强制性运动疗法刺激脑缺血模型大鼠残存感觉运动皮质并抑制CRMP2磷酸化

doi:10.3877/cma.j.issn.1673-9248.2025.05.010

目的

通过探索强制性运动疗法（CIMT）对脑卒中后大脑双侧皮质神经元激活模式及塌陷反应介质蛋白2（CRMP2）磷酸化的影响，解析CIMT改善脑卒中后运动功能的作用机制。

方法

采用“线栓法”建立左侧大脑中动脉闭塞（MCAO）大鼠模型，随机分为CIMT组（n=7）或对照组（n=6）。自模型建立后第8天开始给予CIMT组为期2周的CIMT，对照组笼内自由活动。分别在术前（D0）及术后第7天（D7）、第14天（D14）和第21天（D21）使用Footfault评估系统评估大鼠右前肢的运动功能；在MCAO术后第16、21天，通过蛋白质印迹实验检测双侧感觉运动皮质中ΔFosB与CRMP2的含量，分析CIMT过程中左、右脑的激活模式，观察CIMT对CRMP2和CRMP2磷酸化（p-CRMP2）的影响。使用两独立样本t检验（当方差齐时）或校正t检验（当方差不齐时）进行组间差异比较。

结果

在术后D14和D21时，CIMT组的Footfault有效放置率均显著高于对照组（0.43±0.05 vs 0.30±0.06；0.56±0.08 vs 0.31±0.07），差异均有统计学意义（t=3.971、6.161，P=0.002、<0.001）。MCAO术后第16天，CIMT组左侧（梗死侧）感觉运动皮质的ΔFosB含量显著高于对照组（0.064±0.009 vs 0.036±0.004），差异有统计学意义（t=2.947，P=0.026）。MCAO术后第21天，CIMT组左侧（梗死侧）大脑感觉运动皮质的p-CRMP2含量及p-CRMP2/CRMP2含量比值均显著低于对照组（0.371±0.095 vs 0.786±0.096；0.263±0.063 vs 0.567±0.094），差异均有统计学意义（t=2.987、2.792，P=0.014，0.019）。

结论

脑卒中后CIMT直接刺激梗死侧残存感觉运动皮质，抑制梗死侧感觉运动皮质中CRMP2的磷酸化，提示梗死侧残存感觉运动皮质在神经重塑中具有重要的作用。

关键词: 强制性运动疗法, 脑缺血, 蛋白质, 运动皮质, 康复

Objective

To elucidate the mechanism of constraint-induced movement therapy (CIMT) improves motor function after stroke by examining its effects of CIMT on the bilateral cortical neurons activation patterns and collapsin response mediator protein 2 phosphorylation (CRMP2).

Methods

In this study, a left middle cerebral artery occlusion (MCAO) rat model was established using the "thread embolization method", and the rats were randomly divided into a CIMT group (n=7) or a control group (n=6). Starting on day 8 after modeling, the CIMT group underwent 2 weeks of CIMT training, while the control group was allowed to move freely in the cage. The motor function of the right forelimb was evaluated using the Footfault assessment system at baseline (D0), day 7 (D7), day 14 (D14), and day 21 (D21) post-surgery. On days 16 and 21 after MCAO, the contents of ΔFosB and CRMP2 in the bilateral sensorimotor cortex were detected to analyze activation patterns of the left and right brain during CIMT and evalute the effect of CIMT on CRMP2 and phosphorylated CRMP2 (p-CRMP2). Group comparisons were performed using independent samples t-test (for homogeneous variances) or Welch's corrected t-test (for heterogeneous variances).

Results

The Footfault effective placement rates were significantly higher in the CIMT group than in the control group at D14 (0.43±0.05 vs 0.30±0.06, t=3.971, P=0.002) and D21 (0.56±0.08 vs 0.31±0.07, t=6.161, P<0.001). On day 16 post-MCAO, the ΔFosB content in the left (infarcted side) sensorimotor cortex of the CIMT group was significantly higher than that of the control group (0.064±0.009 vs 0.036±0.004, t=2.947, P=0.026). On day 21, both p-CRMP2 levels and the p-CRMP2/CRMP2 ratio in the infarcted sensorimotor cortex were significantly lower in the CIMT group (0.371±0.095 vs 0.786±0.096, t=2.987, P=0.014; 0.263±0.063 vs 0.567±0.094, t=2.792, P=0.019).

Conclusion

CIMT enhances activation of the residual ipsilesional cortex and suppresses CRMP2 phosphorylation in the ischemic hemisphere suggesting that the residual cortex on the affected side plays an important role in neural remodeling during post-stroke recovery.

Key words: Constraint-induced movement therapy, Ischemic stroke, Protein, Residual cortex, Rehabilitation

表1 大脑中动脉闭塞模型大鼠Footfault运动有效放置率比较（

x ¯

±s）

组别	只数	术前	术后第7天	术后第14天	术后第21天
CIMT组	7	0.84±0.06	0.28±0.06	0.43±0.05^a	0.56±0.08^ab
对照组	6	0.79±0.06	0.25±0.07	0.30±0.06	0.31±0.07
t值		1.309	0.838	3.971	6.161
P值		0.217	0.416	0.002	<0.001

表1 大脑中动脉闭塞模型大鼠Footfault运动有效放置率比较（

x ¯

±s）

组别	只数	术前	术后第7天	术后第14天	术后第21天
CIMT组	7	0.84±0.06	0.28±0.06	0.43±0.05^a	0.56±0.08^ab
对照组	6	0.79±0.06	0.25±0.07	0.30±0.06	0.31±0.07
t值		1.309	0.838	3.971	6.161
P值		0.217	0.416	0.002	<0.001

图1 大脑中动脉闭塞模型建立第16天（D16）后大鼠双侧感觉运动皮质ΔFosB含量检测注：CIMT为强制性运动疗法；Control为对照；L为左侧感觉运动皮质；R为右侧感觉运动皮质；GAPDH为甘油醛-3-磷酸脱氢酶；β-actin和GAPDH为内参蛋白

图2 大脑中动脉闭塞模型建立第21天（D21）后大鼠大脑感觉运动皮质塌陷反应介质蛋白2磷酸化（p-CRMP2）和p-CRMP2/塌陷反应介质蛋白2（CRMP2）比值比较注：CIMT为强制性运动疗法；Control为对照；R为右侧；L为左侧

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Constraint-induced movement therapy stimulates the residual sensorimotor cortex and inhibits the phosphorylation of CRMP2 in a rat model of cerebral ischemia

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Constraint-induced movement therapy stimulates the residual sensorimotor cortex and inhibits the phosphorylation of CRMP2 in a rat model of cerebral ischemia