miR-148a-3p特异性调控RAGE/α7nAChR通路及逆转AD神经炎症损伤的机制研究

The pathophysiology of AD is very complicated, and the no-coding genes and other epigenetic and transcription regulated element may play an important role. Based on the previous research, we found that miR-148a-3p is down-regulated in AD mouse brain, which is negative connected with neuroinflammatory reaction. We suggested that the target gene of miR-148a-3p is RAGE, and that miR-148a-3p can be specifically regulated by inflammatory signal pathway proteins, such as RAGE and α7nAChR. Recently, we found that the extracts from Dracocephalum Moldavica L could prevent AD and the active compounds from Dracocephalum Moldavica L have an important influence on miR-148a-3p/RAGE signal pathway. Up to now, the mechanism of miR-148a-3p regulating RAGE/α7nAChR signal pathway and reducing neuroinflammatory reaction, or the targeted gene of Dracocephalum Moldavica L has not been cleared yet. Herein, our research considers miR-148a-3p as the new target of RAGE/α7nAChR signaling pathway and studies the genetic level of the interaction mechanisms between the molecules in inflammatory signaling pathways. We will comprehensively use the methods and techniques of bioinformatics, molecular biology and protein chip technology. And we will investigate the function and pathology of miR-148a-3p in three aspects of gene function, pathological pathways and drug reaction. Finally, we will get to the aim of exploration of miR-148a-3p cascades or its feedback multidirectional mechanisms in AD inflammatory injury through RAGE/α7nAChR signal pathway, and then explain the mechanisms of Dracocephalum Moldavica L of preventing and treating AD based on the neuroinflammatory mechanism.

非编码微小RNA的表观遗传及转录调控在AD复杂发病机制中发挥了重要作用。我们此前发现，AD小鼠脑内miR-148a-3p的表达降低与神经炎症密切相关，并受到靶基因RAGE及关联通路分子α7nAChR的反向特异性调控。近期又发现，中药香青兰提取物防治AD的作用与其活性单体调节miR-148a-3p/RAGE信号通路显著相关。但miR-148a-3p通过靶基因及关联信号途径减缓AD神经炎症的时序性与特异性机制及香青兰的干预位点均不明确。本课题拟以miR-148a-3p作为调节RAGE/α7nAChR通路的新靶标，对炎症信号分子间相互作用机制进行基因水平的追溯研究。综合利用生物信息学、分子生物学、蛋白通路芯片等技术，从基因功能、病理通路、药物作用等方面，探究miR-148a-3p经由RAGE/α7nAChR在AD炎症损伤中形成的串联、级联或反馈性的多向调控机制，阐释香青兰抗炎防治AD的作用机理。

传统的基于单一靶标或单一信号通路的药物发现策略对于机制复杂、病因不明的阿尔茨海默病（Alzheimer' disease，AD）而言，一直没有揭示该病的病理机制，也没有找到理想的防治药物。非编码RNA 和靶标基因-分子形成多途径信号参与了该病的进展，在此基础上寻找多靶标联合起效的药物，可能为防治该病提供有效手段。本项目拟以miR-148a-3p作为干预RAGE/α7 nAChR信号通路的调控基因进行确证研究，对α7 nAChR、RAGE等神经炎症关键分子的相互作用进行探讨研究，并对香青兰活性单体开展防治AD的机制研究。.本项目实施按照任务计划书有序进行，完成研究计划和成果指标。通过在不同病理进程的AD动物大脑基因芯片中挖掘数据信息和功能研究，首次明确了miR-148a-3p的异常变化在AD进程及神经炎症中的作用，阐释了miR-148a-3p特异性调节RAGE/α7 nAChR信号轴线的机制，确证了miR-148a-3p在神经炎症信号通路的靶标基因，揭示了miR-148a-3p/RAGE/α7 nAChR形成的网络信号在AD的病理作用，以及阐述了香青兰活性化合物基于miR-148a-3p/RAGE/α7 nAChR的抗AD作用特点和作用机制。上述研究工作在SCI收录期刊发表研究性论文9篇，获得国际专利1项，申请国内外专利3项，获得学术奖励2项，培养青年科技人才2人，培养博士生、硕士生研究生3人。.综上所述，本项目明确了miR-148a-3p信号轴线的抗AD机制，阐释了香青兰活性物质基于miR-148a-3p信号通路中基因表达及关键靶标变化的抗AD机理，为以RAGE和α7 nAChR为作用靶标、以miR-148a-3p表达调控为靶向的药物研究提供了理论基础和实验依据。