基于FGF21/FGFR1/KLB信号通路的金芪降糖片降糖活性物质分离及其作用机制

JinqiJiangtang tablets compose of numerous compounds. It is unknown which componds responsible for the hypoglycemic effect and which signal pathway these compounds involved in. These questions become bottleneck for further research and application of the medicine. My previous work found that JinqiJiangtang tablets could activate FGF21/FGFR1/KLB pathway, and we identified four compounds which could interact with fibroblast growth factor receptor 1 (FGFR1) by Biomolecular Interaction Analysis (Biacore). This finding points out direction for identification of the functional molecule and its mechanism of action. In this study, applicant plans to isolate the responsible molecule(s) by Biacore, digital simulation docking, protein mutation and other techniques, using FGFR1 and its co-receptor βklotho (KLB) as bait. The glucose regulation mechanism of the isolated molecule will be investigated using in vitro and in vivo models to provide new idea and evidence for further development and application of the JinqiJiangtang tablets.

中成药金芪降糖片化学成分众多,但具体哪种或哪些成分通过哪种途径发挥降糖效应尚未完全清楚，成为阻碍该药物进一步研发和应用的瓶颈问题。前期研究发现金芪降糖片具有激活FGF21/FGFR1/KLB通路的能力，利用生物分子互作分析平台检测（Biacore），发现金芪降糖片中存在4种可亲和成纤维细胞生长因子受体1（FGFR1）的分子，为进一步阐明金芪降糖片的药效物质基础和分子机制指明了全新的方向。申请人以此为切入点，利用Biacore、数字模拟对接、蛋白突变等技术，以FGFR1蛋白及辅助受体βklotho（KLB）为钓饵，从金芪降糖片中钓取所有可与钓饵结合的活性物质并进行分离鉴定，再通过体外和体内模型深入研究其调控血糖的效应机制，为金芪降糖片进一步研发、利用提供全新思路和实验依据。

金芪降糖片由黄连、黄芪和金银花三味传统中草药组成。做为治疗糖尿病的中药处方药，金芪降糖片广泛用于糖尿病及其它代谢疾病的临床治疗，但其降糖机制尚不明确。本研究利用分子互作技术，以成纤维细胞生长因子受体1为靶标，从金芪降糖片中垂钓出可与成纤维细胞生长因子受体1(FGFR1)蛋白特异性亲和的活性分子巴马汀。经细胞和动物实验验证，与巴马汀可刺激FGFR1及其底物成纤维生长因子受体底物2（FRS2）磷酸化，并促进葡萄糖转运蛋白1（GLUT1）表达，起到促进HepG2细胞糖吸收和降低糖尿病小鼠血糖的效果， 以上结果证实金芪降糖片降糖功效部分依赖于巴马汀介导的FGFR1途径。本研究进一步阐明了金芪降糖片药效物质基础和作用机制，揭示了巴马汀降糖的作用靶点和新机制，为巴马汀药理作用和机制进一步研究提供了新的思路。