大鼠线栓法大脑中动脉闭塞模型改进的实验研究

发布时间：2018-03-09 10:45

本文选题：脑缺血再灌注损伤　切入点：大脑中动脉闭塞　出处：《苏州大学》2011年硕士论文　论文类型：学位论文

【摘要】：背景与目的缺血性脑血管病(Ischemic Cerebral Vesscular Disease,ICVD)是神经科常见病多发病,在ICVD的治疗中重建血流或增强缺血区的血流供应是缺血脑组织修复损伤的必需条件,但同时带来的再灌注损伤也是目前最受关注的问题。线栓法大鼠大脑中动脉闭塞(middle cerebral artery occlusion, MCAO)模型能最大程度模拟人类缺血性卒中发生,被广泛用于实验性脑缺血再灌注(Ischemia-reperfusion ,IR)损伤的研究。国内外大量实验证实,MK-801(dizocilpine)在实验性脑缺血模型中具有重要的神经保护作用。本实验用周龄和性别匹配的SD大鼠成功建立了MCAO动物模型,对原有线栓法大脑中动脉闭塞模型进行了改进,并用MK-801预处理进一步验证动物模型,为研究脑缺血再灌注损伤的发病机理和研制治疗药物打下基础。方法 50只雄性SD大鼠随机分为3组,IR组、p-MCAO(permanent MCAO)组及假手术组(10只)。IR组(n=22)分为2个亚组,IR治疗组(12只)及IR模型组(10只)。p-MCAO组(n=18)也分为2个亚组,p-MCAO治疗组(10只)及p-MCAO模型组(8只)。两治疗组将药物MK-801(0.5mg/Kg,溶于二甲基亚砜:0.9%生理盐水,体积比为1:39的混合溶液中)于脑缺血15分钟前经颈外动脉给药。假手术组、两模型组大鼠仅给予等容量溶剂。手术组以线栓法制备MCAO模型,假手术组接受相同手术流程,但不插入线栓。手术后分别对各组大鼠做神经功能缺损评分,随后取各大鼠大脑冠状位切片行TTC染色以测量脑梗死及水肿体积,冰冻切片焦油紫染色观察大鼠大脑皮层区细胞损伤程度,酶联免疫吸附法测定脑中肿瘤坏死因子α(TNF-α)含量。结果 1.各手术组大鼠出现严重的神经功能障碍,假手术组大鼠则无任何临床症状。且神经行为学评分显示IR治疗组大鼠神经功能明显优于IR模型组。 2.切片TTC、焦油紫染色各手术组大鼠脑组织见明显脑梗死灶,假手术组大鼠无此改变。IR治疗组脑梗塞体积较IR模型组明显减小。 3.对大鼠脑组织生化指标检测发现,手术组大鼠的TNF-α的表达明显增加。与IR模型组相比,IR治疗组大鼠TNF-α的含量有所降低(P0.01)。结论 1.本实验建立的大鼠模型在临床表现、病理变化等方面符合MCAO模型的表现,模型稳定、可靠,是研究脑缺血再灌注损伤的理想动物模型。 2. MK-801能改善脑缺血再灌注损伤大鼠的神经功能障碍,减轻缺血区的脑梗死,并减少炎症因子TNF-α的产生,具有明显的改善脑缺血再灌注损伤的作用。
[Abstract]:Background and purpose. Ischemic Cerebral Vesscular disease (ICVD) is a common disease in neurology. In the treatment of ICVD, it is necessary to reconstruct the blood flow or increase the blood flow supply in the ischemic area. The middle cerebral artery occlusion (MCAO) model of middle cerebral artery occlusion (MCAO) can best mimic the occurrence of ischemic stroke in humans. MK-801 dizocilpine has been widely used in the study of experimental cerebral ischemia-reperfusion injury. A large number of experiments at home and abroad have confirmed that MK-801 dizocilpine plays an important role in the neuroprotection of experimental cerebral ischemia model. In this experiment, MCAO animal model was successfully established by week old and sex matched SD rats, the original middle cerebral artery occlusion model was improved, and the animal model was further verified by MK-801 pretreatment. To study the pathogenesis of cerebral ischemia-reperfusion injury and the development of treatment drugs. Method. 50 male Sprague-Dawley rats were randomly divided into 3 groups: ir group (n = 10) and sham-operated group (n = 10). They were divided into 2 subgroups (n = 12) and IR group (n = 10) and IR model group (n = 10). They were also divided into 2 subgroups (n = 10) and p-MCAO treatment group (n = 10) and p-MCAO model group (n = 10) and p-MCAO model group (n = 10). In the two treatment groups, the drug MK-801 + 0.5 mg / kg was dissolved in 0.9% normal saline of dimethyl sulfoxide (DMSO). The rats in sham-operated group and two model groups were given the same volume solvent only. The MCAO model was made by thread embolization in the operation group, and the sham-operation group received the same operation procedure. However, no thread embolus was inserted. After the operation, the neurological impairment scores were scored in each group of rats respectively. The coronal sections of the brain of each group were then taken for TTC staining to measure the volume of cerebral infarction and edema. The degree of cell injury in rat cerebral cortex was observed by tar violet staining and the content of TNF- 伪 in brain was determined by enzyme linked immunosorbent assay (Elisa). Results. 1. There were severe neurological dysfunction in each operation group, but no clinical symptoms in sham-operation group, and neurobehavioral score showed that the nerve function of IR group was better than that of IR model group. 2. There were obvious cerebral infarction in brain tissue of rats in each operation group stained with TTCand tar violet, but no change was found in sham-operated group. The infarct volume in IR treatment group was significantly smaller than that in IR model group. 3. It was found that the expression of TNF- 伪 in the operation group was significantly higher than that in the IR model group, and the content of TNF- 伪 in the IR treatment group was lower than that in the IR model group. Conclusion. 1. The rat model established in this study accords with the MCAO model in clinical manifestations and pathological changes. The model is stable and reliable. It is an ideal animal model for the study of cerebral ischemia-reperfusion injury. 2. MK-801 can improve the neurological dysfunction of rats with cerebral ischemia reperfusion injury, reduce the cerebral infarction in the ischemic area, and reduce the production of inflammatory factor TNF- 伪, which can obviously improve the cerebral ischemia-reperfusion injury.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R743;R-332

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