基于lncRNA 探讨针刺抗MCAO大鼠脑缺血后神经血管单元损伤的作用机制研究

The pathological mechanism of ischemic stroke has not been fully elucidated. The.most disporting is the failure of clinical trials of candidate drugs designed by targeting one of the pathological injury mechanisms．So the search for new therapeutic targets is the key to improve the clinical efficacy. The role of lncRNA in tumor, neurodegenerative disease have attracted much attention, and the study achieved some results. Comparing with tumor and neurodegenerative disease，study of IncRNAs in post-ischcmia is very limited. Previous studies show that there is good curative effect of acupuncture in treating cerebral ischemia..This study Use the Rat IncRNA microarray to observe the IncRNAs expression profile of ischemia rat brains and to evaluate the therapeutic effects of acupuncture in the Middle cerebral artery occlusion model(MCAO) in rats, Use bioinformatics techniques(KEGG pathway analysis and GO analysis) analyze the microarray results，construct the lncRNAs-mRNAs co-expression network with Weighted Correlation Network Analysis , and use RT-PCR to verify the differential expression 4-6 lncRNAs >2-5 folds, to find the possible lncRNAs which may play the roles in multi-pathological pathway, and Regulative action of acupuncture on it.From the cell biological regulatory networks we will thoroughly explanate the mechanisms of brain injury after cerebral ischemia, the material basis and effective target of acupuncture protecting neurovascular unit(NVU). This study will help to reveal the mechanism of acupuncture for treatment of cerebral ischemia, and provide objectively theoretical and experimental basis for acupuncture treatment of cerebral ischemia.

缺血性脑卒中病理机制尚未完全阐明。针对脑缺血病理损伤中的某个靶点开发的治疗候选药物的临床疗效均不理想，寻找新的治疗靶点是提高临床疗效的关键。lncRNAs在肿瘤、神经退行性疾病中的作用引起高度关注，并取得了一定成果，但在缺血性脑卒中的研究相对较少。课题组前期研究表明针刺治疗脑缺血有较好的疗效。本研究拟采用高通量测序方法检测MCAO模型大鼠脑组织lncRNAs的表达及其针刺对其干预效应，运用GO分析、KEGG分析等方法对lncRNA的表达谱进行分析，通过权重基因共表达网络分析法构建差异基因的lncRNAs-mRNAs共表达网络，并运用RT-PCR技术对差异表达2-5倍以上的4-6个lncRNAs进行验证，从细胞生物学调控网络角度深入阐释脑缺血后脑损伤发生的病理机制及针刺保护NVU的可能物质基础及有效作用靶点。本研究将有助于揭示针灸治疗脑缺血的效应机制，为针刺治疗脑缺血提供客观的实验与理论依据

缺血性脑卒中病理机制尚未完全阐明。针对脑缺血病理损伤中的某个靶点开发的治疗候选药物的临床疗效均不理想，寻找新的治疗靶点是提高临床疗效的关键。lncRNAs在肿瘤、神经退行性疾病中的作用引起高度关注，并取得了一定成果，但在缺血性脑卒中的研究相对较少。.本研究采用高通量测序方法检测MCAO模型大鼠脑组织lncRNAs的表达及其针刺对其干预效应，运用GO分析、KEGG分析等方法对lncRNA的表达谱进行分析，通过权重基因共表达网络分析法构建差异基因的lncRNAs-mRNAs共表达网络，并运用RT-PCR技术对差异表达的lncRNAs进行验证，研究脑缺血后脑损伤发生的病理机制及针刺保护NVU的可能物质基础及有效作用靶点。.在本实验中将针刺的时间点分设脑卒后5分钟，30分钟和60分钟结果发现,三个时间点的针刺作用能有效减少脑梗死体积，且三组之间并没有统计学意义，但在脑卒后30分钟进行电针治疗可以有效地减少大鼠的死亡率。电针水沟穴可以减少MCAO大鼠的脑梗死体积，降低大鼠神经行为学评分。提高MCAO大鼠缺血半暗带的CD34含量，促进缺血半暗带微血管新生。降低MCAO大鼠缺血半暗带的AQP4含量，进而影响大鼠患侧大脑的血脑屏障和脑水肿。改变MCAO大鼠缺血半暗带的细胞内结构形态和细胞器的功能，进而影响脑细胞的功能。研究结果表明lncRNA和mRNA在本次的实验中确实出现了差异的表达，针灸具有一定的调整作用。.本研究将有助于揭示针灸治疗脑缺血的效应机制，为针刺治疗脑缺血提供客观的实验与理论依据。