基于布鲁斯效应探讨益气活血方药改善学习记忆的神经机制

Learning and memory is a hot issue of neural science and geratology at present, yet no study well elucidated its complex neural mechanism so far. If memory signal could be magnified enough to form biological function, memory process would be authentically reflected and easily verified. Pheromonal memory is a typical sample of such kind of studies. Bruce Effect may appropriately verify whether female mouse has formed memory induced by pheromone from spouse male mouse, which is also used as a standard for judging single-trial memory formation. We have demonstrated that WNK, a Chinese herbal formula established according to tonifying qi and promoting blood circulation, could improve learning and memory significantly in multiple model animals and enhance PKC activity. We also initially found that PKCα plays a key role in AOB synaptic plasticity. According to Chinese Medicine theory and previous results, based on Bruce Effect, we are to study 1) the mediating role of PKCα in pregnancy, AOB synaptic plasticity and pheromonal memory formation in Balb/c and PKCα-knock-out mice; 2) the candidate effects and neural mechanisms of WNK on modulating single-trial memory through activating PKCα pathway. With interdisciplinary approach, our work will profoundly elucidate the scientific contents of totifying qi and promoting blood circulation therapy for strengthening brain function and enhancing intelligence, and provide new insights and experimental evidence for studies on learning and memory.

学习记忆是当今神经科学和老年医学的热点问题之一，但现有研究不足以揭示其复杂的神经机制。若记忆信号被放大成生理功能，可更真实地反映记忆过程并容易获得验证，信息素记忆是此类研究的典型实例。布鲁斯效应可恰当地验证雌鼠是否对配偶雄鼠信息素产生了记忆，是单次试验记忆形成的检验标准。课题组已证实益气活血方药维脑康（WNK）可明显改善多种模型动物的学习记忆、显著增强PKC活性，并首次发现PKCα对AOB突触修饰起决定作用。根据中医理论和前期工作，借助布鲁斯效应，本项目以正常Balb/c小鼠和PKCα基因敲除小鼠为对象，研究1）PKCα对妊娠发育、AOB突触可塑性、信息素记忆形成的介导作用；2）WNK通过活化PKCα通路调节单次试验记忆形成的作用和神经机制。本项目采用多学科交叉的方法，揭示益气活血法健脑益智的科学内涵，并为学习记忆研究提供全新的视角和方法。

学习记忆是当今神经科学的热点问题之一，但现有研究不足以揭示其复杂机制。若记忆信号被放大成生理功能，可更真实地反映记忆过程并易获验证。布鲁斯效应可恰当验证雌鼠是否对配偶雄鼠信息素产生记忆，是判断单次试验记忆形成的公认标准。.妊娠终止研究结果提示，气味刺激48h后，正常小鼠子宫内膜明显变厚，细胞间隙增大，局部可见妊娠间质反应；PMA组子宫内膜COX2、LIF、HOXA-10表达升高，che组LIF、HOXA-10表达降低。与其他时相比较，气味刺激后15d小鼠流产率、卵巢指数、子宫指数显著升高，妊娠小鼠子宫壁内膜非常显著性增厚，腺体明显增多，可见妊娠间质反应，孕激素水平较非妊娠和妊娠终止小鼠显著升高；NE、Glu水平显著下降；che组PKC、PKCɑ表达水平显著降低；小鼠整体表现为雄鼠气味刺激后胚胎数目减少，陌生雄鼠较配偶影响更为明显；副嗅球组织学改变明显，如颗粒及僧帽细胞带长明显缩短，颗粒及僧帽细胞、小球及小球周细胞明显减少。.药效学结果提示，无气味刺激时，WNK药后流产率、血清孕激素水平显著降低，雌激素水平显著升高。气味刺激后15d，药后妊娠小鼠子宫内膜较厚、间质较疏松，COX2、LIF、HOXA-10表达明显升高，集中在腔上皮、腺上皮及血管内皮细胞胞浆；CA1区神经元线粒体和突触明显减少，部分胞内可见炎性反应；副嗅球部分僧帽、刷状及颗粒细胞线粒体受损，但较PMA、che组超微结构完整、线粒体多；配偶组颗粒及僧帽细胞较多，GAP-43、mGluR2表达显著升高；陌生雄鼠组NE水平及SOM、GAP-43、α1-AR表达显著下降。.本研究提示，PKCɑ通路可能参与了妊娠终止反应，益气活血中药可能干预AOB突触可塑性、影响信息素记忆形成，其作用与PKCɑ通路密切相关，进一步阐明益气活血中药改善学习记忆的可能神经机制，为学习记忆研究提供了新的思路和方法。.本项目培养博、硕士研究生毕业各1名，博士研究生在读1名；主编/参编出版专著3部，发表核心期刊、会议论文各1篇，已投稿核心期刊论文1篇、SCI论文3篇，其中2篇IF>4（均已修回）、1篇IF>3（已投出）；申请专利2项；获省部级科学技术一等奖1项。