Zhang Song-Jiang, Sun Ning-Ning, Gao Jian-Feng
School of Basic Medicine, Henan University of CM, Zhengzhou 450046, China.
Zhongguo Zhen Jiu. 2021 Mar 12;41(3):295-302. doi: 10.13703/j.0255-2930.20200203-k0002.
To screen protein target in prevention and treatment with electroacupuncture (EA) for Alzheimer's disease (AD) and explore the potential mechanism of EA in prevention of AD.
A total of 40 APP/PS1 transgenic young male mice, 1.5-month old, were randomized into an EA group and a model group, 20 mice in each one, and 20 C57BL/6J mice were chosen as the normal control group. After adaptive housing for 1 week, the mice in the EA group were stimulated with EA at "Baihui" (GV 20), "Fengfu" (GV 16) and "Shenshu" (BL 23), with intermittent wave, 10 Hz in frequency and 2 mA in electric intensity. EA was given once daily, 20 min each time. There was 1 day at interval after EA for 6 days each week. Totally, the intervention lasted for 16 weeks. On day 3 after the end of EA intervention, Morris water maze test was adopted to detect learning and memory abilities of mice in each group. After water maze test, the label-free method was used to measure the difference expressions in cerebral cortex and hippocampus. Using Western blot method, the expressions of guanylate binding protein beta 5 (GNB 5) and histone-H 3 in cerebral cortex and hippocampus were verified. Using immunohistochemical method, the expressions of amyloid beta protein (Aβ) in cerebral cortex and hippocampus were detected.
Compared with the normal control group, the escape latency (on day 2, 3 and 4) was prolonged, the frequency of crossing platform and the duration of platform stay were decreased in the mice of the model group (<0.05). Compared with the model group, the escape latency (on day 3 and 4) was shortened, the frequency of crossing platform and the duration of platform stay were increased in the mice of the EA group (<0.05). By the comparison among the three groups, the high mobility group nucleosome-binding domain-containing protein 5, band 3 anion transport protein, histone-H 3, epoxide hydrolase 4 (fragment), neurolysin (mitochondria), phosphoglycerate mutase 2, GNB5 and Aβ were the differential proteins with the larger fold-change difference in expression. Compared with the normal control group, the expression of histone-H 3 in cerebral cortex and hippocampus was reduced (<0.001) and the expressions of GNB 5 and Aβ were increased (<0.001, <0.01) in the mice of the model group. Compared with the model group, the expression of histone-H 3 in cerebral cortex and hippocampus was increased (<0.001) and the expressions of GNB 5 and Aβwere reduced (<0.001, <0.05) in the mice of the EA group.
The intervention with EA effectively prevents from the decline of learning and memory ability and the formation of Aβ senile plaques in cerebral cortex and hippocampus in young mouse models of AD after growing up. Besides, EA plays a regulatory function for protein expression differences induced by AD model.
筛选电针（EA）防治阿尔茨海默病（AD）的蛋白质靶点，探讨EA预防AD的潜在机制。
选取40只1.5月龄的APP/PS1转基因雄性幼鼠，随机分为电针组和模型组，每组20只，另选20只C57BL/6J小鼠作为正常对照组。适应性饲养1周后，电针组小鼠于“百会”（GV 20）、“风府”（GV 16）和“肾俞”（BL 23）进行电针刺激，采用疏密波，频率10Hz，强度2mA。每天电针1次，每次20分钟。每周电针6天，间隔1天。干预共持续16周。电针干预结束后第3天，采用Morris水迷宫试验检测各组小鼠的学习记忆能力。水迷宫试验后，采用无标记法检测大脑皮质和海马区的差异表达。采用Western blot法验证大脑皮质和海马区鸟苷酸结合蛋白β5（GNB 5）和组蛋白-H 3的表达。采用免疫组化法检测大脑皮质和海马区淀粉样β蛋白（Aβ）的表达。
与正常对照组相比，模型组小鼠逃避潜伏期（第2、3和4天）延长，穿越平台次数和在平台停留时间减少（<0.05）。与模型组相比，电针组小鼠逃避潜伏期（第3和4天）缩短，穿越平台次数和在平台停留时间增加（<0.05）。三组比较，高迁移率族核小体结合域蛋白5、带3阴离子转运蛋白、组蛋白-H 3、环氧水解酶4（片段）、神经溶素（线粒体）、磷酸甘油酸变位酶2、GNB5和Aβ是表达倍数变化差异较大的差异蛋白。与正常对照组相比，模型组小鼠大脑皮质和海马区组蛋白-H 3表达降低（<0.001），GNB 5和Aβ表达增加（<0.001，<0.01）。与模型组相比，电针组小鼠大脑皮质和海马区组蛋白-H 3表达增加（<0.001），GNB 5和Aβ表达降低（<0.001，<0.05）。
电针干预可有效预防成年后AD幼鼠模型学习记忆能力下降及大脑皮质和海马区Aβ老年斑形成。此外，电针对AD模型诱导的蛋白质表达差异具有调节作用。
