Huang Rui, Gong Xin, Ni Jin-Zhong, Jia Yuan-Wei, Zhao Jian
Department of Anatomy, Wannan Medical College, Wuhu 241002, Anhui Province, China.
Yijishan Hospital of Wannan Medical College.
Zhongguo Zhen Jiu. 2019 Apr 12;39(4):397-402. doi: 10.13703/j.0255-2930.2019.04.014.
To explore the mechanism of acupuncture plus medication on treatment of Alzheimer's disease (AD).
Sixty adult SD rats were randomly divided into a normal group, a sham operation group, a model group, an electroacupuncture (EA) group, a gastrodin group and an EA+gastrodin group, 10 rats in each one. The rat model of AD was established by intraperitoneal injection of D-galactose and bilateral hippocampal injection of Aβ1-40. Two weeks after modeling, the rats in the EA group and EA+gastrodin group were treated with EA at "Baihui" (GV 20) "Dazhui" (GV 14) and bilateral "Zusanli" (ST 36), 30 min per treatment, once a day for consecutive 4 weeks. The rats in the gastrodin group and EA+gastrodin group were treated with intraperitoneal injection of gastrodin, once a day for consecutive 4 weeks. The rats in the normal group, model group and sham operation group were not treated. The morphology of hippocampal neurons was observed by using HE staining. The expression of Bcl-2 and Bax in the hippocampal CA1 area was detected by using immunohistochemical method. The expression of Bcl-2 and Bax protein in hippocampus was detected by using Western blot.
The HE staining results showed the arrangement of neurons in the hippocampal CA1 area was regular in the normal group and the sham operation group, and the cytoplasm and nucleus were clearly visible. The neurons in the model group were severely damaged; the cell arrangement was not close, and the cell morphology was incomplete. Compared with the model group, the cell morphology of each intervention group was significantly improved. The immunohistochemistry results showed that, compared with the normal group and the sham operation group, the expression of Bcl-2 in the hippocampal CA1 region in the model group was decreased (<0.05), but the expression of Bax was enhanced (<0.05); compared with the model group, the expression of Bcl-2 was increased (all <0.05) and the expression of Bax was decreased (all <0.05) in all intervention group; compared with the EA group or the gastrodin group, the expression of Bcl-2 was enhanced (<0.05) and the expression of Bax was decreased (<0.05) in the EA+gastrodin group. The result of Western blot method was consistent with that of immunohistochemistry method.
EA and gastrodin could significantly inhibit the expression of Bax and up-regulate the expression of Bcl-2, and the combination of EA and gastrodin has the most significant effect. This indicates that EA combined with gastrodin has synergistic effect on inhibiting the apoptosis of neurons in hippocampus in AD rats, which may be one of the mechanisms of EA plus medication on AD lesions.
探讨针刺联合药物治疗阿尔茨海默病（AD）的机制。
将60只成年SD大鼠随机分为正常组、假手术组、模型组、电针组、天麻素组和电针+天麻素组，每组10只。采用腹腔注射D-半乳糖和双侧海马注射Aβ1-40建立AD大鼠模型。造模后2周，电针组和电针+天麻素组大鼠于“百会”（GV 20）、“大椎”（GV 14）及双侧“足三里”（ST 36）进行电针治疗，每次治疗30分钟，每天1次，连续4周。天麻素组和电针+天麻素组大鼠腹腔注射天麻素，每天1次，连续4周。正常组、模型组和假手术组大鼠不做处理。采用HE染色观察海马神经元形态。采用免疫组化法检测海马CA1区Bcl-2和Bax的表达。采用蛋白质免疫印迹法检测海马中Bcl-2和Bax蛋白的表达。
HE染色结果显示，正常组和假手术组海马CA1区神经元排列规则，细胞质和细胞核清晰可见。模型组神经元严重受损；细胞排列不紧密，细胞形态不完整。与模型组相比，各干预组细胞形态均有明显改善。免疫组化结果显示，与正常组和假手术组相比，模型组海马CA1区Bcl-2表达降低（<0.05），但Bax表达增强（<0.05）；与模型组相比，各干预组Bcl-2表达均升高（均<0.05），Bax表达均降低（均<0.05）；与电针组或天麻素组相比，电针+天麻素组Bcl-2表达增强（<0.05），Bax表达降低（<0.05）。蛋白质免疫印迹法结果与免疫组化法结果一致。
电针和天麻素可显著抑制Bax表达，上调Bcl-2表达，且电针与天麻素联合应用效果最为显著。这表明电针联合天麻素对抑制AD大鼠海马神经元凋亡具有协同作用，可能是针刺联合药物治疗AD损伤的机制之一。
