针刺通过Orexin A调控昼夜节律Clock/Bmal1- Pers/Crys反馈环路改善血管性轻度认知障碍的作用机制

Vascular mild cognitive impairment (VMCI) is the early stage of various types of dementia. Moreover, post stroke sleep deprivation (SD) is an important factor leading to its occurrence and development. It is of great practical significance to give full play to the advantage of "preventive treatment of disease" in TCM for improving insomnia and amnesia after stroke. Our research group supposed the core pathological of VMCI is due to over-expression of orexin A, which lead to the mis-expression of read-out gene Bace1 in circadian rhythm feedback loop Clock/Bmal1-Pers/Crys, finally triggered Aβ accumulation. Acupuncture is an effective treatment on VMCI, which is related to the regulation of circadian rhythm and underlying pathway. From the perspective of reducing orexin A level and the Aβ accumulation which Clock/ Bmal1-Pers/Crys feedback loop participated, this research intends to dissect 2 scientific questions: 1. The mechanism of the SD effect on VMCI after stroke. 2. Utilizing qRT-PCR, Western blot and other techniques performed in the rat model to explore the mechanism of acupuncture on preventing VMCI and reversing its conversion into dementia. Gene silencing technique (RNAi) was used for investigating potential acupuncture targets. The implementation of this project will provide a candidate strategy for clinical prevention and treatment of VMCI, and reveal the theoretical implication of insomnia and amnesia after stroke from the aspect of TCM.

血管性轻度认知障碍（VMCI）是多种痴呆的前期阶段，而卒中后睡眠剥夺（SD）是导致其发生发展的重要因素，发挥中医“治未病”优势对于改善卒中后“失眠”-“健忘”具有重大现实意义。课题组认为食欲素A（Orexin A）水平升高，导致昼夜节律Clock/Bmal1-Pers/Crys反馈环路输出基因Bace1过表达，引发β-淀粉样蛋白（Aβ）沉积是VMCI发生的核心病理环节。针刺可以有效的防治VMCI，该作用与调节昼夜节律及通路有关。课题组拟从降低Orexin A水平及Clock/Bmal1-Pers/Crys反馈环路导致的Aβ沉积角度探讨：（1）卒中后SD对VMCI影响的机制；（2）采用动物模型及后续qRT-PCR、WB等技术评价针刺防治VMCI、逆转其向痴呆转化的机制；采用RNAi技术探讨针刺作用靶点。本课题的实施将为临床防治VMCI提供新策略，揭示卒中后“失眠”-“健忘”的中医学理论内涵。

本项目针对中医临床有效、西药难以解决的认知功能障碍的关键科学问题，以临床广泛应用的针灸作为干预手段，从整体水平着手，采用分子生物学等理念、方法和关键技术，揭示针刺干预认知障碍疾病中的效应特点，阐明针刺疗法治疗可以有效降低兴奋性神经递质OrexinA水平，增强突触可塑性，抑制海马神经元细胞焦亡等多种复杂作用机制，彰显传统中医药治疗特色，最终为促进针刺作用的科学表达以确保临床精准治疗提供研究范例。具体成果体现在3方面：1. 发现电针通过调节Orexin A介导的cAMP/PKA/CREB信号通路改善衰老相关认知障碍的神经可塑性；2. 发现电针通过Orexin A调控昼夜节律Clock/Bmal1-Pers/Crys 反馈环路抑制Aβ的产生发挥改善VMCI的认知功能；3. 发现电针通过NLRP3/Caspase-1信号通路抑制海马神经元焦亡改善衰老相关认知障碍的认知功能。本项目组共发表相关学术论文4篇，其中SCI论文3篇，中文科技核心期刊1篇。单篇最高影响因子文章已在Oxidative Medicine and Celluar Longevity（IF：7.31）发表，单篇最高他引6次。培养黑龙江省优秀青年基金获得者1人，获得国家发明专利1项。