以抑制小胶质细胞活化为靶标探索龙血竭抗阿尔茨海默病的物质基础及作用机制

Microglia activation-induced neuroinflammation is beliveved to play a critical role in the pathology of Alzheimer's disease (AD). Therefore, it is of important significance to find safe and effective inhibitors of microglia activation. In our previous study, we have found for the first time that Chinese Dragon's Blood can sigificantly inhibit microglia activation. Further study has confirmed its effect against AD tested by using AD animal model.It has also been revealed that its inhibitory effect of microglia activation may be associated with the inhibition of ROS/MAPKs/NF-κB pathway. Based on our previous work, further investigation will be done in this project including: 1) isolation and identification of chemical compounds from Chinese Dragon's Blood by using LC-MS analysis method. 2) evaluation of the inhibitory activity of active compounds on microglia activation and further confirmation of the in vivo activity of the active compounds by using the AD animal model. 3) elucidation of mechanism and targets of the active compounds involved in the inhibitory activity on microglia activation by using both microglial cell line and APP/PS1 transgenic mice, clarification whether ROS/MAPKs/NF-κB pathway is involved in the activity of Chinese Dragon's Blood in its inhibitory effect on microglia activation and neuronal protective effect. 4) clarification of the material basis and mechanism of Chinese Dragon's Blood against AD via inhibiton of activated microglia. The purpose of this project is to provide scientific supports for the discovery of novel pharmacological activity of the traditional Chinese medicine Dragon's Blood and related material basis, as well as to establish the experimental foundation of the discovery of new drug for the treatment of AD.

小胶质细胞活化诱发的神经炎症是阿尔茨海默病（Alzheimer's disease，AD）产生和发展的重要原因，已成为抗AD药物研究的新靶标。我们新近研究首次发现名贵中药龙血竭能够显著抑制小胶质细胞活化。进一步活性导向的分离和动物实验确证了其活性成分之一LXJ05的抗AD作用及体内小胶质细胞活化的抑制作用。由此，本项目拟继续阐明龙血竭活性部位的物质基础；使用细胞水平的筛选模型评价其小胶质细胞活化的抑制活性并利用整体动物模型进行在体活性确证；综合运用分子生物学方法，以ROS/MAPKs/NF-κB通路为切入点，利用小胶质细胞系及APP/PS1转基因小鼠，阐明其活性成分如何通过对此信号通路的调控抑制小胶质细胞活化释放炎性介质、从而发挥对神经元的保护作用机制，明确其体内外分子信号调控机制的相关性，以期为揭示传统中药龙血竭抗AD这一全新的药理活性及物质基础奠定实验基础，为中药现代化提供实验依据。

小胶质细胞活化诱发的神经炎症是阿尔茨海默病（Alzheimer's disease，AD）产生和发展的重要原因，已成为抗 AD 药物研究的新靶标。我们前期研究首次发现名贵中药龙血竭能够显著抑制小胶质细胞活化。进一步活性导向的分离和动物实验确证了其活性成分之一 LXJ05 的抗 AD 作用及体内小胶质细胞活化的抑制作用。由此，本项目在前期工作基础上继续阐明龙血竭活性部位的物质基础，龙血竭二氯甲烷萃取层及乙酸乙酯萃取层为其有效部位，从上述有效部位中分离鉴定了47个单体化合物，其中3个为未见文献报道的新化合物；使用细胞水平的筛选模型评价上述单体化合物小胶质细胞活化的抑制活性，构建了龙血竭活性成分抑制小胶质细胞活化活性的构效关系，在利用整体动物模型进行在体活性确证的基础上，明确了紫檀茋为其指标性活性成分；综合运用分子生物学方法，以 ROS/MAPKs/NF-κB 通路为切入点，利用小胶质细胞系阐明了指标性活性成分紫檀茋通过对此信号通路的调控抑制小胶质细胞活化释放炎性介质、从而发挥对神经元的保护作用机制，在此基础上采用两种AD模型小鼠进一步进行体内实验，明确了其体内外分子信号调控机制的相关性。本课题为揭示传统中药龙血竭抗 AD 这一全新的药理活性及物质基础奠定实验基础，为中药现代化提供实验依据。