中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (9): 1594-1601.doi: 10.4103/1673-5374.235294

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

电针干预JNK基因敲除小鼠脑缺血再灌注损伤：如何发挥神经保护作用

  

  • 收稿日期:2018-04-02 出版日期:2018-09-15 发布日期:2018-09-15
  • 基金资助:

    中国国家自然科学基金项目（81173355）

Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism

Chun-Xiao Wu1, 2, Yi-Hui Feng3, Lu Yang1, 4, Zhu-Lian Zhan4, Xiu-Hong Xu4, Xiao-Ying Hu4, Zhi-Hua Zhu5, Guo-Ping Zhou1, 4   

  1. 1 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong Province, China
    2 Medical College of Acu-Moxi and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China
    3 Central Hospital of Qingdao, Qingdao, Shandong Province, China
    4 Traditional Chinese Medicine-Integrated Hospital, the Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong Province,China
    5 Ruikang Hospital Affiliated to Guangxi College of Traditional Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China
  • Received:2018-04-02 Online:2018-09-15 Published:2018-09-15
  • Contact: Guo-Ping Zhou, M.D.,doctorzgp@sina.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81173355.

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摘要：

单纯调节JNK或p38 MAPK通路不足以触发细胞凋亡的过程，而激活应激激活通路（JNK/p38 MAPK）和抑制生长因子活化的ERK通路才能促进细胞凋亡。为此，作者假设抑制JNK或p38可促细胞凋亡发生，同时促进ERK通路可能是脑内缺血再灌注损伤的抗凋亡机制。为明确电针干预JNK基因敲除小鼠脑缺血再灌注损伤的作用机制，实验以Longa法建立JNK基因敲除小鼠局灶性脑缺血再灌注损伤模型，在损伤后1.5h电针刺激尺泽(LU5)、合谷(LI4)、三阴交(SP6)和足三里(ST36)，深度2-3mm，频率5/10Hz，强度2mA，持续时间20min，1次/d。以神经功能缺损评分评价神经功能，用双标免疫荧光和Western blot检测损伤侧大脑皮质和海马中p-ERK和p-p38的免疫阳性反应及相对表达水平，以实时定量PCR检测损伤侧大脑皮质和海马中ERK和p38 mRNA的表达。发现经电针治疗后，脑缺血再灌注损伤小鼠的神经功能改善，损伤侧大脑皮质和海马中p-ERK免疫阳性反应及相对表达增加， p-p38免疫阳性反应及相对表达减少，且损伤侧大脑皮质和海马中ERKmRNA表达增加，而p38mRNA表达减少。数据结果提示电针上调ERK保护通路，同时抑制了p38的凋亡通路，从而发挥了神经保护作用，改善了JNK基因敲除小鼠的神经功能。

orcid:0000-0002-4613-4384(Guo-Ping Zhou)

 

关键词: MAPK信号通路, 脑缺血再灌注, 应激激活通路, 生长因子激活通路, 电针, 穴位, 细胞凋亡, 大脑皮质, 海马, 双标记免疫荧光, 共聚焦激光扫描显微镜, Western blot分析, 定量PCR, JNK基因敲除小鼠

Abstract:

Simple regulation of c-Jun N-terminal kinase (JNK) or p38 mitogen-activated protein kinase (MAPK) pathways is not enough to trigger cell apoptosis. However, activation of the stress activated pathway (JNK/p38 MAPK) together with inhibition of the growth factor activated extracellular signal-regulated kinase (ERK) pathway can promote cell apoptosis. We hypothesized that inhibition of the JNK or p38 pro-apoptotic pathway and activating the ERK pathway could be the mechanism of anti-apoptosis following cerebral ischemia/reperfusion injury. To investigate the mechanism of the protective effect of electroacupuncture on cerebral ischemia/reperfusion injury in JNK knockout mice, mouse models of cerebral ischemia/reperfusion injury were established by Longa’s method. Electroacupuncture was conducted at acupoints Chize (LU5), Hegu (LI4), Sanyinjiao (SP6) and Zusanli (ST36) 1.5 hours after ischemia/reperfusion injury for 20 minutes,once a day. The neurological function was evaluated using neurological deficit scores. The expression of phospho-extracellular signal-regulated kinase (p-ERK) and phospho-p38 (p-p38) in JNK knockout mice was detected using double-labeling immunofluorescence and western blot assay. The mRNA expression of ERK and p38 was measured by quantitative real-time polymerase chain reaction. Electroacupuncture improved neurological function, increased the immunoreactivity and relative expression of p-ERK and reduced that of p-p38 in the cerebral cortex and hippocampus on the injured side. Electroacupuncture increased mRNA expression of ERK, but decreased that of p38 in the cerebral cortex and hippocampus on the injured side. In conclusion, electroacupuncture upregulated the protective ERK pathway and inhibited the pro-apoptotic p38 pathway, thereby exerting a neuroprotective effect and improving the neurological function in JNK knockout mice.

Key words: nerve regeneration, MAPKs signaling pathway, cerebral ischemia/reperfusion, stress-activated pathway, growth factor-activated pathway, electroacupuncture, apoptosis