针康法调控cIAP1降低脑缺血后神经血管损伤的机制研究

The clinical effect of acupuncture-rehabilitation method is precise and wide applied in stroke rehabilitation,but the mechanism has not been elucidated. Acupuncture-rehabilitation method can regulate nerve vascular unit reconstruction after cerebral ischemia,protecting neurons/vessels in preliminary studies.Whether there is a shared molecule underlying acupuncture-rehabilitation method induce protection in neurons and vascular endothelial cells remains unknown? The neurons and vascular endothelial cell apoptosis will be observed after acupuncture-rehabilitation method situation in cerebral ischemia animal model in this study;CIAP1 silent in brain,the changes of neurons and microvessels and their molecular markers will be observed after acupuncture-rehabilitation method situation by Nissl staining, electron microscopy and immunoblotting methods in each experimental groups.The oxygen glucose deprivation (OGD) models in neurons and vascular endothelial cells are made in vitro. cIAP1 genes silent by the lentiviral vector system to mediate and BDNF gived to analog acupuncture-rehabilitation method effect in vitro, the changes of neurons and microvessels and their molecular markers will be observed. The hypothesis that"cIAP1 is a shared molecule underlying acupuncture-rehabilitation method induced protection in neurons and vascular endothelial cells against cerebral ischemia " will be proved in vivo and vitro, providing the theory basis for acupuncture-rehabilitation method applied in clinical.

针康法作为脑卒中康复常用方法，临床疗效确切，其作用机制尚不十分清楚。我们前期研究发现，针康法能促进脑缺血后神经血管单元重构，对神经元/血管都起到保护作用。是否存在某共同靶点介导针康法对神经元、血管的保护作用，是本课题关注的问题。本项目拟在脑缺血动物模型上，观察针康法干预后神经元、血管内皮细胞凋亡的情况；在体沉默脑内cIAP1，采用尼氏染色、电镜、免疫印记等方法观察针康法干预后各实验组神经元、微血管及其分子标志物的变化；体外建立神经元、血管内皮细胞氧糖剥夺（OGD）模型，沉默cIAP1基因，给予BDNF处理模拟针康法的体外效应，观察各实验组细胞损伤情况及神经血管分子标志物的变化。从体内外两方面验证"cIAP1是针康法诱导神经元保护、血管保护的共同靶点"这一假说，为针康法临床推广应用提供理论依据。

针康法作为脑卒中康复常用方法，临床疗效确切，其作用机制尚不十分清楚。我们前期研究发现，针康法能促进脑缺血后神经血管单元重构，对神经元/血管都起到保护作用。本项目拟在脑缺血动物模型上，观察针康法干预后神经元、血管内皮细胞凋亡的情况；从体内、体外的角度沉默脑内cIAP1，采用尼氏染色、电镜、免疫印记等方法观察针康法干预后各实验组神经元、微血管及其分子标志物的变化，研究发现：针康法可以增加脑缺血大鼠前肢抓握力量，提高运动协调能力，促进运动功能重建；针康法可以降低脑缺血大鼠缺血半暗区神经元凋亡，减少细胞凋亡数量，发挥神经保护作用；神经元上cIAP1、cIAP2、XIAP、Survivin表达与神经元存活密切相关，脑缺血后cIAP1表达下调，存活神经元减少，针康法干预后cIAP1表达上调，存活神经元增加。针康法通过调节cIAP1表达，抑制caspase-9、caspase-8和caspase-3活化，降低脑缺血后神经元、血管内皮细胞凋亡，减少细胞凋亡数量。阐明cIAP1参与脑缺血后神经血管重塑，是针康法诱导神经元保护、血管保护的共同靶点；项目资助发表论文15篇，获得中国康复医学会科学技术一等奖1项，获得1项发明专利、3项实用新型专利。2人晋升副教授，1人评为省级龙江英才。培养博士研究生3名，硕士研究生1名。该项目的完成为针康法的临床应用提供坚实的理论指导。