重复经颅磁刺激对脑梗死大鼠恐惧记忆和海马区突触素表达的影响
发布时间：2019-04-22 11:51
【摘要】：目的:脑梗死(Cerebral infarction,CI)是指各种因素所致脑神经组织血液循环受阻,引起相应脑组织区域发生缺血缺氧性坏死,进而出现运动功能障碍、认知障碍、失语、共济失调等相应支配区域的功能障碍表现。药物和及时的手术治疗可以使患者的死亡率降低,挽救患者的生命,但对于已经损伤的脑组织引起的后遗症如认知功能障碍、运动功能障碍、感觉功能障碍、言语功能障碍等往往很难得到根治,严重影响患者日常生活活动能力,导致患者的生存质量下降。重复经颅磁刺激(repetitive Transcranial Magnetic Stimulation,rTMS)作为新的神经系统非药物治疗技术,具有无痛、非创伤、刺激效能稳定等物理特性,可达到缓解多种中枢神经系统疾病,促进患者的功能的恢复的作用,目前已广泛应用于治疗卒中后康复、神经性疼痛、外周神经疾病、癫痫、脊髓损伤等疾病,并取得了较好的成效。本试验分为两部分,旨在研究经重复颅磁刺激对脑梗死后大鼠记忆功能及海马区突触素表达的的影响,为rTMS进一步用于促进脑梗死患者的功能恢复提供理论指导依据。方法:成年雄性SD大鼠75只,体重约200~250g,参照线栓法对大鼠制作右侧大脑中动脉闭塞的脑缺血再灌注模型。造模成功的大鼠随机分为模型组及40%(0.76T)rTMS磁刺激组、60%(1.14T)rTMS磁刺激组、80%(1.52T)最大强度rTMS刺激组,每组15只,另外另设假手术组15只。各刺激组大鼠于建模成功后第二天按照设定参数分别接受rTMS治疗,连续5天。假手术组和模型组则不给予rTMS治疗。治疗结束后,各组随机抽取10只大鼠接受相同足部电击,在24h后记录大鼠在5分钟内的僵直持续时间;各组另外5只于治疗后1 d、7 d、14d、21 d、28 d通过免疫组化检测大鼠梗死侧海马区突触素(Synaptic bundle,SYN)的表达,最后对产生的数据进行统计学分析。结果:1.在恐惧记忆实验中:假手术组大鼠在接受足部电击24h后的5min内僵直持续时间均明显高于刺激组和模型组(p0.01);而各rTMS刺激组大鼠僵直持续时间均高于模型组;在刺激组中,60%(1.14T)磁刺激组的僵直持续时间最长。2.荧光显微镜观察:大鼠海马区的SYN免疫阳性物在假手术组数量最多,并在各个时间段分布均匀,且密度变化与作用时间无明显关联(p0.05)。模型组和刺激组SYN免疫阳性物计数短期内无明显差别,但随着rTMS作用于大脑中枢神经系统的时间延长,两组大鼠海马CA1区SYN免疫阳性物均出现明显增长,且刺激组SYN免疫阳性物的增幅明显高于模型组;在rTMS刺激组中,60%(1.14T)磁刺激组SYN阳性细胞计数相对最多,增幅最大(p0.05)。结论:1.重复经颅磁刺激(rTMS)对脑梗死大鼠记忆功能的恢复有一定的促进作用;2.rTMS对脑梗死大鼠梗死侧海马区突触素的表达有一定的促进作用;3.中等强度的rTMS对脑梗死大鼠记忆功能的恢复程度最高、海马区突触素阳性细胞数递增速度最快。
[Abstract]:Objective: cerebral infarction (Cerebral infarction,CI) refers to cerebral nerve tissue blood circulation obstruction caused by various factors, resulting in ischemic and hypoxic necrosis in the corresponding brain tissue area, and then motor dysfunction, cognitive impairment, aphasia, and so on. The dysfunctional manifestations of the corresponding dominant regions, such as ataxia. Medication and timely surgical treatment can reduce the mortality rate and save the life of the patient, but for the sequelae caused by brain damage, such as cognitive dysfunction, motor dysfunction, sensory dysfunction, Speech dysfunction is often difficult to cure, seriously affecting patients' ability of daily life, resulting in a decline in the quality of life of patients. Repetitive transcranial magnetic stimulation (repetitive Transcranial Magnetic Stimulation,rTMS), as a new non-drug therapy for nervous system, has the physical properties of painless, non-invasive and stable stimulation efficiency, which can alleviate many kinds of central nervous system diseases. At present, it has been widely used in the treatment of post-stroke rehabilitation, neurological pain, peripheral nerve diseases, epilepsy, spinal cord injury and other diseases, and has achieved good results. This experiment is divided into two parts. The purpose of this study is to study the effect of repeated cranial magnetic stimulation on memory function and synaptophysin expression in hippocampus of rats with cerebral infarction, and to provide theoretical guidance for the further use of rTMS in promoting the functional recovery of patients with cerebral infarction. Methods: 75 adult male SD rats, weighing about 200? 250 g, were used to establish the right middle cerebral artery occlusion (MCAO) cerebral ischemia reperfusion model. The successful rats were randomly divided into model group, 40% (0.76T) rTMS magnetic stimulation group, 60% (1.14T) rTMS magnetic stimulation group, 80% (1.52T) maximum intensity rTMS stimulation group (15 rats in each group), and 15 rats in sham operation group. The rats in each stimulation group were treated with rTMS according to the set parameters on the second day after the establishment of the model, and the rats in each stimulation group were treated for 5 consecutive days. Sham operation group and model group were