夹脊电针对ALS-SOD1^(G93A)转基因小鼠腰髓前角运动神经元形态学的影响

Effect of Electroacupuncture at Jiaji Points (EX-B2) on Morphology of Lumbar Spinal Anterior Horn Neurons in the Transgenic Mice with Amyotrophic Lateral Sclerosis (ALS)

目的:观察夹脊电针对ALS-SOD1^(G93A)转基因小鼠腰髓前角运动神经元形态学的影响。方法:本实验以ALS-SOD1^(G93A)转基因小鼠为实验对象,选用经PCR鉴定的ALS-SOD1^(G93A)转基因小鼠24只,按照随机数字表法将小鼠随机分为电针组、手针组和模型组(均n=8只),取同窝野生型小鼠8只作为阴性对照组。于小鼠日龄60天开始干预,电针组针刺双侧L_(1~2)、L_(5~6)夹脊穴,躯干同侧2 Hz电针治疗;手针组针刺双侧L_(1~2)、L_(5~6)夹脊穴;模型组予以捆绑固定,20 min/次,2次/周,共4周;阴性对照组不做任何处置。于小鼠日龄120天时取腰膨大组织进行HE染色和尼氏染色来观察各组小鼠腰髓前角运动神经元胞体及胞核的状态并对运动神经元进行计数,透射电镜观察腰髓前角神经元超微结构变化。结果:HE染色可见模型组小鼠腰髓前角运动神经元数目明显减少,神经细胞结构不清、固缩,胞质胞核染色不清,核固缩,出现空泡状改变;与模型组比较,手针组和电针组小鼠腰髓前角运动神经元数目增多,神经细胞结构形态有所改善,空泡化减少,且电针组效果优于手针组。尼氏染色可见模型组小鼠腰髓前角运动神经元数目明显减少,非神经细胞增多,病变的神经元出现尼氏体不清,核固缩,细胞体积减小;与模型组比较,手针组和电针组小鼠腰髓前角运动神经元数目增多,神经细胞结构形态有所改善,能够明显减轻运动神经元的丢失,且电针组效果优于手针组。运动神经元计数:模型组小鼠腰髓前角运动神经元数目明显减少(P<0.01);手针组和电针组小鼠腰髓前角运动神经元数目较模型组增多(P<0.05或P<0.01),且电针组较手针组增多更明显(P<0.05)。透射电镜可见模型组小鼠腰髓前角神经细胞萎缩,细胞膜皱缩,线粒体不同程度肿胀,嵴出现断裂、减少,有些线粒体虽可辨认,但已呈空泡状,核膜凹陷变形或部分破裂,异染色质聚集成块,变性的神经细胞周围可见胶质细胞围绕形成卫星现象;与模型组比较,手针组和电针组小鼠腰髓前角神经细胞超微结构有所改善,且电针组改善更明显。结论:夹脊电针能够改善ALS-SOD1^(G93A)转基因小鼠腰髓前角运动神经元形态学变化,其效果优于手针治疗。

Objective：To observe the effect of electroacupuncture at Jiaji point （EX -B2） on morphology of lumbar spinal anterior horn neurons in the transgenic mice with ALS. Methods ：24 ALS -SOD1^G93A transgenic mice were randomly divided into three even groups, that is electroacupuncture （group E）, acupuncture group （group A ） and model group （group M ）. Eight non -transgenic littermates were assigned to blank group （group B）. Interventions to ALS - SOD1^G93A transgenic mice began from their age of 60 days. Group E was treated by electroacupuncture at Jiaji points （EX - B2 ）, with the electrical stimulation at the frequency of 2 Hz. The four - week intervention was 20 minutes per time and twice a week. Group A was only treated by acupuncture at Jiaji points （EX -B2 ） and group C was disposed by firm binding. None intervention was given to group B. At the age of 120 days, HE staining, Nissl staining and transmission electron microscopy were used to ALS - SOD1^G93A transgenic mice to observe the anterior horn of the lumbar spinal cord pathological morphology, and detect the amount of motor neurons. Results：Analysis from HE staining, Nissl staining and transmission electron microscopy showed that group E and group A could significantly reduce morphological lesions and increase the amount of motor neurons compared with group C （ P 〈 0.05 or P 〈 0.01 ）, and group E was better than group C. Conclusion ：Electroacupuncture at Jiaji points（ EX -B2 ） can improve morphological changes of motor neurons in the anterior horn of the lumbar spinal cord in ALS -SODI G93A transgenic mice, which is better than by manual acupuncture.