黎药冰糖草抑制JNK通路抗胰岛β细胞凋亡特有二萜的发现及其作用机制研究

从民族药寻找作用机制特殊的有效物质是抗糖尿病新药研发的策略之一。冰糖草为海南常用黎药，资源丰富。课题组前期发现，冰糖草抗胰岛β细胞凋亡（PIβCA）有效部位富含特有的新颖二萜，且其活性与抑制JNK通路密切相关。本项目拟在前期基础上，系统获得有效部位中的二萜；评价所得二萜抗PIβCA的强弱，观察它们与JNK的相互作用，进行衍生化改造，剖析构效关系，遴选结构新、活性佳的二萜（或改造产物）；评估遴选二萜改善db/db小鼠胰岛功能的作用，观察其对胰岛细胞形态和凋亡的影响，从细胞、形态、生理、病理多层次立体确证其药效；检测遴选二萜对JNK活性的影响及对下游靶蛋白基因的表达、转录调控，结合特异性阻断剂和RNA干扰，阐释其通过抑制JNK通路发挥功效的机制。本项目的实施，将揭示冰糖草抗PIβCA的药效物质基础和作用机制，为黎药来源抗PIβCA药物研究提供类药性佳的先导化合物，为抗糖尿病民族药研究提供示范。

Finding effective agents from ethnic drugs with special mechanisms is an effective way in developing drugs for the treatment of diabetes. Scoparia dulcis was an commonly used Li National Medicine, and is abundant in natural resources. Our research group previously found that the effective fraction from Li National Medicine of Scoparia dulcis displayed significant inhibiting abilities on pancreatic islet β cells apoptosis (PIβCA). This effective fraction was rich in unique diterpenoids with anti-PIβCA effects which was closely correlated with inhibiting JNK signal pathway. Based on these previous results, this project aims to complete the following researches. To systematically obtain the diterpenoids from active fraction, evaluate the anti-PIβCA ability of obtained diterpenoids, test the binding capabilities between diterpenoids and JNK, modify the active diterpenoids, analyze the structure-activity relationship, and select diterpenoids, isolated diterpenoids from Scoparia dulcis or structurally modified diterpenoids, with unique structures and significant activities. To detect the improvement on islet function of the selected diterpenoids using diabetic db/db mice, observe its effects on PIβCA and cell morphology changes. By these investigations, the anti-PIβCA effects of the selected diterpenoids will be systematically studied on multiple angles including cell morphology, physiology, pathology, etc. To test the ability of selected diterpenoids on activating JNK and regulating the expression and transcription of downstream target proteins and genes. By combing with the specific blocker and RNA interference, this project will clarify the mechanism of selected diterpenoids on inhibiting JNK. By implementation of the project, the active substances and molecular mechanism will be announced. The lead compounds with potential of drugability will be found which will provide solid bases for development of anti-PIβCA drugs from Scoparia dulcis. This project will set an example for anti-diabetic national medicine research.