Yang Fan, Ang Wen-Ping, Shen De-Kai, Liu Xiang-Guo, Yang Yong-Qing, Ma Yun
Research Institute of Acn-moxibustion and Meridian, Anhui College of Chinese Medicine, Hefei 230038, China.
Zhen Ci Yan Jiu. 2013 Feb;38(1):20-5.
To observe the protective effect of acupuncture stimulation on pyramidal cells in hippocampal CA 1 and CA 3 regions and to analyze the involvement of phosphatidy linositol-3-kinase (PI 3 K)/protein kinase B(PKB or Akt) signaling pathway in the acupuncture effect in epilepsy rats.
A total of 120 SD rats were randomly divided into normal control group, model group, LY 294002 (a specific antagonist for PI 3 K/Akt signaling) group, acupuncture+ LY 294002 group and acupuncture group (n = 24 in each group, 12 for H. E. staining, and 12 for electron microscope observation). Epilepsy model was established by intraperitoneal injection of pentylenetetrazol (PTZ, 5 microL). Manual acupuncture stimulation was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) once daily for 5 days. Dimethyl Sulfoxide (DMSO, 5 microL, a control solvent) was given to rats of the normal, model and acupuncture groups, and LY294002 (5 microL, dissolved in DMSO) given to rats of the LY 294002 and acupuncture+ LY 294002 groups by lateral ventricular injection. Four hours and 24 h after modeling, the hippocampus tissues were sampled for observing pathological changes of CA 1 and CA 3 regions after H. E. staining under light microscope and for checkin ultrastructural changes of the pyramidal cells under transmission electron microscope.
In comparison with the normal control group, the numbers of pyramidal cells of hippocampal CA 3 region in the model group were decreased significantly 4 h and 24 h after epileptic seizure (P < 0.01). While compared to the model group, the pyramidal cells of hippocampal CA 3 region in the acupuncture group were increased considerably in the number at both 4 h and 24 h after seizure (P < 0.01). No significant differences were found between the LY 294002 and model groups, and between the acupuncture+ LY 294002 and model groups in the numbers of pyramidal cells at 4 h and 24 h after seizure (P > 0.05). Findings of the light microscope and electron microscope showed that the injury severity of pyramidal cells of hippocampal CA 1 and CA 3 regions was moderate 4 h after epileptic seizure and even worse 24 h after seizure in the model group, LY 294002 group and acupuncture+ LY 294002 group, but relatively lighter in the acupuncture group. These results suggested an elimination of the acupuncture effect after blocking the PI 3 K/Akt signaling pathway by lateral ventricular injection of LY 294002 in epilepsy rats.
Acupuncture intervention has a protective effect on pyramidal cells of hippocampal CA 1 and CA 3 regions in epilepsy rats, which is associated with the normal function of intracellular PI 3 K/Akt signaling pathway.
观察针刺刺激对癫痫大鼠海马CA1和CA3区锥体细胞的保护作用，并分析磷脂酰肌醇-3-激酶（PI 3 K）/蛋白激酶B（PKB或Akt）信号通路在针刺抗癫痫效应中的作用机制。
将120只SD大鼠随机分为正常对照组、模型组、LY 294002（PI 3 K/Akt信号通路特异性拮抗剂）组、针刺+LY 294002组和针刺组（每组24只，其中12只用于苏木精-伊红染色，12只用于电镜观察）。采用腹腔注射戊四氮（PTZ，5 μL）制备癫痫模型。每天对“百会”（GV 20）和“大椎”（GV 14）进行一次手动针刺刺激，共5天。正常组、模型组和针刺组大鼠腹腔注射二甲基亚砜（DMSO，5 μL，作为对照溶剂），LY 294002组和针刺+LY 294002组大鼠侧脑室注射LY 294002（5 μL，溶于DMSO）。造模后4小时和24小时，取海马组织，光镜下观察苏木精-伊红染色后CA1和CA3区的病理变化，透射电镜下观察锥体细胞的超微结构变化。
与正常对照组相比，模型组癫痫发作后4小时和24小时海马CA3区锥体细胞数量显著减少（P<0.01）。与模型组相比，针刺组癫痫发作后4小时和24小时海马CA3区锥体细胞数量明显增加（P<0.01）。癫痫发作后4小时和24小时，LY 294002组与模型组、针刺+LY 294002组与模型组锥体细胞数量比较，差异无统计学意义（P>0.05）。光镜和电镜结果显示，模型组、LY 294002组和针刺+LY 294002组癫痫发作后4小时海马CA1和CA3区锥体细胞损伤程度为中度，24小时后损伤更严重，而针刺组损伤相对较轻。这些结果表明，侧脑室注射LY 294002阻断PI 3 K/Akt信号通路后，针刺抗癫痫效应消失。
针刺干预对癫痫大鼠海马CA1和CA3区锥体细胞具有保护作用，其机制可能与细胞内PI 3 K/Akt信号通路的正常功能有关。
