摘要
目的观察不同强度运动训练对脑缺血再灌注大鼠脑内臂板蛋白3A（Sema3A）及其受体神经纤毛蛋白-1（NP．1）的表达以及缺血侧脑细胞凋亡的影响，探讨强化运动训练促进脑缺血再灌注大鼠运动功能恢复的可能机制。方法采用线栓法建立左侧大脑中动脉阻塞（MCAO）2h再灌注动物模型。60只造模成功的雄性Wistar大鼠按随机数字表法分为训练1组、训练2组、训练3组、对照组（各15只鼠）。训练1组大鼠每天游泳1次，每次5min；训练2组大鼠每天游泳1次，每次10min；训练3组大鼠每天游泳2次，每次10min；对照组大鼠不做任何训练；另15只鼠为假手术组不阻塞大脑中动脉血流，不训练。采用Garcia神经功能缺损评分评价神经功能缺损情况。采用TUNEL染色观察缺血区大脑皮质细胞凋亡情况。应用免疫组织化学法对脑缺血后脑内神经生长抑制因子Sema3A及其受体NP-1的表达进行分析。结果假手术组神经行为功能正常。对照组各时间点的Garcia神经行为功能评分与假手术组同时间点比较，差异均有统计学意义（P〈0．01）；各训练组游泳训练7和14d后的Garcia神经功能评分明显优于同时间点对照组（P〈0．01），且训练3组神经功能评分提高最明显，其游泳训练3、7和14d后的Garcia神经功能评分分别为（12．80±0．45）、（15．204-0．45）和（16．80-4-0．45）分。游泳训练3、7和14d后，训练1组、训练2组、训练3组的Sema3A、NP-l及TUNEL各指标阳性细胞的表达均明显低于对照组相同时间点（P〈0．01），以训练3组更为明显。训练3组游泳训练3、7和14d后，TUNEL阳性细胞率分别为（29．43±1．38）％、（22．30±1．21）％和（17．58±1．70）％；Sema3A阳性细胞率分别为（19．64±1．17）％、（9．73-4-3．83）％和（8．24-t-O．87）％；NP-1阳性细胞率分别为（33．95-4-6．86）％、（27．95±1．29）％和（18．90±1．44）％，较其他各训练组的阳性细胞表达减少更明显（P〈0．01或0．05）。结论康复训练可减少脑缺血再灌注大鼠Sema3A、NP-1及TUNEL阳性细胞表达，促进其运动功能的恢复及神经功能重塑，强化运动训练的效果更明显。
[Abstract] Objective To observe the effects of intensive training at different intensities on the expressions of semaphorin 3A （ Sema 3A） and its receptor neuropilin （ NP-1 ） and the cell apoptosis in cerebrum after cerebral is- cbemia-reperfusion in rats, and to investigate the possible mechanism of intensive training in recovery of motor func- tion after cerebral ischemia-reperfusion in rats. Methods To establish animal model of cerebral ischemia-reperfu- sion in rats, the intraluminal thread method was applied to cause left middle cerebral artery occlusion （MCAO） for 2 h and before reperfusion. After cerebral ischemia-reperfusion model were established for 24 h, 60 male model Wist- ar rats were randomly divided into training group I （ swimming for 5 min once a day） , training group 2 （ swimming for 10 min once a day） , training group 3 （swimming for 10 rain twice a day） and control group （no training） ; another 15 rats assigned to the sham-operation group were subject to no MCAO and no training. Neurological function was e- valuated by Garcia scores, and terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling （TUNEL） was performed to dectect the cortical cell apoptosis. Expressions of neural growth inhibition factor Sema 3A and its receptor NP-1 were detected by immunohistochemistry. Results The neurological function in sham-operation group was normal. The differences of Garcia scores at different time points beween sham-operationgroup and control group were significant （P 〈 0.01 ）. Garcia scores in all training groups, were significantly higher than those in controls at the 7th and ldth d after swimming training （P 〈 O. O1 ）, especially in training group 3 the Garcia scores were （ 12.80 ±O. 45 ）, （ 15.20 ± 0.45 ）, （ 16.80 ±0.45 ）, respectively, at the 3rd, 7th and 14th d af- ter swimming training. The rates of positive cell of Sema 3A, NP-1 and TUNEL indexes in all training groups were lower than those in controls at the 3rd, 7th and 14th d after swimming training （P 〈 O. 01 ） , especially in the training group 3. At the 3rd, 7th and 14th d after swimming training in training group 3, the rates of TUNEL indexes positive apoptosis cells were （29.43 ± 1.38） % , （22.30 ± 1.21 ） % , （ 17.58± 1.70） % , respectively, the positive cell rates of Sema 3A were （19.64 ± 1. 17） % , （9.73 ± 3.83） % , （8.24 ± 0.87）% , respectively, the positive cell rates of NP-1 were （ 33.95 ± 6.86） %, （ 27.95 ± 1.29 ） %, （ 18.90 ± 1.44 ） %, respectively, the reduction of positive cells expressions in training group 3 was significantly more obvious compared with other training groups （P 〈 0.01 or 0.05）. Conclusions Rehabilitation training can reduce the expression of positive cell of Sema 3A, NP-1 and TUNEL indexes in rats after cerebral ischemia-reperfusion and can improve motor function recovery and facilitate neu- ral olasticitv. The more intensive the traininz, the better the effects.
出处
《中华物理医学与康复杂志》
CAS
CSCD
北大核心
2012年第1期2-7,共6页
Chinese Journal of Physical Medicine and Rehabilitation
基金
山东省教育厅科研发展计划（J09LF22）
