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高蓓瑶, 许东升, 谢蓓菁, 等. 改良强制性运动调节脑缺血再灌注模型大鼠运动皮层内神经递质水平的研究[J]. 四川大学学报(医学版), 2020, 51(5): 611-617. DOI: 10.12182/20200960601
引用本文: 高蓓瑶, 许东升, 谢蓓菁, 等. 改良强制性运动调节脑缺血再灌注模型大鼠运动皮层内神经递质水平的研究[J]. 四川大学学报(医学版), 2020, 51(5): 611-617. DOI: 10.12182/20200960601
GAO Bei-yao, XU Dong-sheng, XIE Bei-jing, et al. Effect of Modified Constraint-induced Movement Therapy on Neurotransmitter Levels of Motor Cortex in Cerebral Ischemia-Reperfusion Injured Rats[J]. Journal of Sichuan University (Medical Sciences), 2020, 51(5): 611-617. DOI: 10.12182/20200960601
Citation: GAO Bei-yao, XU Dong-sheng, XIE Bei-jing, et al. Effect of Modified Constraint-induced Movement Therapy on Neurotransmitter Levels of Motor Cortex in Cerebral Ischemia-Reperfusion Injured Rats[J]. Journal of Sichuan University (Medical Sciences), 2020, 51(5): 611-617. DOI: 10.12182/20200960601

改良强制性运动调节脑缺血再灌注模型大鼠运动皮层内神经递质水平的研究

Effect of Modified Constraint-induced Movement Therapy on Neurotransmitter Levels of Motor Cortex in Cerebral Ischemia-Reperfusion Injured Rats

  • 摘要:
      目的  研究改良强制性运动对脑缺血再灌注模型大鼠运动功能恢复的作用及其机制。
      方法  将大鼠随机分为对照组和改良强制性运动疗法(mCIMT)组,每组12只。采用Longa线栓法建立大鼠左侧大脑中动脉栓塞(MCAO)模型。mCIMT组在造模后7 d开始连续训练14 d,使用有弹性的的绷带将健肢绑于前胸,并在强制性转轮设备中进行患肢活动的训练,对照组大鼠不进行mCIMT训练,仅笼内自由活动。在造模后20 d内记录大鼠体质量,通过错步实验评定大鼠的行为学变化。在造模后18 d时处死部分大鼠,使用高效液相色谱法(high performance liquid chromatography, HPLC)检测脑运动皮层和纹状体内单胺类神经递质〔5-羟色胺(5-HT)、5-羟吲哚乙酸(5-HIVV)和高香草酸(HVA)〕和氨基酸类神经递质〔天门冬氨酸(ASP)、谷氨酰胺(Gln)、甘氨酸(Gly)、牛磺酸(Tau)、γ 氨基丁酸(GABA)〕含量,酶联免疫吸附(ELISA)法检测脑运动皮层和纹状体组织总p70核糖体蛋白S6激酶(p70s6k)蛋白和磷酸化蛋白(p-p70s6k)的表达水平。
      结果  造模后21 d内,与对照组比较,mCIMT组大鼠体质量无明显变化(P>0.05),mCIMT组大鼠在造模后17 d时错步率下降(P<0.05);造模 后18 d时,与对照组比较,mCIMT组大鼠运动皮层中单胺类神经递质5-HIVV水平增加(P<0.05),运动皮层中氨基酸类神经递质的相对含量(与Glu之比)除ASP/Glu降低(P<0.05)外,其余氨基酸类神经递质包括Gln、Gly、Tau、GABA与Glu之比均增加(P<0.05或 P<0.01)。与对照组比较,mCIMT组大鼠运动皮层中p-p70s6k表达减少(P<0.05)。而对照组与mCIMT组大鼠纹状体中的单胺类神经递质和氨基酸神经递质差异均无统计学意义(P>0.05)。
      结论  mCIMT具有一定的改善脑缺血再灌注模型大鼠运动功能的效果,其机制可能与提高大鼠脑运动皮层内氨基酸类神经递质和5-HIVV水平、减少p-p70s6k表达有关。

     

    Abstract:
      Objective  To study the effect and mechanism of modified constraint-induced movement therapy (mCIMT) on motor function recovery in cerebral ischemia-reperfusion rats.
      Methods  The rats were randomly divided into the control group and the mCIMT group, with 12 rats in each group. The left middle cerebral artery occlusion (MCAO) model was established by the Longa suture method. In the mCIMT group, the rats started continuous training for 14 d on the 7th day after modeling. The unaffected limb was tied to the chest with elastic bandages, and the affected limb was trained in the compulsory runner equipment. In the control group, rats moved freely in the cage. The body mass of rats was recorded within 20 d after modeling, and behavior was assessed by the foot-fault test. Some of the rats were euthanized 18 d after modeling, and high performance liquid chromatography (HPLC) was used to detect monoamine neurotransmitters (5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIVV), homovanillic acid (HVA) ), and amino acid neurotransmitters (glutamic acid (Glu), asparaginic acid (ASP), glutamine (Gln), glycine (Gly), taurine (Tau), gamma aminobutyric acid (GABA) ) in the motor cortex and striatum, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of total P70 ribosomal protein S6 kinase (p70s6k) and p70s6k phosphorylated protein (p-p70s6k) in motor cortex and striatum, respectively.
      Results  Compared with the control group, the body mass of rats in the mCIMT group was comparable (P>0.05) within 21 d after modeling, foot-fault rate of the mCIMT group was significantly lower at 17 d after modeling (P<0.05). At 18 d after modeling, compared with the control group, the level of 5-HIVV in the motor cortex increased significantly (P<0.05), and the relative content of amino acid neurotransmitters (the ratio of Glu) in the motor cortex including Gln, Gly, Tau and GABA to Glu increased significantly (P<0.05 or P<0.01) except for decreased ASP/Glu (P<0.05). Moreover, compared with the control group, the expression of p-p70s6k in the motor cortex of the mCIMT was significantly decreased (P<0.05). There were no significant differences in monoamine neurotransmitters and amino acid neurotransmitters in the striatum between two groups (P>0.05).
      Conclusion  mCIMT improved the motor function of MCAO rats, and the mechanism might be related to the increase of amino acid neurotransmitters and 5-HIVV and decrease of p-p70s6k expression in the motor cortex.

     

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