电针对阿尔茨海默病模型SD大鼠学习记忆功能及海马神经细胞凋亡的影响

Influence of electro acupuncture on learning and memory capability and hippocampal neuronal apoptosis in AD model SD rats

目的观察电针对阿尔茨海默病（AD）模型SD大鼠学习记忆、海马神经细胞凋亡、谷氨酸（Glu）含量、N-甲基-D-天门冬氨酸（NMDA）受体、caspase-3蛋白、自噬相关蛋白Beclin-1表达的影响。方法 48只健康雌性SD大鼠随机分为AD模型组、假手术组、正常对照组和电针组,每组12只。采用Aβ25~35建立AD模型,Morris水迷宫观察大鼠学习记忆能力,尼氏染色观察海马神经元数量,细胞术检测各组海马神经元的凋亡,高效液相色谱法检测海马Glu含量,免疫组化法检测海马组织NMDA受体表达,Western blotting检测海马caspase-3和beclin-1蛋白的表达。结果与AD模型组比较,电针组SD大鼠在第Ⅱ象限停留时间所占的百分比较大、跨越平台次数较多、目标象限停留时间较长、平台象限停留路程占总路程百分比较大、平均潜伏期较短,学习记忆能力有明显改善;海马区神经元凋亡率较低,NMDA受体功能增强,Glu含量、caspase-3和Beclin-1蛋白表达减少（P〈0.05）。结论电针治疗可改善AD大鼠学习记忆功能,抑制神经元凋亡,上调NMDA受体功能,减少凋亡相关蛋白的含量。

Objective To observe the influence of electro acupuncture on learning and mem- ory capability, neuronal apoptosis, the content of Glu, the expression of N- methyl- D- aspartate （NMDA） receptors, caspase-3 and autophagy-related protein Beclin-1 in Alzheimer＇s disease （AD） model SD rats. Methods A total of 48 healthy female rats were randomly and equally divided into AD model group, sham operation group, normal control group and electro acupuncture （EA） group. AD model rats were established with Aβ25～35 protein injected into bilateral hippocampus. Morris water maze was applied to investigate the learning and memory capability of rats. Nissl stain was used to count the number of hippocampal neurons. Flow cytometry was carried out to detect neu- ronal apoptosis. High performance liquid chromatography （HPLC） was applied to detect Glu con- tent in hippocampus. Immunohistoehemistry （IHC） was carried out to detect the expression of NM- DA-R in hippocampus. Western blotting was used to detect the expression of caspase-3 and beclin-1 in hippocampus. Results Compared with theAD model group, rats＇ abilities of learning and memory improved significantly （ P 〈 0.05） in the EA group, which manifested such as lowest percentage of time spending in the second quadrant and most time spending in the target quadrant, the high- est frequencies of leaping over platform and percentage of the distance of platform quadrant, and the shortest average latency. There was lower neuronal apoptosis proportion in hippocampus. EA increased hippocampus NMDA-R expression significantly （P 〈 0.05）, with decreased Glu, cas- pase-3, Beclin-1 protein expression correspondently. Conclusion EA therapy can improve learn- ing and memory capability in AD rats, inhibit the apoptosis of nerve cells, up-regulate the func- tion of NMDA-R and reduce the expression of apoptosis-related protein.