“通督启神”法两种电针对APP/PS1小鼠额叶皮层小胶质细胞TNF-α表达的影响

Effects of Two types of Electro-acupuncture Treatments Based on "Tongdu Qishen" Theory on the Expressions of Microglia and TNF-α in Frontal Cortex in APP/PS1 Double Transgenic Mice

目的:观察"通督启神"法不同电针对APP/PS1小鼠学习记忆能力及额叶皮层MG与TNF-α表达的影响,探讨不同电针治疗AD的疗效差异及其作用机制。方法:APP/PS1小鼠随机分为模型组、脉冲电针组和音乐电针组,C57BL/6小鼠为正常对照组,每组8只。先取"人中"点刺,后取"百会"、"印堂"二穴,连接脉冲电针及音乐电针,留针20 min。正常对照组与模型组同样方法束缚20 min。治疗15天后,用Morris水迷宫检测小鼠的行为学变化,免疫组化观察小鼠额叶皮层小胶质细胞活化情况,Western Blot检测小鼠额叶皮层TNF-α的表达含量。结果:"通督启神"法不同电针均可显著改善APP/PS1小鼠的学习记忆能力(P<0.05),降低AD小鼠额叶皮层小胶质细胞标记物Iba-1的表达(P<0.05)及TNF-α蛋白的表达(P<0.05),且音乐电针组均优于脉冲电针组(P<0.05)。结论:"通督启神"法不同电针可有效改善APP/PS1小鼠学习记忆能力,抑制小胶质细胞活化、减少TNF-α的分泌,且音乐电针在改善额叶皮层炎性反应方面优于脉冲电针,对AD有更好的治疗效果。

This study aimed to investigate the effects of ＂Tongdu Qishen＂ of music electro-acupuncture （EA） combined with music therapy and EA combined with pulse therapy on the spatial learning and memory and the expression of microglia and TNF-α in the frontal cortex in APP/PS1 double transgenic mice, and to explore the their mechanisms underlying the treatment of EA for Alzheimer＇ s Disease （AD）. Twenty-four male APP/ PS1 transgenic mice were equally and randomly divided into the model group, the EA treatment group and the music EA group, while 8C57BL/6 mice were selected as the control group. In EA treatment group, GV20- Baihui and GV29-Yintang were stimulated by EA with the frequency of 2 Hz combined with music or pulse therapy, lasting for 20 rain every day. Then, swift pricking blood therapy was performed at GV26-Shuigou prior to EA treatment. Mice in the control group and the model group had been restrained for 20 min each day. After 15-day＇ s treatment, morris water maze test was adopted to observe the ethological changes of mice in each group. The expression of microglia in the frontal cortex was detected by immunohistochemistry technology, while western blot was applied to quantify TNF-α in frontal cortex. It was found that spatial learning and memory in mice of the music-EA group was improved compared with the EA group （P 〈 0.05）. Compared with the model group, the number of IBA-l-immunopositive cells was decreased in the EA group and the music-EA group （P 〈 0.05）. Western blot detection showed that TNF-α expression in frontal cortex of mice in the model group was higher than that in the control group （P 〈 0.05）. After the treatment, the TNF-α expression in both the EA group and the music-EA group was down-regulated. In conclusion, it was demonstrated that the treatments of pulse-EA and music-EA rested on ＂Tongdu Qishen＂ theory improved the spatial learning and memory of APP/PS1 double transgenic mice and inhibited the activated microglia and TNF-a expression in the frontal cortex. Music-EA therapy offered a better performance in allaying cerebral inflammation than pulse-EA treatment.