基于Na~+/K~+-ATP酶-谷氨酸转运体-谷氨酸途径探讨针刺治疗卒中后痉挛的中枢机制

钱旭; 马良宵; 母杰丹; 张洲; 孙天祎; 于文颜; 田园; 张一丹

doi:10.13702/j.1000-0607.20210922

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基于Na~+/K~+-ATP酶-谷氨酸转运体-谷氨酸途径探讨针刺治疗卒中后痉挛的中枢机制

机制探讨 | 更新时间：2023-08-11

- 基于Na~+/K~+-ATP酶-谷氨酸转运体-谷氨酸途径探讨针刺治疗卒中后痉挛的中枢机制
- Study on the central mechanism of acupuncture for post-stroke spasticity based on the Na~+/K~+-ATPase-EAATs-Glu pathway
- 针刺研究 2022年47卷第4期页码：283-289
- 作者机构：
  
  1. 北京中医药大学针灸推拿学院
  2. 国家中医药管理局针灸特色疗法评价重点研究室
- 作者简介：
- 基金信息：
  
  中央高校基本科研业务费专项资金项目(No.2020-JYB-ZDGG-062);国家自然科学基金项目(No.81774417)
- DOI：10.13702/j.1000-0607.20210922
  中图分类号：
- 纸质出版日期：2022
- 稿件说明：
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钱旭, 马良宵, 母杰丹, 等. 基于Na~+/K~+-ATP酶-谷氨酸转运体-谷氨酸途径探讨针刺治疗卒中后痉挛的中枢机制[J]. 针刺研究, 2022,47(4):283-289.

QIAN Xu, MA Liang-xiao, MU Jie-dan, et al. Study on the central mechanism of acupuncture for post-stroke spasticity based on the Na~+/K~+-ATPase-EAATs-Glu pathway[J]. Acupuncture research, 2022, 47(4): 283-289.
钱旭, 马良宵, 母杰丹, 等. 基于Na~+/K~+-ATP酶-谷氨酸转运体-谷氨酸途径探讨针刺治疗卒中后痉挛的中枢机制[J]. 针刺研究, 2022,47(4):283-289. DOI： 10.13702/j.1000-0607.20210922.

QIAN Xu, MA Liang-xiao, MU Jie-dan, et al. Study on the central mechanism of acupuncture for post-stroke spasticity based on the Na~+/K~+-ATPase-EAATs-Glu pathway[J]. Acupuncture research, 2022, 47(4): 283-289. DOI： 10.13702/j.1000-0607.20210922.

摘要

目的:观察针刺对卒中后痉挛大鼠海马区Na

/K+/K

-ATP酶、谷氨酸转运体(EAATs)和谷氨酸(Glu)的影响，探讨针刺抗痉挛的中枢机制。方法:SD大鼠随机分为假手术组、模型组、“阳陵泉”组和“阳陵泉”+“百会”组，每组12只。改良线栓法制作大鼠局灶性脑缺血模型。“阳陵泉”组恢刺患肢“阳陵泉”

“阳陵泉”+“百会”组恢刺患肢“阳陵泉”再平刺“百会”

均留针30 min

每日治疗1次，连续7 d。记录各组大鼠神经功能损伤(Zea Longa)评分和痉挛状态(MAS)评分，采用ELISA法检测缺血侧海马区Glu、EAAT1(GLAST)和EAAT2(GLT-1)含量，Western blot法检测缺血侧海马区Na+-ATP酶、谷氨酸转运体(EAATs)和谷氨酸(Glu)的影响，探讨针刺抗痉挛的中枢机制。方法:SD大鼠随机分为假手术组、模型组、“阳陵泉”组和“阳陵泉”+“百会”组，每组12只。改良线栓法制作大鼠局灶性脑缺血模型。“阳陵泉”组恢刺患肢“阳陵泉”

“阳陵泉”+“百会”组恢刺患肢“阳陵泉”再平刺“百会”

