康复训练对缺血性脑卒中小鼠突触蛋白Ⅰ和突触后致密蛋白-95表达的影响
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摘要
目的:探究局部缺血性脑卒中后康复训练对突触蛋白Ⅰ(SynapsinⅠ)和突触后致密蛋白-95(PSD-95)表达的影响。方法:光栓法建立局部缺血脑卒中模型,随机分为假手术组(n=9)、缺血组(n=9)、缺血+训练组(n=9),术后16h开始对缺血+训练组进行康复训练,持续至21天,分别于术后第1、3、7、14、21天对小鼠进行行为学测试。康复训练的方法有加速转棒训练、爬梯子训练、平衡木训练、悬挂训练,2次/d。21天后取脑做Western Blot分析SynapsinⅠ、PSD-95的表达水平。结果:行为测试结果表明,脑卒中后第1天缺血组和缺血+训练组疲劳转棒的成绩和抓力显著低于假手术组(P<0.01),在训练后第3天,缺血+训练组转棒成绩增高至假手术组水平,均明显高于缺血组(P<0.05),在训练第7天后,转棒成绩三组间无差异;前肢抓力恢复较慢,训练第7天后,缺血+训练组抓力提高至假手术组水平,较缺血组高(P<0.05)。Western Blot结果显示,术后21天,缺血组PSD-95表达水平明显低于假手术组(P<0.01),缺血+训练组显著高于缺血组(P<0.01);SynapsinⅠ表达水平缺血组较假手术组低(P<0.05),缺血+训练组显著高于缺血组(P<0.01)。结论:局部缺血性脑卒中后康复训练促进小鼠脑功能恢复与突触蛋白SynapsinⅠ和PSD-95表达的增加有关。
Objective: To explore the effect of functional recovery of rehabilitation training and its mechanism following ischemic stroke in mice.Method: Ischemic stroke was induced by photothrombosis. All mice were randomly divided into three groups:sham group, ischemia group and Ischemia + training group. In training group, the training was performed from 16 h after operation and lasted for 21 days. The methods of rehabilitation training include accelerated rotarod training, climb ladder training, balance beam training and hanging wire training twice a day. Behavioral tests were given at 1, 3, 7, 14, and 21 days. All animals were decapitated at 21 days. The expression of PSD-95 and Synapsin Ⅰ were detected by Western Blot.Result: Rotarod performance and limb grip in ischemia group and ischemia+training group were reduced significantly compared to sham group at 1 day after stroke(P<0.01). Compared to ischemia group, rotarod performance in ischemia + training group was raised to the sham group level at 3 days following stroke(P<0.05).There were no significance among three groups from 7 days after stroke. Limb grip restored more slowly. Grip increased to sham group level after 7 days training with significant difference with ischemia group(P<0.05).Western Blot analysis revealed the level of PSD-95 and Synapsin Ⅰcompared to sham group(P<0.01, P<0.05,respectively). However, both of them were increased significantly after training(P<0.01).Conclusion: Increased SynapsinⅠ and PSD-95 may involve in promoting functional recovery in ischemic stroke mice.
Objective: To explore the effect of functional recovery of rehabilitation training and its mechanism following ischemic stroke in mice.Method: Ischemic stroke was induced by photothrombosis. All mice were randomly divided into three groups:sham group, ischemia group and Ischemia + training group. In training group, the training was performed from 16 h after operation and lasted for 21 days. The methods of rehabilitation training include accelerated rotarod training, climb ladder training, balance beam training and hanging wire training twice a day. Behavioral tests were given at 1, 3, 7, 14, and 21 days. All animals were decapitated at 21 days. The expression of PSD-95 and Synapsin Ⅰ were detected by Western Blot.Result: Rotarod performance and limb grip in ischemia group and ischemia+training group were reduced significantly compared to sham group at 1 day after stroke(P<0.01). Compared to ischemia group, rotarod performance in ischemia + training group was raised to the sham group level at 3 days following stroke(P<0.05).There were no significance among three groups from 7 days after stroke. Limb grip restored more slowly. Grip increased to sham group level after 7 days training with significant difference with ischemia group(P<0.05).Western Blot analysis revealed the level of PSD-95 and Synapsin Ⅰcompared to sham group(P<0.01, P<0.05,respectively). However, both of them were increased significantly after training(P<0.01).Conclusion: Increased SynapsinⅠ and PSD-95 may involve in promoting functional recovery in ischemic stroke mice.
引文
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[25]Briones TL,Suh E,Jozsa L,et al.Changes in number of synapses and mitochondria in presynaptic terminals in the dentate gyrus following cerebral ischemia and rehabilitation training[J].Brain Research,2005,1033(1):51-57.
[26]Greengard P,Valtorta F,Czernik AJ,et al.Synaptic vesicle phosphoproteins and regulation of synaptic function[J]Science,1993,259(5096):780.
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[28]郭全虎.超早期推拿对骨骼肌急性钝挫伤模型大鼠膜修复蛋白Dysferlin表达的影响[D].重庆医科大学,2017.
[29]Kornau HC,Schenker LT,Kennedy MB,et al.A domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95[J].Science,1995,269(5231):1737.
