摘要
目的 评价电针预处理对小鼠脑缺血再灌注时海马神经元单磷酸腺苷激活的蛋白激酶（AMPK）活性的影响.方法 雄性C57BL6小鼠60只,7周龄,体重20～ 22 g,采用随机数字表法,将其分为5组（n=12）：正常对照组（C组）、假手术组（S组）、脑缺血再灌注组（I/R组）、电针刺激百会穴预处理组（EA＋I/R组）和电针刺激非穴位预处理组（NEA＋I/R组）.电针刺激百会穴预处理的实施：疏密波,频率2 Hz/15 Hz,强度1 mA,持续30 min,1次/d,连续5d,最后一次针刺结束后24 h制备脑缺血再灌注损伤模型.采用夹闭双侧颈总动脉15 min后恢复脑血流的方法建立脑缺血再灌注损伤模型.术后3d时进行神经行为学缺陷（NDS）评分,随后处死,取海马组织,光镜下观察病理学结果,采用TUNEL法检测凋亡神经元,采用Western blot法检测磷酸化AMPKα（pAMPKα）和caspase-3的表达.结果 与C组比较,I/R组和EA＋ I/R组NDS评分和海马CA1区凋亡细胞计数升高,海马pAMPKα和caspase-3上调（P＜0.05）,S组上述指标差异无统计学意义（P＞0.05）;与I/R组比较,EA＋I/R组NDS评分和和海马CA1区凋亡细胞计数降低,海马pAMPKα表达上调,海马caspase-3表达下调（P＜0.05）,NEA＋I/R组述指标差异无统计学意义（P＞0.05）.EA＋I/R组海马CA1区病理学损伤明显轻于I/R组.结论 电针预处理减轻小鼠脑缺血再灌注时海马神经元凋亡的机制与促进AMPK激活有关.
Objective To evaluate the effect of electroacupuncture （EA） preconditioning on activity of AMP-activated protein kinase （AMPK） in hippocampal neurons during cerebral ischemiareperfusion （I/R） in mice.Methods A total of 60 male C57BL6 mice,aged 7 weeks,weighing 20-22 g,were randomly divided into 5 groups （n=12 each） using a random number table：control group （C group）,sham operation group （S group）,I/R group,acupuncture at acupoint Baihui preconditioning group （EA ＋ I/R group）,and acupuncture at non-acupoint preconditioning group （NEA ＋ I/R group）.Baihui acupoints were stimulated with electric stimulator （frequency 2 Hz/15 Hz,intensity 1 mA） for 30 min once a day for 5 consecutive days.At 24 h after the last stimulation,the model of cerebral I/R injury was established.Bilateral common carotid arteries were occluded by clipping for 15 min followed by reperfusion.Neurological deficit score （NDS） was assessed at 3 days after operation.Then the mice were sacrificed and the brains were immediately harvested for microscopic examination and for determination of apoptosis in hippocampal neurons （using TUNEL） and expression of phosphor-AMPKα （pAMPKα） and caspase-3 （by Western blot）.Results Compared with group C,NDS and the number of apoptotic neurons in hippocampal CA1 region were significantly increased,and the expression of pAMPKα and caspase-3 was up-regulated in I/R and EA＋I/R groups,and no significant change was found in the parameters mentioned above in group S.Compared with group I/R,NDS and the number of apoptotic neurons in hippocampal CA1 region were significantly increased,the expression of pAMPKα was up-regulated,and the expression of caspase-3 was down-regulated in group EA ＋I/R,and no significant change was found in the parameters mentioned above in group NEA＋I/R.The pathological changes in hippocampal CA1 region were significantly attenuated in group EA ＋ I/R as compared with group I/R.Conclusion The mechanism by which EA preconditioning mi
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2015年第1期44-47,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金（81273821）
关键词
电刺激疗法
脑
再灌注损伤
环AMP依赖性蛋白激酶类
Electric stimulation therapy
Brain
Reperfusion injury
Cyclic AMP-dependent protein kinases
