“醒脑开窍”针刺法对脑缺血再灌注大鼠海马神经元KATP通道细胞电生理的调控研究

Regulation of “Xingnao Kaiqiao” acupuncture therapy on cell-electrophysiology of KATP channel in hippocampal neurons of rats with cerebral ischemia reperfusion injury

关键词:  “醒脑开窍”针刺法  脑缺血再灌注  ATP敏感性钾通道  细胞电生理  膜片钳

Key words:  “Xingnao Kaiqiao” acupuncture therapy  cerebral ischemia reperfusion  ATP sensitive potassium channel  cell-electrophysiology  patch-clamp

摘要:

[目的]观察"醒脑开窍"针刺法对脑缺血再灌注大鼠海马椎体神经元三磷酸腺苷敏感性钾（K_ATP）通道电生理特性的影响。[方法]将32只大鼠随机分为空白组、假手术组、模型组、电针组，每组8只。模型组、电针组大鼠按照Zea longa线栓法复制大脑中动脉缺血再灌注模型，假手术组只分离颈总动脉和颈外动脉，不插入尼龙线栓。电针组给予针刺内关、水沟、三阴交治疗，每日2次，连续治疗3 d。采用Zausinger六分法评价神经功能，神经元急性分离技术获得海马椎体神经元，采用单细胞膜片钳技术记录各组大鼠海马椎体神经元K_ATP通道电流曲线，各组随机选择封接成功的8个膜片进行膜片钳数据分析。[结果]模型组大鼠神经功能评分明显低于空白组、假手术组（均P<0.01），海马锥体神经元K_ATP通道开放时间、开放概率均明显高于空白组、假手术组（均P<0.01），电流幅度与空白组、假手术组比较均无统计学差异（均P>0.05）；电针组大鼠神经功能评分明显高于模型组（P<0.01），海马锥体神经元K_ATP通道开放时间、开放概率均明显低于模型组（均P<0.01），电流幅度与模型组比较无统计学差异（P>0.05）。[结论]"醒脑开窍"针刺法可以影响脑缺血再灌注大鼠海马椎体神经元K_ATP通道的电生理特性，其对抗脑缺血再灌注损伤的作用机制可能与调节K_ATP通道开放活动相关。

Abstract:

[Objective] To observe the effect of "Xingnao Kaiqiao" acupuncture therapy on electrophysiological characteristics of K_ATP channel in hippocampal vertebral neurons of rats with cerebral ischemia reperfusion injury.[Method] The 32 Wistar rats were randomly divided into four groups,the normal group,the sham-operation group,the I/R group and the electroacupuncture group,8 rats in each group. The cerebral ischemia reperfusion model was established by Longa's suture method in model group and the electroacupuncture (EA) group. Rats in sham-operated group received the same surgery,but without nylon filament insertion. In electroacupuncture group,acupuncture was performed with Xingnao Kaiqiao method at main acupoints of Neiguan (PC6),Shaigou (GV26),Sanyinjiao (SP6) and the electric acupuncture apparatus was connected to Neiguan (PC6) and Sanyinjiao (SP6) on the affected side. Then acupunctures were followed twice daily for 3 days. Neurological deficits were assessed by ethologic scoring according to Zausinger's neurologic assessment scales,hippocampal vertebral neurons were obtained by neuronal acute dissociation technique and the single cell patch clamp technique was used to record the K_ATP current curve of hippocampal vertebral neurons in each group. Then,8 successfully sealed patches were selected randomly in each group for patch-clamp data processing.[Results] Neurological deficits score in model group was significantly lower than that in blank group and sham operation group(P<0.01),the open time and open probability of K_ATP channel in hippocampal pyramidal neurons of model group were significantly higher than those of blank group and sham operation group(P<0.01)and the amplitude of the current in the model group was not significantly different from that in the blank group and the sham operation group(P>0.05). The neurological deficits score of electroacupuncture group was significantly higher than that of model group(P<0.01),the K_ATP channel open time and open probability of hippocampal pyramidal neurons in electroacupuncture group were significantly lower than those in model group(P<0.01)and there was no significant difference in the current amplitude between the EA group and the model group (P>0.05).[Conclusions] "Xingnao Kaiqiao" acupuncture therapy can affect the electrophysiological characteristics of K_ATP channels in hippocampal neurons of rats with cerebral ischemia-reperfusion injury,and the underlying mechanism of "Xingnao Kaiqiao" acupuncture therapy against cerebral ischemia reperfusion injury may be related to the regulation of K_ATP channel opening activities.