摘要
目的:观察针刺对戊四唑(PTZ)诱发癫痫大鼠海马神经元损伤的保护作用及该保护作用与磷脂酰肌醇-3-激酶(PI 3K)/蛋白激酶B(Akt)信号转导通路的关系。方法:SD大鼠120只随机分为5组:正常组、PTZ组、LY 294002组、针刺+LY 294002组、针刺组,每组24只。针刺组、针刺+LY 294002组针刺"百会""大椎"30min,共治疗5次。正常组、PTZ组、针刺组先侧脑室注射二甲基亚砜5μL,LY 294002组、针刺+LY 294002组侧脑室注射LY 294002(5μL),30min后正常组腹腔注射生理盐水2mL,其余各组腹腔注射PTZ(50mg/kg),于注射后4h与24h取脑组织。分别用HE染色法于光镜下观察海马结构改变,用电镜观察海马超微结构。结果:癫痫发作后4h光镜、电镜均可见大鼠海马神经元损伤,24h后损伤进一步加重。LY 294002组大鼠海马神经元损伤明显加重。针刺+LY 294002组大鼠海马神经元损伤明显,与LY294002组比较未见明显好转。针刺组大鼠海马神经元损伤明显减轻。结论:针刺具有明显保护癫痫继发脑神经元损伤作用,其保护作用与细胞内PI 3K/Akt信号转导通路有关。
Objective 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. Methods 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 μL). Manual acupuncture stimulation was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) once daily for 5 days. Dimethyl Sulfoxide (DMSO, 5 μL, a control solvent) was given to rats of the normal, model and acupuncture groups, and LY294002 (5 μL, dissolved in DMSO) given to rats of the LY 294002 and acu- puncture+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 checking ultrastructural changes of the pyramidal cells under transmission electron microscope. Results In comparison with the normal control group, the numbers of pyramidal cellsof 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 microscopeshowed that the injury severity of pyramidal cells of h and even worse 24 h after seizure in the model group, in the acupuncture group. These results suggested an pathway by lateral ventricular injection of LY 294002 effect on pyramidal cells of hippocampal CA 1 and CA tracellular PI 3 K/Akt signaling pathway. ppocampal CA 1 and CA 3 regions was moderate 4 h after epileptic seizure LY 294002 group and acupuncture+ LY 294002 group, but relatively lighter elimination of the acupuncture effect after blocking the PI 3 K/Akt signaling 'n ep 3 reg epsy rats. Conclusion ons in epilepsy rats, which Acupuncture intervention has a protective s associated with the normal function of in-tracellular PI 3 K/Akt signaling pathway.
出处
《针刺研究》
CAS
CSCD
北大核心
2013年第1期20-25,共6页
Acupuncture Research
基金
安徽省自然科学基金(No.090413104)
国家自然科学基金(No.81072872)