基于钠钾ATP酶-兴奋性氨基酸转运体-谷氨酸通路的针刺治疗脑卒中后痉挛中枢机制研究

[Study on the central mechanism of acupuncture for post-stroke spasticity based on the Na/K-ATPase-EAATs-Glu pathway].

作者信息

Qian Xu, Ma Liang-Xiao, Mu Jie-Dan, Zhang Zhou, Sun Tian-Yi, Yu Wen-Yan, Tian Yuan, Zhang Yi-Dan

机构信息

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China.

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China; The Key Unit of State Administration of Traditional Chines Medicine, Evaluation of Characteristic Acupuncture Therapy, Beijing 100029.

出版信息

Zhen Ci Yan Jiu. 2022 Apr 25;47(4):283-9. doi: 10.13702/j.1000-0607.20210922.

DOI:10.13702/j.1000-0607.20210922

PMID:35486007

Abstract

OBJECTIVE

To observe the effect of acupuncture at "Yanglingquan" (GB34) and "Baihui" (GV20) on Na/K-ATPase, excitatory amino acid transporters (EAATs) and glutamate (Glu) in hippocampus of post-stroke spasticity rats, so as to explore the central mechanism in anti-spasticity.

METHODS

In a total of 48 healthy SD rats, 12 rats were randomly selected to be included into sham operation group, and the remaining rats were used to make a middle cerebral artery occlusion (MCAO) model using a suture method. On the 3rd day after modeling, MCAO limb spasticity rats were screened by neurological deficit symptoms and muscle tension scores, and randomly divided into the model, GB34 (Hui-puncture at GB34) and GB34+GV20 (Hui-puncture at GB34 and horizontal insertion at GV20) groups (=12 rats in each group), and the treatment was lasted for 7 conse-cutive days. The neurological symptoms and muscle tension score were observed with the Zea Longa score and modified Ashworth scale (MAS). The levels of Glu, EAAT1 (GLAST) and EAAT2 (GLT-1) in the ischemic area of cerebral hippocampus were detected by ELISA, the expression of Na/K-ATPase α1 (ATP1α1) was detected by Western blot, the expression of ATP1α1 mRNA was detected by real-time PCR, and the expression of GLAST, GLT-1 and ATP1α1 was detected by immunofluorescence.

RESULTS

After modeling, Zea Longa score and MAS score were increased (<0.01), the level of Glu in the ischemic area of cerebral hippocampus was increased (<0.01), while the expression levels of GLAST, GLT-1, ATP1α1 protein and mRNA were all decreased (<0.01) in the model group relevant to the sham operation group. After 7 days' treatment, all the increased and decreased levels of the indexes mentioned above were reversed in the two acupuncture groups relevant to the model group (<0.01, <0.05), and the effects of acupuncture at GB34+GV20 were obviously superior to that of acupuncture at GB34 (<0.05, <0.01).

CONCLUSION

Acupuncture can alleviate post-stroke spasticity effectively, which may be related to its effect in up-regulating the expressions of Na/K-ATPase and EAATs in hippocampus. The anti-spastic effect of acupuncture at GB34+GV20 is superior to GB34 alone.

摘要

目的

观察针刺“阳陵泉”（GB34）与“百会”（GV20）对脑卒中后痉挛大鼠海马组织中钠钾-ATP酶、兴奋性氨基酸转运体（EAATs）及谷氨酸（Glu）的影响，以探讨针刺抗痉挛的中枢机制。

方法

选取48只健康SD大鼠，随机选取12只纳入假手术组，其余大鼠采用线栓法制备大脑中动脉闭塞（MCAO）模型。造模后第3天，通过神经功能缺损症状及肌张力评分筛选出MCAO肢体痉挛大鼠，随机分为模型组、GB34组（针刺GB34）和GB34+GV20组（针刺GB34并GV20平刺），每组12只，连续治疗7天。采用Zea Longa评分和改良Ashworth量表（MAS）观察神经症状和肌张力评分。采用ELISA法检测脑海马缺血区Glu、EAAT1（GLAST）和EAAT2（GLT-1）水平，采用Western blot法检测钠钾-ATP酶α1（ATP1α1）表达，采用实时荧光定量PCR法检测ATP1α1 mRNA表达，采用免疫荧光法检测GLAST、GLT-1和ATP1α1表达。

结果

造模后，模型组Zea Longa评分和MAS评分升高（P<0.01），脑海马缺血区Glu水平升高（P<0.01），而与假手术组相比，模型组GLAST、GLT-1、ATP1α1蛋白及mRNA表达水平均降低（P<0.01）。治疗7天后，与模型组相比，两个针刺组上述指标升高及降低水平均得到逆转（P<0.01，P<0.05），且针刺GB34+GV20组效果明显优于针刺GB34组（P<0.05，P<0.01）。