“醒脑开窍”针刺法调节ATP敏感性钾通道开放对抗脑缺血再灌注损伤的机制研究  

Mechanism of “Xingnao Kaiqiao” acupuncture for the opening of ATP sensitive potassium channel against cerebral ischemia reperfusion injury in rats

作　　者：韩林[1] 高旸[1] 王旭慧[1] 张亚男[2] 张向宇[1] HAN Lin;GAO Yang;WANG Xuhui;ZHANG Yanan;ZHANG Xiangyu(Acupuncture Department,the First Teaching Hospital of Tianjin University of TCM,Tianjin 300193,China;Acupuncture Research Institute,the First Teaching Hospital of Tianjin University of TCM)

机构地区：[1]天津中医药大学第一附属医院针灸部,天津300193 [2]天津中医药大学第一附属医院针灸研究所

出　　处：《中国针灸》2018年第12期1319-1324,共6页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金青年基金项目:81603688

摘　　要：目的:观察"醒脑开窍"针刺法调节ATP敏感性钾通道开放对抗脑缺血再灌注损伤的作用机制。方法:将84只大鼠随机分为假手术组、模型组、电针组、电针+三磷酸腺苷敏感性钾通道(K_(ATP))阻滞剂组(电针+K_(ATP)阻滞剂组),每组21只。电针+K_(ATP)阻滞剂组大鼠给予K_(ATP)通道阻断剂格列吡嗪(1μmol/5μL)10μL脑室内注入。模型组、电针组、电针+K_(ATP)阻滞剂组采用Zea Longa线栓法复制大脑中动脉缺血再灌注模型,假手术组仅分离颈总动脉及颈外动脉,不插入尼龙线栓。电针组、电针+K_(ATP)阻滞剂组给予针刺"内关""水沟""三阴交"治疗,留针20 min,留针期间将患侧"内关""三阴交"穴分别连接神经穴位刺激仪,施以疏密波,频率2 Hz/15 Hz,电流强度1 mA。首次针刺在动物造模成功90 min后进行,每天10:00、16:00各针刺1次,共3 d。假手术组、模型组大鼠同样抓取,但不予针刺。采用Zausinger六分法评价神经功能,2,3,5-氯化三苯基四氮唑(TTC)染色检测脑梗死体积,流式细胞技术检测海马神经细胞凋亡率,Western-blot技术检测B淋巴细胞瘤-2基因(Bcl-2)、B细胞淋巴瘤因子相关X蛋白(Bax)表达。结果:干预后,电针组神经功能评分明显高于模型组(P<0.01),脑梗死体积及海马神经细胞总凋亡率明显低于模型组(均P<0.05),电针组Bcl-2蛋白表达、Bcl-2/Bax比值较模型组均明显升高(P<0.05,P<0.01),Bax蛋白表达较模型组明显降低(P<0.01);与电针组比较,电针+K_(ATP)阻滞剂组大鼠神经功能评分明显降低(P<0.05),海马神经细胞总凋亡率明显升高(P<0.05),海马组织Bcl-2蛋白表达明显降低(P<0.05),Bax蛋白表达明显升高(P<0.05)。结论:"醒脑开窍"针刺法对脑缺血再灌注损伤大鼠有明显的脑保护作用,其机制可能与调节脑组织K_(ATP)通道开放、减轻神经细胞凋亡密切相关。Objective To observe the mechanism of “Xingnao Kaiqiao” acupuncture for the opening of ATP sensitive potassium channel (KATP) against cerebral ischemia reperfusion injury in rats. Methods Eighty-four rats were randomly divided into a sham-operation group, a model group, an electroacupuncture (EA) group, an EA+KATP blocker group, 21 rats in each group. 10 μL intracerebral injection with glipizide (1 μmol/5 μL) was used in the EA+KATP blocker group. The cerebral ischemia reperfusion model was established by Zea Longa's suture method in the model group, the EA group and the EA+KATP blocker group. Rats in the sham-operation group were received the same surgery but without nylon filament insertion. Acupuncture (20 min a time) was performed at “Neiguan” (PC 6), “Shuigou” (GV 26) and “Sanyinjiao” (SP 6) in the EA group and the EA+KATP blocker group at 10:00 and 16:00 for 3 days, firstly 90 min after model establishment. EA (2 Hz/15 Hz, 1 mA) was connected at the affected “Neiguan” (PC 6) and “Sanyinjiao” (SP 6). The same fixation was used in the sham-operation group and the model group, without EA. Neurological function was assessed by Zausinger's neurologic assessment scale. 2, 3, 5-triphenyltetrazolium chloride (TTC) staining was used to detect infarct volume. Neurocyte apoptosis in the hippocampus was detected by flow cytometry and the protein expressions of B lymphocytoma-2 gene (Bcl-2) and B cell lymphoma factor-associated X protein (Bax) were measured by Western-blot. Results In comparison with the model group, the neurological score of the EA group increased (P<0.01); the infarction volume and the hippocampal neuron’s total apoptosis rate of the EA group decreased (both P<0.05); the protein expression of Bcl-2 and Bcl-2/Bax of the EA group increased (P<0.05, P<0.01); and the protein expression of Bax of the EA group decreased (P<0.01). Compared with the EA group, the neurological score of the EA+KATP blocker group decreased (P<0.05); the total apoptosis rate of hippocampus neurons of

关 键 词：“醒脑开窍”针刺法 脑缺血再灌注 ATP敏感性钾通道 细胞凋亡 

分 类 号：R245.9[医药卫生—针灸推拿学]