Guo Fei, Zhang Su Zhao, Chen Shi Yu, Zhang Chuang, Zhang Xiao Qi, Gao Fei, Wu Ze Hui, Yu Wen Tao, Liang Yu Lei, Zhang Hui Zhen
College of Acupuncture and Moxibustion, Hebei University of Chinese Medicine, Shijia-zhuang 050200, China.
Department of Acupuncture and Moxibustion, Affiliated Hospital of Hebei University of Chinese Medicine, Shijiazhuang 050011.
Zhen Ci Yan Jiu. 2020 Jan 25;45(1):21-6. doi: 10.13702/j.1000-0607.1905126.
To observe the effect of electroacupuncture (EA) stimulation on the expression of c-Jun terminal kinase(JNK)signaling pathway-related proteins in the hippocampus of vascular dementia (VD) rats, so as to explore its mechanisms underlying improvement of VD.
Male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (＝10 rats per group). The VD model was prepared by repeated occlusion of the bilateral common carotid arteries for 10 min and reperfusion for 10 min (3 times in total). The rats in the EA group received EA (2 Hz, 2 mA) at "Dazhui"(GV14)，"Baihui"(GV20), and bilateral "Housanli"(ST36) ，"Geshu"(BL17) for 10 min, once daily for 14 days. The learning-memory abi-lity was detected by Morris water maze tests, the distribution of hippocampal neurons detected by Nissl staining, and the apoptosis of hippocampal neurons detected by using TdT-mediated dUTP nick-end labeling (TUNEL) method. The expressions of JNK, phosphorylated JNK (p-JNK), cysteine-containing aspartate-specific proteases-8 (Caspase-8) and Caspase-3 proteins were detected by Western blot.
After modeling and compared with the sham operation group, the escape latency was significantly prolonged (<0.01) and the number of safe-platform quadrant crossing obviously decreased (<0.01), suggesting a reduction of learning-memory ability. The number of hippocampal neurons was considerably reduced (<0.01), and that of hippocampal apoptotic neurons remarkably increased in the model group (<0.01). Whereas, the expression levels of hippocampal apoptosis-related proteins as JNK, p-JNK, Caspase-8 and Caspase-3, as well as the apoptotic index were significantly up-regulated (<0.01). Following EA intervention, the learning-memory ability was apparently improved (<0.01), and the number of hippocampal neurons was considerably increased (<0.01), the hippocampal apoptotic cell number, apoptosis index and the expression levels of JNK, p-JNK, Caspase-8 and Caspase-3 were significantly down-regulated (<0.01).
EA intervention can improve the learning-memory ability of VD rats, which may be associated with its effects in reducing hippocampal apoptosis by suppressing JNK signaling pathway.
观察电针刺激对血管性痴呆（VD）大鼠海马中c-Jun氨基末端激酶（JNK）信号通路相关蛋白表达的影响，以探讨其改善VD的机制。
将雄性Sprague-Dawley大鼠随机分为假手术组、模型组和电针组（每组n＝10只）。采用双侧颈总动脉反复阻断10分钟再灌注10分钟（共3次）的方法制备VD模型。电针组大鼠于“大椎”（GV14）、“百会”（GV20）及双侧“后三里”（ST36）、“膈俞”（BL17）接受电针刺激（2Hz，2mA），每次10分钟，每日1次，共14天。采用Morris水迷宫试验检测学习记忆能力，尼氏染色检测海马神经元分布，TdT介导的dUTP缺口末端标记（TUNEL）法检测海马神经元凋亡。采用蛋白质免疫印迹法检测JNK、磷酸化JNK（p-JNK）、含半胱氨酸的天冬氨酸特异性蛋白酶-8（Caspase-8）和Caspase-3蛋白的表达。
造模后与假手术组比较，逃避潜伏期显著延长（P<0.01），安全平台象限穿越次数明显减少（P<0.01），提示学习记忆能力下降。模型组海马神经元数量显著减少（P<0.01），海马凋亡神经元数量显著增加（P<0.01）。而海马凋亡相关蛋白JNK、p-JNK、Caspase-8和Caspase-3的表达水平以及凋亡指数均显著上调（P<0.01）。电针干预后，学习记忆能力明显改善（P<0.01），海马神经元数量显著增加（P<0.01），海马凋亡细胞数量、凋亡指数以及JNK、p-JNK、Caspase-8和Caspase-3的表达水平均显著下调（P<0.01）。
电针干预可改善VD大鼠的学习记忆能力，可能与其通过抑制JNK信号通路减少海马凋亡有关。
