【作者】 李健；

【导师】 金荣疆；

【作者基本信息】 成都中医药大学 ， 康复医学与理疗学， 2018， 硕士

【摘要】 目的:本课题以大鼠局灶性脑缺血再灌注模型为研究对象,以水中运动疗法为干预方法,以随意运动障碍、神经功能评分、肌张力检测、组织学检查、γ-氨基丁酸(GABA)及谷氨酸(Glu)的含量为评价标准,观察水中运动疗法对脑组织抑制性神经递质以及兴奋性神经递质表达的影响,探索水中运动疗法治疗缺血性脑卒中后肢体偏瘫痉挛的可能作用机制,为水中运动疗法的临床应用提供理论基础,为水中运动疗法进一步研究提供参考。方法:将60只240-270 g SD大鼠,雌雄各半,按照MCAO模型大鼠:假手术组:空白组动物例数比例为4:1:1的比例随机分为三组,假手术组与空白组动物各10只。MCAO模型组大鼠经造模并经检验模型成功入组后,随机分为两组:模型组与治疗组。假手术组仅分离右侧颈总、颈外动脉和颈内动脉;空白组不做任何处理。建模成功后第4d,治疗组给予水中运动疗法治疗,每天30min,治疗6d;空白组、假手术组和模型组自由活动,不予任何治疗。各组大鼠在治疗前及治疗后分别进行随意运动障碍评分、神经功能评分和肌张力评定。术后10d处死大鼠,在超净冰面台上取出右侧大脑组织。在光镜下观察各组右侧大脑皮质的病理形态变化。以ELISA测定各组标本中GABA和Glu的表达情况。结果:1、神经功能评分和随意运动功能评分结果:治疗前,假手术组和空白组相比,差异无统计学意义(P>0.05);造模大鼠和空白组相比,差异有统计学意义(P<0.05),模型组与治疗组间无统计学差异;治疗后,治疗组较治疗前神经功能与运动功能恢复(P<0.05),模型组有恢复趋势;与模型组比较,治疗组评分均显著降低((P<0.05);2、肌张力评定:采用运动幅度描记评定肌张力。治疗前后假手术组和空白组相比,差异没有统计学意义(P>0.05);治疗前,与空白组比较,模型动物肌张力均有增高,差异具有统计学意义(P<0.05),治疗组与模型组间无明显差异(P>0.05);治疗后,治疗组较治疗前痉挛水平降低(P<0.05),模型组有降低趋势;与模型组比较,治疗组的肌张力有所改善,差异有统计学意义(P<0.05)。3、病理HE切片示:(1)模型组:脑组织广泛变性,神经细胞坏死,细胞质水肿,炎细胞浸润;部分区域可见血细胞漏出,小胶质细胞增生。(2)治疗组:脑组织依然可见广泛的液化性坏死,胞浆肿胀,镜下小胶质细胞明显增多。(3)假手术组及空白组大鼠神经组织未见明显异常。4.ELISA结果显示:GABA的表达:假手术组和空白组相比,差异无统计学意义(P>0.05);与空白组比较,治疗组大鼠脑组织中GABA的表达增加(P<0.05),模型组略增加,无统计学意义(P>0.05);与模型组比较,治疗组大鼠脑组织中GABA的表达增加(P<0.05)。Glu的表达:假手术组和空白组相比,差异无统计学意义(P>0.05);与空白组比较,模型组大鼠脑组织中Glu的表达增加(P<0.05),治疗组大鼠增加幅度无统计学意义(P>0.05);与模型组比较,治疗组大鼠脑组织中Glu的表达低(P<0.05)。结论:1、水中运动疗法能缓解缺血性卒中大鼠肢体痉挛,明显改善偏瘫大鼠的运动功能。2、水中运动疗法可加快坏死脑组织的修复,促进卒中大鼠神经功能的恢复大鼠脑组织内小胶质细胞数目的增多与其神经保护作用可能是其作用机制之一。3、水中运动疗法下调中枢Glu表达水平,Glu兴奋作用减弱可能是水中运动疗法改善偏瘫肢体痉挛的作用机制之一。4、水中运动疗法调高中枢GABA表达水平,GABA抑制作用增强可能是水中运动疗法改善偏瘫肢体痉挛的作用机制之一。更多还原

