目的 探讨意向性运动对局灶性脑缺血大鼠行为学及缺血周围脑组织细胞外信号调节激酶(ERK)-环磷酸腺苷反应元件结合蛋白(CREB)通路表达的影响.方法 选取清洁级健康雄性SD大鼠,采用线栓法制备大脑中动脉栓塞(MCAO)模型144只,按随机数字表法分为模型组、游泳运动组、饲养环境改变组和意向性运动组,每组36只大鼠;各组大鼠按术后时间分为脑缺血7d、15d和30 d三个观察时间点,每个观察时间点各12只大鼠,动态观察各组大鼠每个观察时间点的行为学变化,并采用逆转录-聚合酶链反应(RT-PCR)和免疫荧光染色检测缺血周围脑组织ERK-CREB通路的表达水平.结果 意向性运动组各时间点爬梯频率明显高于环境改变组(P<0.05);各组大鼠神经功能缺损评分于缺血再灌注3d后随时间呈下降趋势,意向性运动组的神经功能缺损评分(1.33 ±0.49、0.50 ±0.52、0.08 ±0.29)较模型组(2.00±0.60、1.58±0.67、1.00 ±0.60)、游泳运动组(2.50 ±0.52、1.00 ±0.60、0.50 ±0.52)及环境改变组(1.92 ±0.52、1.00±0.43、0.58±0.52)均明显降低(P<0.05);再灌注7、15和30 d,意向性运动组缺血周围脑组织ERK、CREBmRNA及pERK、pCREB蛋白的表达与其他三组比较均明显增加(P<0.05).结论 意向性运动能改善局灶性脑缺血后的神经功能,可能与其上调并激活缺血周围脑组织ERK-CREB通路有关.
Objective To explore the effects of willed movement on neurological performance and the extracellular signal-regulated kinase (ERK) and cAMP response element binding protein (CREB) pathway in rats following focal cerebral ischemia.Methods Reversible middle cerebral artery occlusion was induced in 144 male SpragueDawley rats using intraluminal sutures,and they were randomly divided into a control group,a swimming exercise group,an environment modification group,and a willed movement group.The observation time points were at 7,15 and 30 days after reperfusion.A behavioral test was performed to evaluate any neurological deficiency.Reverse transcription PCR (RT-PCR) and immunofluorescence were used to detect the ERK and CREB responses in terms of mRNA and phosphorylated ERK (pERK) and phosphorylated CREB (pCREB) protein in the peri-ischemic brain tissue.Results The climbing frequency of the willed movement group was significantly higher than that of the environment modification group.Three days after reperfusion the neurological deficit scores of all groups began to decrease,and that of the willed movement group had decreased significantly more than in the other three groups at all time points.ERK/CREB mRNA and pERK and pCREB protein expression were dramatically up-regulated in the willed movement group at 7,15 and 30 days after reperfusion,significantly more than in the other three groups.Conclusions Willed movement may promote motor recovery by up-regulating and activating the ERK/CREB pathway following focal cerebral ischemia.