针刺结合疲劳转棒训练改善pMCAO大鼠运动和认知功能
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摘要
目的:探讨针刺靳三针头穴结合疲劳转棒康复训练对永久性大脑中动脉闭塞(permanent middle cerebral artery occlusion,pMCAO)大鼠运动功能和认知功能恢复的影响。方法:将38只雄性SD大鼠随机分为3组,假手术组11只,模型组13只,治疗组14只。采用电凝法建立右侧永久性大脑中动脉闭塞模型。于造模后第1天,治疗组针刺"智三针""颞三针""脑三针",并进行疲劳转棒训练,每日针1次,训练3次。假手术组及模型组仅在治疗组针刺时进行空抓固定,其余时刻不做其他处理。连续干预6 d休息1 d,共干预3周。造模后,每组于第1天各取3只进行脑切片TTC染色观察脑缺血情况,第1至7天进行神经功能评分,第1、7、14、21天进行爪印分析和疲劳转棒测试,第22至26天进行水迷宫实验。结果:与假手术组比较,模型组大鼠存在明显脑缺血,神经功能评分升高,左侧足心距离在第1、7、14天升高,疲劳转棒转速在4个时间点均下降,定位航行中潜伏期均升高,空间探索中原平台象限Q3的活动时间百分比下降(P<0.01,P<0.05)。与模型组比较,治疗组大鼠脑缺血面积差异不明显(P>0.05),神经功能评分在第6天下降,左侧足心距离在第14天下降,疲劳转棒转速在第14、21天升高,定位航行中潜伏期在第23至25天下降,空间探索中原平台象限Q3的活动时间百分比升高(P<0.01,P<0.05)。结论:针刺靳三针头穴结合疲劳转棒康复训练能够改善pMCAO大鼠的行为学表现,促进其运动功能和认知功能的恢复。
Objective To explore the effects on the recovery of the motor and cognitive functions of the rats with permanent middle cerebral artery occlusion(pMCAO) after treated with Jin's three-needle acupuncture at head acupoints combined with rota-rod training. Methods A total of 38 male SD rats were randomized into 3 groups, named a sham-operation group(11 rats), a model group(13 rats) and a treatment group(14 rats). The electrocoagulation method was adopted to establish the model of pMCAO on the right cerebrum. Starting from the 1 st day after successful modeling, acupuncture was applied to the "three points of intelligence", the "three points of temporal area" and the "three points of brain". Additionally, the rota-rod training was used. Acupuncture was given once a day and the training was three times a day. In the sham-operation group and the model group, empty grasp fixation was performed when acupuncture was applied in the treatment group, and there was no intervention at the rest of the time. There was 1 day of interval after consecutive 6 days of intervention. Totally, the intervention was for 3 weeks. After modeling, the brain section was collected from 3 rats of each group on the 1 st day and was stained with TTC to observe the condition of cerebral ischemia. From day 1 to 7, the neurological function score was evaluated. The footprint analysis and rota-rod test were performed on day 1, 7, 14 and 21. The Morris water maze test was performed from day 22 to 26. Results Compared with the sham-operation group, cerebral ischemia presented obviously, the score of neurological function was increased, the back front distances on the left were increased on day 1, 7 and 14 separately, the revolutions per minute(RPM) of the rota-rod were reduced at each of the above 4 time points, the latency of navigation trial was increased and the movement time percentage in Q3 quadrant of spatial probe trial was reduced in the model group(P<0.01, P<0.05). Compared with the model group, the area of cerebral ischemia was not obviously different(P>0.05), the score of neurological function was reduced on day 6, the back front distance on the left was reduced on day 14, RPM of the rota-rod were increased on day 14 and 21, the latency of navigation trial were reduced from day 23 to 25 and the movement time percentage in Q3 quadrant of spatial probe trial was increased in the treatment group(P<0.01, P<0.05). Conclusion Jin's three-needle acupuncture at head acupoints combined with rota-rod training improve the behavioral performance of pMCAO rats and promote the recovery of motor and cognitive functions.
