【摘要】：目的:腦梗死(Cerebral infarction,CI)是指各種因素所致腦神經(jīng)組織血液循環(huán)受阻,引起相應腦組織區(qū)域發(fā)生缺血缺氧性壞死,進而出現(xiàn)運動功能障礙、認知障礙、失語、共濟失調(diào)等相應支配區(qū)域的功能障礙表現(xiàn)。藥物和及時的手術治療可以使患者的死亡率降低,挽救患者的生命,但對于已經(jīng)損傷的腦組織引起的后遺癥如認知功能障礙、運動功能障礙、感覺功能障礙、言語功能障礙等往往很難得到根治,嚴重影響患者日常生活活動能力,導致患者的生存質(zhì)量下降。重復經(jīng)顱磁刺激(repetitive Transcranial Magnetic Stimulation,rTMS)作為新的神經(jīng)系統(tǒng)非藥物治療技術,具有無痛、非創(chuàng)傷、刺激效能穩(wěn)定等物理特性,可達到緩解多種中樞神經(jīng)系統(tǒng)疾病,促進患者的功能的恢復的作用,目前已廣泛應用于治療卒中后康復、神經(jīng)性疼痛、外周神經(jīng)疾病、癲癇、脊髓損傷等疾病,并取得了較好的成效。本試驗分為兩部分,旨在研究經(jīng)重復顱磁刺激對腦梗死后大鼠記憶功能及海馬區(qū)突觸素表達的的影響,為rTMS進一步用于促進腦梗死患者的功能恢復提供理論指導依據(jù)。方法:成年雄性SD大鼠75只,體重約200~250g,參照線栓法對大鼠制作右側(cè)大腦中動脈閉塞的腦缺血再灌注模型。造模成功的大鼠隨機分為模型組及40%(0.76T)rTMS磁刺激組、60%(1.14T)rTMS磁刺激組、80%(1.52T)最大強度rTMS刺激組,每組15只,另外另設假手術組15只。各刺激組大鼠于建模成功后第二天按照設定參數(shù)分別接受rTMS治療,連續(xù)5天。假手術組和模型組則不給予rTMS治療。治療結(jié)束后,各組隨機抽取10只大鼠接受相同足部電擊,在24h后記錄大鼠在5分鐘內(nèi)的僵直持續(xù)時間;各組另外5只于治療后1 d、7 d、14d、21 d、28 d通過免疫組化檢測大鼠梗死側(cè)海馬區(qū)突觸素(Synaptic bundle,SYN)的表達,最后對產(chǎn)生的數(shù)據(jù)進行統(tǒng)計學分析。結(jié)果:1.在恐懼記憶實驗中:假手術組大鼠在接受足部電擊24h后的5min內(nèi)僵直持續(xù)時間均明顯高于刺激組和模型組(p0.01);而各rTMS刺激組大鼠僵直持續(xù)時間均高于模型組;在刺激組中,60%(1.14T)磁刺激組的僵直持續(xù)時間最長。2.熒光顯微鏡觀察:大鼠海馬區(qū)的SYN免疫陽性物在假手術組數(shù)量最多,并在各個時間段分布均勻,且密度變化與作用時間無明顯關聯(lián)(p0.05)。模型組和刺激組SYN免疫陽性物計數(shù)短期內(nèi)無明顯差別,但隨著rTMS作用于大腦中樞神經(jīng)系統(tǒng)的時間延長,兩組大鼠海馬CA1區(qū)SYN免疫陽性物均出現(xiàn)明顯增長,且刺激組SYN免疫陽性物的增幅明顯高于模型組;在rTMS刺激組中,60%(1.14T)磁刺激組SYN陽性細胞計數(shù)相對最多,增幅最大(p0.05)。結(jié)論:1.重復經(jīng)顱磁刺激(rTMS)對腦梗死大鼠記憶功能的恢復有一定的促進作用;2.rTMS對腦梗死大鼠梗死側(cè)海馬區(qū)突觸素的表達有一定的促進作用;3.中等強度的rTMS對腦梗死大鼠記憶功能的恢復程度最高、海馬區(qū)突觸素陽性細胞數(shù)遞增速度最快。
[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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R743.33

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