基于网络药理学和活性成分群理论的藏药旺拉CE 抗AD的PCB/RM分子网络机制研究

Alzheimer's disease (AD) is a major disease with a clinical lack of satisfactory drugs and Innovative drug research is of great significance.The traditional Tibetan medicine Wangla (Coeloglossumviridevar. Bracteatum) regulating body elements, invigorating qi and blood, tranquilize and increasing intelligence，and have anti-Alzheimer's disease curative effect. The applicant's previous studies have proved that the active components group of Tibetan medicine Wangla (Coeloglossumviridevar. Bracteatum extract, CE) can resist Alzheimer's disease with obvious features and promising application prospects. However, but the key molecular mechanism of CE anti-AD is unclear, so it has not become mechanism clear national innovative drug. This project is intended to be based on previous research, using AD cell model by Aβ mediated and APP/PS1 transgenic animal model, the effect of CE on pathological changes, inflammatory response and learning and memory ability of neurons in the process of AD was discussed in vitro and in vivo；To explore the influence of CE on CAMP-PKA-CREB-BDNF and TNF-a/ RIPK1/RIPK3-MLKL signaling pathway(PCB/RM molecular network) and the key genes BDNF or RIPK; Screening CE highly active monomer HCE and its action targets, Using RNAseq analysis technique, we study the CE protection on AD on the whole transcriptome and key action sites，specially focus on the PCB/RM molecular network; to provide key data for Innovative pioneering drugs with clearly composition, target spot and mechanism. This study also provides a reference model for anti-AD innovation national drug research.,to elucidate the pharmacodynamic material basis of the national medicine wangle anti-AD, and to provide scientific and technological support for the development of ethnic medicine. anti-AD advantages and characteristics.

阿尔茨海默病(AD)属重大疾病，临床缺乏满意药物和疗法。传统藏药旺拉调三素、固三根、安神增智，抗AD疗效好。申请人以往多项课题研究证明藏药旺拉活性成分群CE可抗AD，极具应用前景。但因CE抗AD的关键分子机制不清，影响其成为机制清楚民族创新药。拟在前期研究基础上，利用Aβ介导AD细胞模型和APP转基因小鼠模型，在离体和在体水平研究CE在AD发生过程中对神经元病理性改变、突触可塑性、炎症反应和动物学习记忆能力的影响；基于网络药理学关键基因变化系统研究其对PCB/RM分子网络的影响；筛选其高效活性单体HCE及作用靶点；并结合RNAseq技术在转录组水平整体地分析CE调控AD的PCB/RM分子网络和关键作用位点，为获得成分清楚、靶点明确、机制清晰创新先导药提供关键数据。本研究阐释藏药旺拉抗白脉病AD的药效物质基础，为发挥民族药抗AD优势和特色提供科技支撑，也为抗AD创新民族药研究提供参考模式。

阿尔茨海默病(AD)属重大疾病，临床缺乏满意药物和疗法。珍贵藏药旺拉（ Coeloglossum viride (L) Hartm. var. bracteatum (Willd.) Richter）是一种在藏文化中具有多种用途的食药两用传统植物补品，具有补阳、益气、安神、益智、长寿等功效。藏蒙医药常用于治疗阳痿、遗精、贫血、失眠等，也是藏医治疗白脉病隆症的常用药材，疗效确切，历史悠久，已经在藏区人工大面积繁育。我们以此为研究的依据，利用分子、细胞、形态学手段，在多种动物模型和细胞模型上对其活性成分群（Coeloglossumviridevar. Bracteatum extract, CE）及特征性活性成分对神经系统功能的调节和神经退行性疾病的治疗作用及分子机制，尤其是在多种AD动物和细胞模型上进行了多方面的研究。研究证实了CE 在 AD 等发病过程中的神经保护功能及其基于神经营养/神经炎症分子网络的分子机制， 尤其是对PCB/RM 分子网络机制的影响。我们的研究发现证明CE及其活性单体对APP/PS双转基因5X FAD小鼠、侧脑室注射Aβ25-35致痴呆模型小鼠、AlCl3/ D -半乳糖致认知障碍小鼠和MPTP介导的急性帕金森病（PD）小鼠有较好的脑功能改善和保护作用。CE及其单体能清除5XFAD小鼠脑内的Aβ斑块改善认知功能的障碍，改善Aβ毒性介导的痴呆模型小鼠和AlCl3/ D -半乳糖致衰老小鼠的认知功能的障碍和改善MPTP介导的急性PD小鼠的运动障碍。CE可上调神经营养因子BDNF和FGF2水平，调节PI3K-AKT/MAPK-ERK信号通路，抑制TBK1负调控的RIPK1/RIPK3/MLKL坏死性凋亡信号通路，抑制Notch/ hes1信号轴和PKM2/HIF1a信号轴介导的小胶质细胞的激活而抑制神经炎症，提高脑内能量代谢水平。CE作用机制相对明朗，可能成为治疗白脉病的重要民族药物，值得进一步深入研究并开发应用。这也为传统医药多成分、多靶点、多通路、多机制对疾病的药理研究和治疗应用开辟了新的途径。. 在国家自然科学基金委的大力资助下，课题组针对民族医药开展攻关研究。课题组按申请书内容认真实施，发表10篇高水平论文，其中包括Top期刊论文2篇（中科院大类一区）、中科院大类二区论文7篇和中文核心期刊1篇。超额完成任务。