Objective To explore the effects on the recovery of the motor and cognitive functions of the rats with permanent middle cerebral artery occlusion(pMCAO) after treated with Jin's three-needle acupuncture at head acupoints combined with rota-rod training. Methods A total of 38 male SD rats were randomized into 3 groups, named a sham-operation group(11 rats), a model group(13 rats) and a treatment group(14 rats). The electrocoagulation method was adopted to establish the model of pMCAO on the right cerebrum. Starting from the 1 st day after successful modeling, acupuncture was applied to the "three points of intelligence", the "three points of temporal area" and the "three points of brain". Additionally, the rota-rod training was used. Acupuncture was given once a day and the training was three times a day. In the sham-operation group and the model group, empty grasp fixation was performed when acupuncture was applied in the treatment group, and there was no intervention at the rest of the time. There was 1 day of interval after consecutive 6 days of intervention. Totally, the intervention was for 3 weeks. After modeling, the brain section was collected from 3 rats of each group on the 1 st day and was stained with TTC to observe the condition of cerebral ischemia. From day 1 to 7, the neurological function score was evaluated. The footprint analysis and rota-rod test were performed on day 1, 7, 14 and 21. The Morris water maze test was performed from day 22 to 26. Results Compared with the sham-operation group, cerebral ischemia presented obviously, the score of neurological function was increased, the back front distances on the left were increased on day 1, 7 and 14 separately, the revolutions per minute(RPM) of the rota-rod were reduced at each of the above 4 time points, the latency of navigation trial was increased and the movement time percentage in Q3 quadrant of spatial probe trial was reduced in the model group(P<0.01, P<0.05). Compared with the model group, the area of cerebral ischemia was not obviously different(P>0.05), the score of neurological function was reduced on day 6, the back front distance on the left was reduced on day 14, RPM of the rota-rod were increased on day 14 and 21, the latency of navigation trial were reduced from day 23 to 25 and the movement time percentage in Q3 quadrant of spatial probe trial was increased in the treatment group(P<0.01, P<0.05). Conclusion Jin's three-needle acupuncture at head acupoints combined with rota-rod training improve the behavioral performance of pMCAO rats and promote the recovery of motor and cognitive functions.
引文
[1]徐世芬,庄礼兴,贾超,等.“靳三针”对脑卒中偏瘫患者认知功能和日常生活能力的影响:多中心随机对照研究[J].中国针灸,2009,29(9):689-694.
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[6]葛亚博,冯晓东.电针疗法联合康复运动训练治疗脑卒中肢体痉挛大鼠的机制[J].中国老年学杂志,2017,37(15):3643-3645.
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[11]曹勇.针刺对HIBD模型大鼠远期行为学及Ca2+浓度、LDH活性的影响[D].广州:广州中医药大学,2014.
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[13]袁青.靳三针法[M].北京:人民卫生出版社,2014:41-42.
[14]Brooks SP,Trueman RC,Dunnett SB.Assessment of motor coordination and balance in mice using the rotarod,elevated bridge,and footprint tests[J].Curr Protoc Mouse Biol,2012,2(1):37-53.
[15]Vorhees CV,Williams MT.Morris water maze:procedures for assessing spatial and related forms of learning and memory[J].Nat Protoc,2006,1(2):848-858.
[16]Stradecki-Cohan HM,Youbi M,Cohan CH,et al.Physical exercise improves cognitive outcomes in 2 models of transient cerebral ischemia[J].Stroke,2017,48(8):2306.
[17]Wang Z,Guo Y,Myers KG,et al.Recruitment of the prefrontal cortex and cerebellum in Parkinsonian rats following skilled aerobic exercise[J].Neurobiol Dis,2015,77:71-87.
[18]Li F,Shi W,Zhao EY,et al.Enhanced apoptosis from early physical exercise rehabilitation following ischemic stroke[J].JNeurosci Res,2017,95(4):1017-1024.
[19]Ishrat T,Pillai B,Soliman S,et al.Low-dose candesartan enhances molecular mediators of neuroplasticity and subsequent functional recovery after ischemic stroke in rats[J].Mol Neurobiol,2015,51(3):1-12.
[20]Abe T,Kunz A,Shimamura M,et al.The neuroprotective effect of prostaglandin E2 EP1 receptor inhibition has a wide therapeutic window,is sustained in time and is not sexually dimorphic[J].J Cereb Blood Flow Metab,2009,29(1):66.
[21]Fluri F,Malzahn U,Homola GA,et al.Stimulation of the mesencephalic locomotor region for gait recovery after stroke[J].Ann Neurol,2017,82(5):828-840.
[22]Tesla R,Wolf HP,Xu P,et al.Neuroprotective efficacy of aminopropyl carbazoles in a mouse model of amyotrophic lateral sclerosis[J].P Natl Acad Sci USA,2012,109(42):17016.
[23]Wang Y,Bontempi B,Hong SM,et al.A comprehensive analysis of gait impairment after experimental stroke and the therapeutic effect of environmental enrichment in rats[J].J Cereb Blood Flow Metab,2008,28(12):1936-1950.
[24]侯志涛,孙忠人,刘松涛,等.电针对缺血性学习记忆障碍大鼠氧自由基及凋亡相关蛋白表达的影响[J].针刺研究,2015,40(6):431-438.
[25]Xia WG,Zhang CJ,Zhang X,et al.Effects of“nourishing liver and kidney”acupuncture therapy on expression of brain derived neurotrophic factor and synaptophysin after cerebral ischemia reperfusion in rats[J].华中科技大学学报:医学英德文版.2017(2):271-278.
[26]Tian T,Sun Y,Wu H,et al.Acupuncture promotes mTOR-independent autophagic clearance of aggregation-prone proteins in mouse brain[J].Sci Rep,2016,6:19714.
