B淋巴细胞在电针改善中风后迟发性认知功能损害中的作用研究

Delayed Cognitive Impairment following Stroke is showing a strong impact on the quality of life of post stroke patients and increasing the financial burden of the society. But the mechanism is still unclear and there are no effective treatments. We found that electroacupuncture(EA) treatment could alleviate delayed Cognitive Impairment in mouse model with distal middle cerebral artery occlusion (DMCAO). A recent research indicated that the delayed Cognitive Impairment in DMCAO mouse model was mediated by the activated B lymphocytes which infiltrated into the cortex post stroke. Besides, EA could activate the cholinergic nerve system which could regulate the activation of B lymphocytes. Here we hypothesize that through activating cholinergic nerve system, EA inhibits the activation and infiltration of B cells into cerebral cortex, decreases the secretion of autoantibodies, reduce the hippocampus injury, and alleviate delayed Cognitive Impairment. In this project, we will use B cell adoptive transfer and cholinergic nerve system interference and then determine their effects on LTP, spine, mouse behavior to verify the key role of B cells in mediating the alleviating effects of EA on delayed Cognitive Impairment in DMCAO mouse model. We are trying to further explain the mechanism of how EA alleviates delayed cognitive Impairment post stroke from the point of neuro-immune modulation.

中风后迟发性认知功能损害严重影响患者生活质量、增加社会经济负担，其发生机制尚不明确且无有效防治手段。我们的预实验发现中风后电针治疗可以减轻小鼠大脑中动脉远端阻塞（DMCAO）模型的迟发性认知功能损害，且最新研究发现DMCAO小鼠的迟发性认知功能损害与B细胞的激活和中枢浸润有关，已知电针可以激活胆碱能神经系统，且B细胞的激活受胆碱能系统调控。由此，我们提出假说：电针通过激活中枢和（或）外周的胆碱能神经系统，抑制中风后B淋巴细胞的激活和中枢浸润，减少自身抗体的分泌，从而减轻其对海马神经元的损伤，改善中风后迟发性认知功能损害。本项目拟采用B细胞的过继转移，中枢及外周胆碱能神经系统的干预等方法，并通过行为学，电生理，形态学，生物化学等检测手段明确B淋巴细胞在电针改善中风后迟发性认知功能损害中的关键作用，从神经免疫调节的角度深入阐释电针防治中风后迟发性认知功能损害的病理生理机制。

背景：中风后迟发性认知功能损害严重影响中风后患者工生活质量、增加社会经济负担，其发生机制尚不明确且无有效防治手段。大脑中动脉远端缺血（DMCAO）小鼠是研究中风后迟发性认知功能损害的合适动物模型。.方法及结果：我们对模型小鼠进行电针百会及足三里穴位刺激7周，通过物体位置识别（OLT）及Y迷宫进行行为学检测，发现DMCAO小鼠迟发性认知功能损害得到改善。通过高尔基染色，发现海马神经元树突棘密度降低得到改善，突触蛋白PSD95及SYP表达下降也得到改善，同时免疫印迹显示B淋巴细胞中枢浸润得到抑制。进一步，通过B淋巴细胞脑定位注射，增加B淋巴细胞中枢浸润，发现此干预削弱了电针改善DMCAO小鼠迟发性认知功能损害的作用。膈下迷走神经切断使得电针抑制B淋巴细胞中枢浸润的作用削弱，同时电针改善DMCAO小鼠迟发性认知功能损害的作用也被削弱。.结论：电针通过迷走神经抑制DMCAO小鼠B淋巴细胞的中枢浸润，减轻自身免疫对海马神经元的损伤，改善中风后迟发性认知功能损害。本研究从神经免疫调节的角度深入阐释了电针防治中风后迟发性认知功能损害的病理生理机制。