[2]宇传华,罗丽莎,李梅,等.从全球视角看中国脑卒中疾病负担的严峻性[J].公共卫生与预防医学,2016,27(1):1-5.
[3]曹江慧,刘广志,周军,等.脑微出血对急性脑梗死患者溶栓治疗后出血转化及功能预后的影响:一项Meta分析[J].神经损伤与功能重建,2015,(5):399-402.
[4]姚敏,吴江.急性脑梗死溶栓治疗的研究与展望[J].脑卒中与神经疾病,2007,24(1):123-125.
[5]Pagnussat AS,Simao F,Anastacio JR,et al.Effects of skilled and unskilled training on functional recovery and brain plasticity after focal ischemia in adult rats[J].Brain Res,2012,1486:53-61.
[6]Nie J,Yang X.Modulation of synaptic plasticity by exercise training as a basis for ischemic stroke rehabilitation[J].Cellular&Molecular Neurobiology,2017,37(1):5-16.
[7]Kim G,Kim E.The effects of antecedent exercise on motor function recovery and brain-derived neurotrophic factor expression after focal cerebral ischemia in rats[J].Journal of Physical Therapy Science,2013,25(5):553.
[8]Xing Y,Yang SD,Dong F,et al.The beneficial role of early exercise training following stroke and possible mechanisms[J].Life Sciences,2018,198:32-37.
[9]徐振华.针刺不同刺激量对脑缺血后功能恢复及突触可塑性促进作用的研究[J].广州中医药大学,2005.
[10]王海潮.Calpain活性与PSD-95在AD模型小鼠脑组织中的表达差异研究[J].北京:北京交通大学,2016.
[11]汪江碧.孕酮联合康复训练对小鼠缺血性脑卒中的影响及机制研究[J].兰州大学,2013.
[12]Wang J,Feng X,Du Y,et al.Combination treatment with progesterone and rehabilitation training further promotes behavioral recovery after acute ischemic stroke in mice[J].Restorative Neurology&Neuroscience,2013,31(4):487-499.
[13]李红玲,赵巧艳,王红莲,等.脑卒中实验动物的行为学检测方法[J].中国康复医学杂志,2006,21(6):560-562.
[14]Wong R,Ray D,Kendall DA.Progesterone pharmacokinetics in the mouse:implications for potential stroke therapy[J].Journal of Pharmacy&Pharmacology,2012,64(11):1614-1620.
[15]Brown AW,Schultz BA.Recovery and rehabilitation after stroke[J].Seminars in Neurology,2010,30(05):511-517.
[16]Yang YR,Wang RY,Wang PS.Early and late treadmill training after focal brain ischemia in rats[J].Neuroscience Letters,2003,339(2):91-94.
[17]Ploughman M,Attwood Z,White N,et al.Endurance exercise facilitates relearning of forelimb motor skill after focal ischemia[J].European Journal of Neuroscience,2007,25(11):3453-3460.
[18]赵强,朱伟杰,王芙昱,等.小鼠局灶性脑缺血模型的比较分析[J].解放军医学院学报,2014,35(8):870-872.
[19]Pan R,Cai J,Zhan L,et al.Buyang Huanwu decoction facilitates neurorehabilitation through an improvement of synaptic plasticity in cerebral ischemic rats[J].Bmc Complementary&Alternative Medicine,2017,17(1):173.
[20]Shi N,Zhu C,Li L.Rehabilitation training and resveratrol improve the recovery of neurological and motor function in rats after cerebral ischemic injury through the sirt1 signaling pathway[J].Biomed Res Int,2016,2016:1732163.
[21]Buitrago MM,Schulz JB,Dichgans J,et al.Short and long-term motor skill learning in an accelerated rotarod training paradigm[J].Neurobiology of Learning&Memory,2004,81(3):211-216.
[22]Matsuda F,Sakakima H,Yoshihiro Y.The effects of early exercise on brain damage and recovery after focal cerebral infarction in rats[J].Neuroscience Research,2010,65(2):S101-S101.
[23]徐莉,李玲,陈景藻.康复训练对大鼠脑梗死神经功能恢复的影响[J].中华物理医学与康复杂志,2000,(2):86-88.
[24]Nudo RJ,Friel KM.Cortical plasticity after stroke:implications for rehabilitation[J].Revue Neurologique,1999,155(9):713-717.
[25]Briones TL,Suh E,Jozsa L,et al.Changes in number of synapses and mitochondria in presynaptic terminals in the dentate gyrus following cerebral ischemia and rehabilitation training[J].Brain Research,2005,1033(1):51-57.
[26]Greengard P,Valtorta F,Czernik AJ,et al.Synaptic vesicle phosphoproteins and regulation of synaptic function[J]Science,1993,259(5096):780.
[27]Kleim JA,Lussnig E,Schwarz ER,et al.Synaptogenesis and Fos expression in the motor cortex of the adult rat after motor skill learning[J].J Neurosci,1996,16(14):4529-4535.
[28]郭全虎.超早期推拿对骨骼肌急性钝挫伤模型大鼠膜修复蛋白Dysferlin表达的影响[D].重庆医科大学,2017.
[29]Kornau HC,Schenker LT,Kennedy MB,et al.A domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95[J].Science,1995,269(5231):1737.