[2]董秋菊,杨志新.醒脑开窍针刺法治疗中风的系统评价[J].中国针灸,2013,33(5):475-480.
[3]岳增辉,李良,常小荣,等.电针与手针治疗脑卒中痉挛性瘫痪效应差异研究[J].中国针灸,2012,32(7):582-586.
[4]张通.中国脑卒中康复治疗指南(2011完全版)[J].中国康复理论与实践,2012,18(4):301-318.
[5]Li F,Shi W,Zhao EY,et al.Enhanced apoptosis from early physical exercise rehabilitation following ischemic stroke[J].JNeurosci Res,2017,95(4):1017-1024.
[6]葛亚博,冯晓东.电针疗法联合康复运动训练治疗脑卒中肢体痉挛大鼠的机制[J].中国老年学杂志,2017,37(15):3643-3645.
[7]杨慎峭,冯丽娟,毛雪莲,等.电针结合康复训练对脑卒中肢体痉挛大鼠突触结合蛋白Ⅰ表达的影响研究[J].时珍国医国药,2013,24(9):2300-2303.
[8]Tamura A,Graham DI,Mcculloch J,et al.Focal cerebral ischaemia in the rat:1.Description of technique and early neuropathological consequences following middle cerebral artery occlusion[J].J Cerebr Blood F Met,1981,1(1):53.
[9]Bederson JB,Pitts LH,Tsuji M,et al.Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J].Stroke,1986,17(3):472.
[10]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:254.
[11]曹勇.针刺对HIBD模型大鼠远期行为学及Ca2+浓度、LDH活性的影响[D].广州:广州中医药大学,2014.
[12]刘龙琳.针刺对全脑缺氧缺血大鼠海马区自噬相关蛋白的影响研究[D].广州:广州中医药大学,2014.
[13]袁青.靳三针法[M].北京:人民卫生出版社,2014:41-42.
[14]Brooks SP,Trueman RC,Dunnett SB.Assessment of motor coordination and balance in mice using the rotarod,elevated bridge,and footprint tests[J].Curr Protoc Mouse Biol,2012,2(1):37-53.
[15]Vorhees CV,Williams MT.Morris water maze:procedures for assessing spatial and related forms of learning and memory[J].Nat Protoc,2006,1(2):848-858.
[16]Stradecki-Cohan HM,Youbi M,Cohan CH,et al.Physical exercise improves cognitive outcomes in 2 models of transient cerebral ischemia[J].Stroke,2017,48(8):2306.
[17]Wang Z,Guo Y,Myers KG,et al.Recruitment of the prefrontal cortex and cerebellum in Parkinsonian rats following skilled aerobic exercise[J].Neurobiol Dis,2015,77:71-87.
[18]Li F,Shi W,Zhao EY,et al.Enhanced apoptosis from early physical exercise rehabilitation following ischemic stroke[J].JNeurosci Res,2017,95(4):1017-1024.
[19]Ishrat T,Pillai B,Soliman S,et al.Low-dose candesartan enhances molecular mediators of neuroplasticity and subsequent functional recovery after ischemic stroke in rats[J].Mol Neurobiol,2015,51(3):1-12.
[20]Abe T,Kunz A,Shimamura M,et al.The neuroprotective effect of prostaglandin E2 EP1 receptor inhibition has a wide therapeutic window,is sustained in time and is not sexually dimorphic[J].J Cereb Blood Flow Metab,2009,29(1):66.
[21]Fluri F,Malzahn U,Homola GA,et al.Stimulation of the mesencephalic locomotor region for gait recovery after stroke[J].Ann Neurol,2017,82(5):828-840.
[22]Tesla R,Wolf HP,Xu P,et al.Neuroprotective efficacy of aminopropyl carbazoles in a mouse model of amyotrophic lateral sclerosis[J].P Natl Acad Sci USA,2012,109(42):17016.
[23]Wang Y,Bontempi B,Hong SM,et al.A comprehensive analysis of gait impairment after experimental stroke and the therapeutic effect of environmental enrichment in rats[J].J Cereb Blood Flow Metab,2008,28(12):1936-1950.
[24]侯志涛,孙忠人,刘松涛,等.电针对缺血性学习记忆障碍大鼠氧自由基及凋亡相关蛋白表达的影响[J].针刺研究,2015,40(6):431-438.
[25]Xia WG,Zhang CJ,Zhang X,et al.Effects of“nourishing liver and kidney”acupuncture therapy on expression of brain derived neurotrophic factor and synaptophysin after cerebral ischemia reperfusion in rats[J].华中科技大学学报:医学英德文版.2017(2):271-278.
[26]Tian T,Sun Y,Wu H,et al.Acupuncture promotes mTOR-independent autophagic clearance of aggregation-prone proteins in mouse brain[J].Sci Rep,2016,6:19714.