过以上研究，旨在整体化学成分群基础上，了解金芪降糖片活性成分群及其活性贡献度，建立行之有效的中药复方生物活性物质筛选方法，探讨复方作用机制以及开发抗糖尿病新药。

建立快速、高灵敏、高选择和高分辨的现代分析技术（ UPLC-Q-TOF/MS, UPLC-IT-TOF/MS, GC-MS 等），对金芪降糖片的固有化学成分进行定性定量表征研究;建立果蝇模式生物的降糖降脂活性物质的筛选方法，发现与捕获金芪降糖片有效成分（群），明确生物碱类成分为其主要降糖降脂物质基础，使用果蝇糖尿病模型，对包括金芪降糖片在内的多味中药复方及药对进行活性筛选，发现一系列具有降糖降脂的组合。其中，以丹参、三七药对效果为佳;在细胞模型上研究FOXO 、SREBPs等转录因子在代谢平衡中的调控作用，以及该转录因子在各靶向基因的不同的条件下，是如何通过协调各种生物活性蛋白以及信号通路来调节机体代谢平衡的，并对于金芪降糖片成分及其它天然产物活性成分进行筛选，获得一批具有显著活性的先导化合物，并为相应的胰岛素通路异常导致的代谢性疾病的治疗提供新的先导化合物，为今后化学修饰合成高效、低毒的Ⅱ型糖尿病药物奠定基础。期间，已发表SCI论文5篇，其中一篇影响因子>5，申请专利4项，授权一项；培养硕士生3名，博士生1名。