[电针“水沟”（GV26）和“百会”（GV20）对脑缺血再灌注损伤大鼠海马神经元自噬的影响]

[Effect of electroacupuncture at "Shuigou" (GV26) and "Baihui" (GV20) on autophagy of hippo-campal neurons in rats with cerebral ischemia-reperfusion injury].

作者信息

Liu Dan-Ni, Zhou Jun, Huang Xia-Rong, Sun Guang-Hua, Luo Fu, Tan Jin-Qu, Liao Yuan, Peng Ting, Feng Wei-Bin

机构信息

Rehabilitation Medicine Center, The First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province, China; Department of Rehabilitation, The First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province, China; Rehabilitation Laboratory, The First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province, China.

出版信息

Zhen Ci Yan Jiu. 2022 Jun 25;47(6):491-6. doi: 10.13702/j.1000-0607.20210561.

DOI:10.13702/j.1000-0607.20210561

PMID:35764515

Abstract

OBJECTIVE

To explore the effect of electroacupuncture (EA) at "Shuigou"(GV6) and "Baihui"(GV20) on autophagy of hippocampal neurons in cerebral ischemia-reperfusion (I/R) injury rats.

METHODS

Forty-eight healthy male SD rats were randomly divided into sham operation, model and EA groups, with 16 rats in each group. The rat model of cerebral I/R injury was established by occlusion of the middle cerebral artery (MCAO). Rats of the EA group received EA at GV26 and GV20 for 20 min, once daily for 5 days. The neurological function of rats in each group was evaluated by Longa neurological function score. The cerebral infarction volume was measured by TTC staining. The levels of IL-6, IL-18 and TNF-α in cerebrospinal fluid were detected by ELISA. Real-time PCR and Western blot were respectively used to detect the expressions of autophagy-related proteins AMPK, Beclin-1, VPS34 and LC3B.

RESULTS

Compared with the sham operation group, neurological function scores of rats in the model group were significantly increased (<0.01); the volume of cerebral infarction was significantly increased (<0.01); the contents of IL-6, IL-18 and TNF-α in cerebrospinal fluid were increased (<0.01, <0.05); the mRNA expression levels of AMPK, Beclin-1, VPS34 and LC3B were significantly increased (<0.01); the protein expressions of AMPK, Beclin-1, VPS34 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ were increased (<0.01, <0.05). After intervention and in comparison with the model group, the neurological function scores were decreased (<0.05); the cerebral infarct volume were decreased (<0.05); the contents of IL-6, IL-18 and TNF-α in cerebrospinal fluid were decreased (<0.05); the mRNA expressions of AMPK, Beclin-1, VPS34 and LC3B were significantly decreased (<0.01); the protein expressions of AMPK, Beclin-1, VPS34 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ were decreased (<0.05, <0.01).

CONCLUSION

EA can improve the neurological function and alleviate the degree of nerve injury in rats with cerebral I/R injury, which may be related to inhibiting the autophagy level of hippocampal neurons.

摘要

目的

探讨针刺“水沟”（GV6）、“百会”（GV20）对脑缺血再灌注（I/R）损伤大鼠海马神经元自噬的影响。

方法

将48只健康雄性SD大鼠随机分为假手术组、模型组和电针组，每组16只。采用大脑中动脉闭塞（MCAO）法制备大鼠脑I/R损伤模型。电针组大鼠于GV26和GV20行电针治疗20分钟，每日1次，连续5天。采用Longa神经功能评分法评估各组大鼠神经功能。通过TTC染色测量脑梗死体积。采用ELISA法检测脑脊液中IL-6、IL-18和TNF-α水平。分别采用实时荧光定量PCR和蛋白质免疫印迹法检测自噬相关蛋白AMPK、Beclin-1、VPS34和LC3B的表达。

结果

与假手术组比较，模型组大鼠神经功能评分显著升高（<0.01）；脑梗死体积显著增大（<0.01）；脑脊液中IL-6、IL-18和TNF-α含量升高（<0.01，<0.05）；AMPK、Beclin-1、VPS34和LC3B的mRNA表达水平显著升高（<0.01）；AMPK、Beclin-1、VPS34蛋白表达及LC3B-Ⅱ/LC3B-Ⅰ比值升高（<0.01，<0.05）。干预后与模型组比较，神经功能评分降低（<0.05）；脑梗死体积减小（<0.05）；脑脊液中IL-6、IL-18和TNF-α含量降低（<0.05）；AMPK、Beclin-1、VPS34和LC3B的mRNA表达显著降低（<0.01）；AMPK、Beclin-1、VPS34蛋白表达及LC3B-Ⅱ/LC3B-Ⅰ比值降低（<0.05，<0.01）。