中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (5): 775-782.doi: 10.4103/1673-5374.249225

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

缺血性脑损伤大鼠接受早期强制性运动疗法对行为和神经元可塑性的变化

  

  • 出版日期:2019-05-15 发布日期:2019-05-15
  • 基金资助:

    山东省自然科学基金(2014ZRB14502)

Early constraint-induced movement therapy affects behavior and neuronal plasticity in ischemia-injured rat brains

Xi-Hua Liu 1 , Hong-Yan Bi 1 , Jie Cao 2 , Shuo Ren 1 , Shou-Wei Yue 3   

  1. 1 Department of Physical Medicine & Rehabilitation, Affiliated Hospital of Shandong Traditional Chinese Medicine University, Jinan, Shandong Province, China
    2 Maternal and Child Health Development Research Center, Shandong Maternal and Child Health Hospital, Jinan, Shandong Province, China
    3 Department of Physical Medicine & Rehabilitation, Qilu Hospital, Shandong University, Jinan, Shandong Province, China
  • Online:2019-05-15 Published:2019-05-15
  • Contact: Shou-Wei Yue, MD, shouweiyue1224@163.com.
  • Supported by:

    This study was supported by the Natural Science Foundation of Shandong Province of China, No. 2014ZRB14502 (to XHL).

摘要:

强制性运动疗法(CIMT)是一种有效改善脑卒中后上肢运动功能恢复的康复训练技术,然而何时介入CIMT治疗会比常规康复治疗具有更好的疗效目前仍存在争议。为此,试验拟探讨缺血性脑损伤后CIMT的介入最佳时机,及促进脑卒中功能恢复的机制。将64只成年雄性SD大鼠随机分为4组:假手术组、模型组、E-CIMT组和L-CIMT组。模型组采用Longa线栓法建立大鼠左侧大脑中动脉闭塞模型,E-CIMT组和L-CIMT组分别于建模后1,14 d 开始介入强制性运动疗法。为了解不同介入时机对神经运动功能的影响,分别在建模前,建模后8,21 d采用平衡木试验进行神经功能评估;分别应用real time -PCR和Western blot检测脑源性神经营养因子、神经生长因子和Nogo蛋白受体基因和蛋白的表达,以了解不同介入时机对相关神经因子表达的影响。结果显示,(1)在建模后8 d,E-CIMT组的平衡木试验神经功能评分显著低于模型组和L-CIMT 组;在建模后21 d,E-CIMT组和L-CIMT 组的神经功能评分均显著降低;(2)建模后8 d,与模型组和L-CIMT组比较,E-CIMT组脑源性神经营养因子和神经生长因子mRNA及蛋白的表达明显升高,而Nogo蛋白受体 mRNA及蛋白的表达表达明显降低;(3)上述结果证实,脑梗死后早期(1 d)和晚期(14 d)介入CIMT均能诱导神经元可塑性,促进脑卒中后的功能恢复,其机制可能与下调Nogo蛋白受体表达,以及上调脑源性神经营养因子和神经生长因子的表达有关。

orcid: 0000-0002-1285-0702 (Shou-Wei Yue)

关键词: 缺血性脑卒中, 康复, 强制性运动疗法, 神经生长因子, 功能恢复, 神经元可塑性, 反转录聚合酶链反应, 免疫印迹试验, 大鼠, 神经再生

Abstract:

Constraint-induced movement therapy is an effective rehabilitative training technique used to improve the restoration of impaired upper extremity movement after stroke. However, whether constraint-induced movement therapy is more effective than conventional rehabilitation in acute or sub-acute stroke remains controversial. The aim of the present study was to identify the optimal time to start constraint-induced movement therapy after ischemic stroke and to explore the mechanisms by which constraint-induced movement therapy leads to post-stroke recovery. Sixty-four adult male Sprague-Dawley rats were randomly divided into four groups: sham-surgery group, cerebral ischemia/reperfusion group, early constraint-induced movement therapy group, and late constraint-induced movement therapy group. Rat models of left middle cerebral artery occlusion were established according to the Zea Longa line embolism method. Constraint-induced movement therapy was conducted starting on day 1 or day 14 in the early constraint-induced movement therapy and late constraint-induced movement therapy groups, respectively. To explore the effect of each intervention time on neuromotor function, behavioral function was assessed using a balance beam walking test before surgery and at 8 and 21 days after surgery. The expression levels of brain-derived neurotrophic factor, nerve growth factor and Nogo receptor were evaluated using real time-polymerase chain reaction and western blot assay to assess the effect of each intervention time. The results showed that the behavioral score was significantly lower in the early constraint-induced movement therapy group than in the cerebral ischemia/reperfusion and late constraint-induced movement therapy groups at 8 days. At 21 days, the scores had significantly decreased in the early constraint-induced movement therapy and late constraint-induced movement therapy groups. At 8 days, only mild pyknosis appeared in neurons of the ischemic penumbra in the early constraint-induced movement therapy group, which was distinctly better than in the cerebral ischemia/reperfusion group. At 21 days, only a few vacuolated cells were observed and no obvious inflammatory cells were visible in late constraint-induced movement therapy group, which was much better than at 8 days. The mRNA and protein expression levels of brain-derived neurotrophic factor and nerve growth factor were significantly higher, but expression levels of Nogo receptor were significantly lower in the early constraint-induced movement therapy group compared with the cerebral ischemia/reperfusion and late constraint-induced movement therapy groups at 8 days. The changes in expression levels at 21 days were larger but similar in both the early constraint-induced movement therapy and late constraint-induced movement therapy groups. Besides, the protein nerve growth factor level was higher in the late constraint-induced movement therapy group than in the early constraint-induced movement therapy group at 21 days. These results suggest that both early (1 day) and late (14 days) constraint-induced movement therapy induces molecular plasticity and facilitates functional recovery after ischemic stroke, as illustrated by the histology. The mechanism may be associated with downregulation of Nogo receptor expression and upregulation of brain-derived neurotrophic factor and nerve growth factor expression.

Key words: nerve regeneration, ischemic stroke, rehabilitation, constraint-induced movement therapy, nerve growth factors, functional recovery, neuronal plasticity, real time-polymerase chain reaction, western blot assay, rats, neural regeneration