均留针30 min

每日治疗1次，连续7 d。记录各组大鼠神经功能损伤(Zea Longa)评分和痉挛状态(MAS)评分，采用ELISA法检测缺血侧海马区Glu、EAAT1(GLAST)和EAAT2(GLT-1)含量，Western blot法检测缺血侧海马区Na

/K+/K

-ATP酶α1(ATP1α1)的蛋白表达，实时荧光定量PCR法检测缺血侧海马区ATP1α1 mRNA表达，免疫荧光法检测缺血侧海马区GLAST、GLT-1、ATP1α1的表达。结果:与假手术组比较，模型组Zea Longa与MAS评分均升高(P

0.01);与模型组比较，两针刺组Zea Longa与MAS评分均降低(P

0.01);且“阳陵泉”+“百会”组低于“阳陵泉”组(P

0.05)。与假手术组比较，模型组缺血侧海马区Glu含量升高(P

0.01)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均降低(P

0.01);与模型组比较，两针刺组缺血侧海马区Glu含量降低(P

0.05

0.01)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均升高(P

0.05

0.01);与“阳陵泉”组比较，“阳陵泉”+“百会”组缺血侧海马区Glu含量降低(P

0.05)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均升高(P

0.05

0.01)。结论:针刺抗卒中后痉挛的效应可能与上调海马Na+-ATP酶α1(ATP1α1)的蛋白表达，实时荧光定量PCR法检测缺血侧海马区ATP1α1 mRNA表达，免疫荧光法检测缺血侧海马区GLAST、GLT-1、ATP1α1的表达。结果:与假手术组比较，模型组Zea Longa与MAS评分均升高(P

0.01);与模型组比较，两针刺组Zea Longa与MAS评分均降低(P

0.01);且“阳陵泉”+“百会”组低于“阳陵泉”组(P

0.05)。与假手术组比较，模型组缺血侧海马区Glu含量升高(P

0.01)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均降低(P

0.01);与模型组比较，两针刺组缺血侧海马区Glu含量降低(P

0.05

0.01)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均升高(P

0.05

0.01);与“阳陵泉”组比较，“阳陵泉”+“百会”组缺血侧海马区Glu含量降低(P

0.05)

GLAST、GLT-1含量及蛋白表达，ATP1α1蛋白和mRNA表达均升高(P

0.05

0.01)。结论:针刺抗卒中后痉挛的效应可能与上调海马Na

/K+/K

-ATP酶和EAATs的表达有关，恢刺“阳陵泉”加平刺“百会”的抗痉挛效应优于单纯恢刺“阳陵泉”穴。Objective To observe the effect of acupuncture at “Yanglingquan”(GB34) and “Baihui”(GV20) on Na+-ATP酶和EAATs的表达有关，恢刺“阳陵泉”加平刺“百会”的抗痉挛效应优于单纯恢刺“阳陵泉”穴。

Abstract

Objective To observe the effect of acupuncture at “Yanglingquan”(GB34) and “Baihui”(GV20) on Na

/K+/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 3+-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 3

(rd)

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(n=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(rd) 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(n=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+/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(P

0.01)

the level of Glu in the ischemic area of cerebral hippocampus was increased(P

0.01)

while the expression levels of GLAST

GLT-1

ATP1α1 protein and mRNA were all decreased(P

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(P

0.01

0.05)

and the effects of acupuncture at GB34+GV20 were obviously superior to that of acupuncture at GB34(P

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+-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(P

0.01)

the level of Glu in the ischemic area of cerebral hippocampus was increased(P

0.01)

while the expression levels of GLAST

GLT-1

ATP1α1 protein and mRNA were all decreased(P

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(P

0.01

0.05)

and the effects of acupuncture at GB34+GV20 were obviously superior to that of acupuncture at GB34(P

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+/K

-ATPase and EAATs in hippocampus. The anti-spastic effect of acupuncture at GB34+GV20 is superior to GB34 alone.

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