针康法对慢性脑低灌注大鼠海马形态结构及GAP-43蛋白的影响

Effect of Acupuncture-Rehabilitation Therapy on Hippocampal Morphology and GAP-43 Protein in Rats with CCH

目的:通过观察针康法对慢性脑低灌注(CCH)大鼠空间记忆和学习功能的影响,海马区形态结构以及生长相关蛋白-43(GAP-43)的表达变化,探讨针康法治疗慢性脑低灌注引起认知功能障碍的分子作用机制。方法:取60只雄性SD大鼠,随机选取12只设为假手术组,其余48只建立CCH模型,将建模成功的48只大鼠随机分为模型组、针刺组、康复组和针康组,每组12只。各组大鼠按照相应的治疗方案进行治疗。治疗4周,采用Morris水迷宫检测大鼠的行为学变化;HE染色观察海马区形态结构和神经元细胞的损伤情况;蛋白质免疫印迹(Western Blot)技术检测大鼠海马区GAP-43蛋白表达。结果:与模型组相比,各治疗组大鼠逃避潜伏期缩短,搜索原平台次数增加,海马细胞间排列密集齐整,细胞和突触结构较完整,突触细胞数量及神经元的丢失有所缓解,GAP-43蛋白表达水平上调,其中针康组的改善程度最为明显,差异有统计学意义(P<0.05)。结论:针康法能够显著改善慢性低灌注所致的认知功能减退,且疗效优于单纯的针刺治疗或康复训练,这可能与上调大鼠海马区GAP-43蛋白水平表达有关。

Objective:To explore molecular mechanism of acupuncture-rehabilitation therapy in treatment of cognitive dysfunction caused by chronic cerebral hypoperfusion(CCH),through observing effects of acupuncture-rehabilitation therapy on spatial memory and learning function,morphological structure of hippocampus and expression of growth associated protein-43(GAP-43)in rats with CCH.Methods:60 male SD rats were randomly divided into the sham operation group(n=48)and the CCH model group(n=48).The 48 successful modeling rats were randomly divided into the model group,the acupuncture group,the rehabilitation group and the acupuncture-rehabilitation group,with 12 rats in each group.The rats in each group were treated according to the corresponding treatment plan.After 4 weeks of treatment,the behavioral changes of the rats were detected by Morris water maze,the morphological structure of hippocampus and the damage of neurons were observed by HE staining,and the protein expression of GAP-43 in hippocampus was detected by Western Blot.Results:Compared with those in the model group,the escape latency of rats was shortened,the times of searching for the original platform was increased,the hippocampal cells were arranged densely and neatly,the structures of cells and synapses were more complete,the number of synaptic cells and neurons were reduced,and the protein expression of GAP-43 was up-regulated in other treatment groups,among which the improvements in the acupuncture-rehabilitation group were most significant(P<0.05).Conclusion:Acupuncture-rehabilitation therapy can significantly improve cognitive impairment caused by chronic hypoperfusion,and the curative effect is better than that of single acupuncture or single rehabilitation training,which may be related to up-regulation of GAP-43 protein expression in hippocampus of rats.