基于miR-206对BDNF的调控探讨循经按摩对脑梗死大鼠运动功能重建的机制

Cerebral infarction is a common and frequent disease of the nervous system,high morbidity,easily resulting in motor, sensory and cognitive dysfunction.Meridian massage showed better efficacy,but its mechanism remains to be further defined.Based on the literature and preliminary studies,the following hypothesis was presented:the role of meridian massage can work on muscle tissue, by regulating the expression of miR-206, further promote muscle BDNF expression,thereby contribute to functional changes of synapses, finally lead to rebuilding damaged nerve function. Therefore,the research group intended to treat rats model of cerebral ischemia by using meridian massage, which effect would be observed on movement function recovery, and on neuronal cell ultrastructure in ischemic zone, as well as on skeletal muscle miR-206 and BDNF.In addition,combined with molecular biology techniques,the effect on BDNF targeting at enhancement or inhibition the expression of C2C12 myoblasts miR-206 also was observed.This research investigated the regulation of skeletal muscle miR-206 mediating BDNF in the meridian massage, and revealed the mechanism of rehabilitation in promoting the recovery of movement function after cerebral infarction.

脑梗死是神经系统的常见病和多发病，致残率高。遗留运动、感觉、认知等功能障碍。循经按摩治疗本病具有较好的疗效，但其机制有待进一步明确。基于文献及前期研究，提出：循经按摩作用于肌肉组织，通过调节肌肉组织miR-206表达，进而促进肌肉BDNF表达，促进神经肌肉接头突触传递效率，促进运动神经元的存活，重建受损神经，改善运动功能的假说。为此，课题组拟运用循经按摩治疗脑缺血大鼠模型，观察其对脑梗死大鼠运动功能恢复、缺血区神经细胞超微结构及骨骼肌miR-206、BDNF的影响，并结合分子生物学技术，靶向增强、抑制C2C12成肌细胞miR-206表达，观察其对BDNF的影响。探讨骨骼肌miR-206调控BDNF在循经按摩中的作用，揭示循经按摩促进脑梗死后运动功能重建的机制。

项目研究内容已完成.第一方面：动物实验：制备MACO大鼠模型，对MCAO大鼠进行循经按摩后，应用Z-longa量表对大鼠神经功能进行评分、HE染色、超微电镜、PCR、ELISA、WB方法分别检测大鼠的脑组织、全血、腓肠肌组织。结果证实大鼠神经功能评分得到改善；脑组织超微结构显示循经按摩后神经细胞排列较模型组整齐且排列紧密，且随着治疗时间延长逐渐变得整齐且排列紧密，循经按摩组相对模型组自噬情况较轻微；腓肠肌、全血miR-206循经按摩组较模型组降低，全血及腓肠肌的BDNF mRNA循经按摩组较模型组增加。全血及腓肠肌的BDNF蛋白含量循经按摩组较模型组增加。循经按摩可以改善脑卒中大鼠的神经运动功能，可能与促进BDNF的表达有关，可能是通过调节miR-206表达，进而促进肌肉BDNF表达，为循经按摩治疗脑卒中患者功能障碍提供理论依据。.第二方面：细胞实验：分为miR-206过表达组（LV-hsa- miR -206(1569-11）慢病毒包装靶向增强 miR-206的shRNA载体）及miR-206抑制表达组（LV-hsa-miR-206-inhibition慢病毒包装靶向抑制miR-206的shRNA 载体），每组又分为0%，5%，10%，15%，20%/10cycles/min的不同牵张应力组，PCR、WB检测miR-206及BDNF的mRNA及蛋白表达。结果显示10%牵张应力作用下C2C12成肌细胞BDNF mRNA表达与对照组相比显著升高，15%牵张应力作用下miR-206 mRNA与对照组比较显著降低；牵张应力组为10%时BDNF蛋白的表达上升到最大值。