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["针刺通过开放ATP敏感性钾通道抗大鼠脑缺血再灌注损伤的机制研究"]

[Mechanism of "" acupuncture for the opening of ATP sensitive potassium channel against cerebral ischemia reperfusion injury in rats].

作者信息

Han Lin, Gao Yang, Wang Xuhui, Zhang Yanan, Zhang Xiangyu

机构信息

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.

出版信息

Zhongguo Zhen Jiu. 2018 Dec 12;38(12):1319-24. doi: 10.13703/j.0255-2930.2018.12.017.

Abstract

OBJECTIVE

To observe the mechanism of "" acupuncture for the opening of ATP sensitive potassium channel (K) 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+K blocker group, 21 rats in each group. 10 μL intracerebral injection with glipizide (1 μmol/5 μL) was used in the EA+K 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+K 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+K 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 (<0.01); the infarction volume and the hippocampal neuron's total apoptosis rate of the EA group decreased (both <0.05); the protein expression of Bcl-2 and Bcl-2/Bax of the EA group increased (<0.05, <0.01); and the protein expression of Bax of the EA group decreased (<0.01). Compared with the EA group, the neurological score of the EA+K blocker group decreased (<0.05); the total apoptosis rate of hippocampus neurons of the EA+K blocker group increased (<0.05); the expression of Bcl-2 protein of the EA+K blocker group reduced (<0.05); the expression of Bax protein of the EA+K blocker group increased (<0.05).

CONCLUSION

"" acupuncture has brain protective effect on rats with focal cerebral ischemia reperfusion injury. The mechanism may be related to regulating the opening of K channels and decreasing the apoptosis of neurons.

摘要

目的

观察“针刺开放ATP敏感性钾通道(K)”抗大鼠脑缺血再灌注损伤的机制。

方法

84只大鼠随机分为假手术组、模型组、电针组、电针+K通道阻滞剂组,每组21只。电针+K通道阻滞剂组脑室内注射10 μL格列吡嗪(1 μmol/5 μL)。模型组、电针组和电针+K通道阻滞剂组采用Zea Longa线栓法建立脑缺血再灌注模型。假手术组大鼠接受相同手术,但不插入尼龙丝。电针组和电针+K通道阻滞剂组于造模后90 min开始,在“内关”(PC 6)、“水沟”(GV 26)和“三阴交”(SP 6)穴针刺(每次20 min),每天10:00和16:00各针刺1次,共3天。在患侧“内关”(PC 6)和“三阴交”(SP 6)连接电针(2 Hz/15 Hz,1 mA)。假手术组和模型组采用相同固定方式,不进行电针治疗。采用Zausinger神经功能评分量表评估神经功能。用2,3,5-氯化三苯基四氮唑(TTC)染色检测梗死体积。采用流式细胞术检测海马神经细胞凋亡,并用蛋白质免疫印迹法检测B淋巴细胞瘤-2基因(Bcl-2)和B细胞淋巴瘤因子相关X蛋白(Bax)的蛋白表达。

结果

与模型组比较,电针组神经功能评分升高(<0.01);梗死体积及海马神经元总凋亡率降低(均<0.05);Bcl-2蛋白表达及Bcl-2/Bax升高(<0.05,<0.01);Bax蛋白表达降低(<0.01)。与电针组比较,电针+K通道阻滞剂组神经功能评分降低(<0.05);海马神经元总凋亡率升高(<0.05);Bcl-2蛋白表达降低(<0.05);Bax蛋白表达升高(<0.05)。

结论

“针刺”对大鼠局灶性脑缺血再灌注损伤具有脑保护作用。其机制可能与调节K通道开放及减少神经元凋亡有关。

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