not treated with rTMS. After treatment, 10 rats in each group were randomly selected to receive the same foot shock, and the duration of stiffness within 5 minutes was recorded 24 hours later. The expression of synaptophysin (Synaptic bundle,SYN) in the ipsilateral hippocampus of rats was detected by immunohistochemistry on day 1, day 7, day 14, day 21 and day 28 after treatment, and the data were analyzed statistically. Results: 1. In fear memory experiment, the duration of 5min stiffness in sham operation group was significantly longer than that in stimulation group and model group (p0.01) 24 hours after foot shock, while the duration of stiffness in each rTMS stimulation group was higher than that in model group, while the duration of stiffness in sham operation group was significantly higher than that in model group (p0.01). In the stimulation group, 60% (1.14T) magnetic stimulation group had the longest duration of rigidity. 2. Fluorescence microscope observation: the number of SYN immunoreactive substances in the hippocampus of rats was the most in the sham operation group, and the distribution was uniform in each period of time, and there was no significant correlation between the change of density and the time of action (p0.05). There was no significant difference in the number of SYN immunopositive substances between the model group and the stimulation group in a short time, but with the prolongation of the time of rTMS acting on the central nervous system of the brain, the SYN immunoreactivity in the hippocampal CA1 region of the two groups increased significantly. The increase of SYN immunoreactivity in stimulation group was significantly higher than that in model group. In the rTMS stimulation group, 60% (1.14T) magnetic stimulation group had the highest number of SYN positive cells and the largest increase (p0.05). Conclusions: 1. Repeated transcranial magnetic stimulation (rTMS) could promote the recovery of memory function in rats with cerebral infarction, 2.rTMS could promote the expression of synaptophysin in the hippocampus of rats with cerebral infarction, and (3) it could promote the expression of synaptophysin in the hippocampus of rats with cerebral infarction. The recovery of memory function by moderate intensity rTMS was the highest in cerebral infarction rats, and the increase rate of synaptophysin positive cells in hippocampus was the fastest.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.33
本文编号：2462811
[Abstract]:Objective: cerebral infarction (Cerebral infarction,CI) refers to cerebral nerve tissue blood circulation obstruction caused by various factors, resulting in ischemic and hypoxic necrosis in the corresponding brain tissue area, and then motor dysfunction, cognitive impairment, aphasia, and so on. The dysfunctional manifestations of the corresponding dominant regions, such as ataxia. Medication and timely surgical treatment can reduce the mortality rate and save the life of the patient, but for the sequelae caused by brain damage, such as cognitive dysfunction, motor dysfunction, sensory dysfunction, Speech dysfunction is often difficult to cure, seriously affecting patients' ability of daily life, resulting in a decline in the quality of life of patients. Repetitive transcranial magnetic stimulation (repetitive Transcranial Magnetic Stimulation,rTMS), as a new non-drug therapy for nervous system, has the physical properties of painless, non-invasive and stable stimulation efficiency, which can alleviate many kinds of central nervous system diseases. At present, it has been widely used in the treatment of post-stroke rehabilitation, neurological pain, peripheral nerve diseases, epilepsy, spinal cord injury and other diseases, and has achieved good results. This experiment is divided into two parts. The purpose of this study is to study the effect of repeated cranial magnetic stimulation on memory function and synaptophysin expression in hippocampus of rats with cerebral infarction, and to provide theoretical guidance for the further use of rTMS in promoting the functional recovery of patients with cerebral infarction. Methods: 75 adult male SD rats, weighing about 200? 