Effects of electroacupuncture at Fengchi(GB20)on motor function and GFAP/NeuN expression around the ischemic tissue of the motor cortex in MCAO rats

电针风池对MCAO大鼠运动功能及运动皮层缺血周围组织GFAP/NeuN表达的影响

Objective:To investigate the potential mechanism of electroacupuncture(EA)at bilateral Fengchi(GB20)in treating cerebral ischemia-reperfusion injury and to provide a scientific basis for future experimental research and clinical applications.Methods:Forty male specific-pathogen-free Sprague-Dawley rats were randomly divided into four groups:a normal group,a normal with EA group,a model group,and a model with EA group,with 10 rats in each group.The normal group received no intervention.The normal with EA group received EA at bilateral Fengchi(GB20).The model group underwent middle cerebral artery occlusion(MCAO)using the suture.The model with EA group underwent MCAO and received EA at bilateral Fengchi(GB20).Cerebral blood flow was monitored using a laser Doppler cerebral blood flow meter.Neurologic damage was assessed using the neurologic deficit score,and motor ability was observed using the CatWalk gait system.The expression of glial fibrillary acidic protein(GFAP)and neuronal nuclei(NeuN)protein,the neuron markers,was detected by Western blotting.The protein expression levels of GFAP and NeuN,as well as the number of positive cells in the motor cortex,were detected using immunofluorescence.Results:Compared to the normal group,the cerebral blood flow values in the model group and the model with EA group decreased by more than 50%during the modeling process(P<0.01)and returned to pre-modeling levels after reperfusion(P>0.05).The neurologic deficit score increased(P<0.05),the average motor velocity decreased(P<0.05),GFAP protein expression and the number of positive cells in the motor cortex increased(P<0.05),and the NeuN protein expression and the number of positive cells decreased(P<0.05)in the model group.Compared to the model group,the neurologic deficit score decreased(P<0.05),the average motor velocity accelerated(P<0.05),GFAP and NeuN protein expression and the number of positive cells in the motor cortex increased(P<0.01)in the model with EA group.Conclusion:EA at bilateral Fengchi(GB20)can reduce neuronal loss and increase GFAP and NeuN protein expression in the motor cortex of rats after ischemia-reperfusion,improve the motor function after ischemic stroke,and accelerate the recovery of balance and stability of the affected limbs.

目的:探讨电针双侧风池治疗脑缺血再灌注损伤的可能机制,为后续的实验研究和临床应用提供科学依据。方法:将40只雄性无特定病原体级Sprague-Dawley大鼠按照随机、对照原则分为空白组、空白电针组、模型组和模型电针组,每组10只。空白组不进行任何干预;空白电针组进行电针双侧风池;模型组进行线栓法大脑中动脉栓塞(MCAO)模型制备;模型电针组进行线栓法MCAO模型制备和电针双侧风池。采用激光多普勒脑血流仪监测脑血流变化;采用神经缺损评分评定神经损伤程度;使用CatWalk步态系统观察大鼠运动能力;免疫印迹法检测胶质纤维酸性蛋白(GFAP)及神经细胞标志物神经元核抗原(NeuN)蛋白表达;免疫荧光检测运动皮层GFAP、NeuN蛋白表达和阳性细胞数。结果:与空白组相比,模型组与模型电针组大鼠脑血流值在造模过程中下降达50%以上(P<0.01),再灌注后基本恢复造模前水平(P>0.05);模型组大鼠神经缺损评分升高(P<0.05),平均运动速度下降(P<0.05),运动皮层GFAP蛋白表达和阳性细胞数增多(P<0.05),NeuN蛋白表达和阳性细胞数减少(P<0.05)。与模型组相比,模型电针组神经缺损评分降低(P<0.05),平均运动速度增加(P<0.05),运动皮层GFAP、NeuN蛋白表达和阳性细胞数均增多(P<0.01)。结论:电针双侧风池能减少缺血再灌注后大鼠运动皮层神经元的丢失,增加运动皮层GFAP、NeuN蛋白的表达,改善缺血性中风后运动功能,加速患肢平衡与稳定性的恢复。