【Abstract】 Objective:In this study,the model of focal cerebral ischemia reperfusion in rats was taken as the research object,and the method of exercise therapy in water was taken as an intervention method,with random movement obstacle,nerve function score,muscle tension,histological examination,gamma aminobutyric acid(GABA)and glutamic acid(Glu)content as evaluation criteria,to observe the exercise therapy in water.The effects of the inhibitory neurotransmitters and excitatory neurotransmitters on the expression of excitatory neurotransmitters in the treatment of cerebral hemiplegic spasm after ischemic stroke are explored to provide a theoretical basis for the clinical application of water exercise therapy and to provide a reference for the further study of water exercise therapy.Methods:A total of 60 240-270 g SD rats were divided into three groups according to the MCAO model group: sham-operation group and blank group.The ratio of animals to 4:1:1 was randomly divided into three groups.After grouping,there were 20 animals in the treatment group and the model group,and 10 animals in the sham-operation group and the blank group.MCAO model group rats were randomly divided into two groups: model group and treatment group.The middle cerebral artery occlusion(MACO)model was established in rats in the model group and treatment group.The sham-operation group only separated the right common carotid,external and internal carotid arteries,while the blank group did not do any treatment.After establishing model for 3days,the treatment group was treated with exercise therapy in water,30 min a day,lasting 6 days.The blank group,sham-operation group and model group were free to exercise,and no special treatment was given.The random movement obstacle score,nerve function score and muscle tone score were performed before and after treatment,respectively.At the end of the treatment,the rats were killed and the right brain tissue was removed on the super clean surface.The pathological changes of the right cerebral cortex in each group were observed under light microscope.The expression of GABA and Glu in each group was measured by ELISA.Results:1.Neurological score and voluntary motor function score results: before treatment,compared with control group and sham-operation group,the difference was not statistically significant(P > 0.05);compared with the blank group and model rats,the difference was statistically significant(P<0.05),the model group and the treatment groups had no statistically significant difference after treatment;compared with the model group,the treatment group scores were significantly lower((P<0.05);2.Muscle tone assessment: electrophysiography was used to assess muscle tone.Before treatment,compared with blank group and sham-operation group,the difference was not statistically significant(P>0.05);compared with the blank group,model group and treatment group increased muscle tone,the difference was statistically significant(P<0.05),but there was no significant difference between these groups(P>0.05);after modeling after treatment,compared with the model group,the muscle tone in treatment group improved,the difference was statistically significant(P<0.05)3.Pathological HE slices showed:(1)in model group,brain tissue was extensively denatured,the nucleus was mostly condensed,cytoplasmic edema and hemorrhage,inflammatory cells infiltration,blood cells leakage in some areas and microglia hyperplasia.(2)In the treatment group,there was still a wide range of liquefaction necrosis in the brain tissue,the nuclear condensation,the disappearance of nucleolus,the swelling of the cytoplasm,and the obvious increase of microglia under the microscope.(3)There was no obvious abnormality in the nerve tissue of the shamoperation group and the blank group.4.ELISA results showed :for the expression of GABA,compared with blank group and sham-operation group,the difference was not statistically significant(P>0.05);compared with the blank group,increased expression of GABA in brain tissue of rats in the model group and treatment group were statistically significant(P<0.05);compared with the model group,the expression of GABA in brain tissue of treatment group rats increased significantly(P<0.05).Glu expression: the difference between the sham-operation group and the blank group was not statistically significant(P>0.05).Compared with the blank group,the expression of Glu in the brain tissue of the model group increased(P<0.05),and the increased amplitude of the rats in the treatment group was not statistically significant(P>0.05).Compared with the model group,the expression of Glu in the brain tissue of the treatment group was low(P<0.05).Conclusion:1.Aquatic exercise therapy can relieve the limb spasm in ischemic stroke rats and improve the motor function of the hemiplegic rats.2.Aquatic exercise therapy can accelerate the repair of necrotic brain tissue,and promote the recovery of neural function in rats.The increase of microglia in the brain tissue of rats and its neuroprotective effect may be one of its mechanisms.3.Aquatic exercise therapy can reduce the expression level of Glu in the central nervous system.Glu may play an important role in the treatment of hemiplegic limb spasticity.4.The GABA expression level of high altitude pith in aquatic exercise therapy,and the increase of GABA inhibitory effect may be one of the mechanisms of water exercise therapy to improve hemiplegia limb spasm.更多还原