250 g, were used to establish the right middle cerebral artery occlusion (MCAO) cerebral ischemia reperfusion model. The successful rats were randomly divided into model group, 40% (0.76T) rTMS magnetic stimulation group, 60% (1.14T) rTMS magnetic stimulation group, 80% (1.52T) maximum intensity rTMS stimulation group (15 rats in each group), and 15 rats in sham operation group. The rats in each stimulation group were treated with rTMS according to the set parameters on the second day after the establishment of the model, and the rats in each stimulation group were treated for 5 consecutive days. Sham operation group and model group were not treated with rTMS. After treatment, 10 rats in each group were randomly selected to receive the same foot shock, and the duration of stiffness within 5 minutes was recorded 24 hours later. The expression of synaptophysin (Synaptic bundle,SYN) in the ipsilateral hippocampus of rats was detected by immunohistochemistry on day 1, day 7, day 14, day 21 and day 28 after treatment, and the data were analyzed statistically. Results: 1. In fear memory experiment, the duration of 5min stiffness in sham operation group was significantly longer than that in stimulation group and model group (p0.01) 24 hours after foot shock, while the duration of stiffness in each rTMS stimulation group was higher than that in model group, while the duration of stiffness in sham operation group was significantly higher than that in model group (p0.01). In the stimulation group, 60% (1.14T) magnetic stimulation group had the longest duration of rigidity. 2. Fluorescence microscope observation: the number of SYN immunoreactive substances in the hippocampus of rats was the most in the sham operation group, and the distribution was uniform in each period of time, and there was no significant correlation between the change of density and the time of action (p0.05). There was no significant difference in the number of SYN immunopositive substances between the model group and the stimulation group in a short time, but with the prolongation of the time of rTMS acting on the central nervous system of the brain, the SYN immunoreactivity in the hippocampal CA1 region of the two groups increased significantly. The increase of SYN immunoreactivity in stimulation group was significantly higher than that in model group. In the rTMS stimulation group, 60% (1.14T) magnetic stimulation group had the highest number of SYN positive cells and the largest increase (p0.05). Conclusions: 1. Repeated transcranial magnetic stimulation (rTMS) could promote the recovery of memory function in rats with cerebral infarction, 2.rTMS could promote the expression of synaptophysin in the hippocampus of rats with cerebral infarction, and (3) it could promote the expression of synaptophysin in the hippocampus of rats with cerebral infarction. The recovery of memory function by moderate intensity rTMS was the highest in cerebral infarction rats, and the increase rate of synaptophysin positive cells in hippocampus was the fastest.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.33
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