血管内皮生长因子在动脉粥样硬化基础上缺血性脑卒中模型大鼠脑组织中的表达研究

发布时间：2018-06-16 08:11

本文选题：脑梗死 + 动脉粥样硬化动物模型　；参考：《山西医科大学》2012年硕士论文

【摘要】：目的探讨血管内皮生长因子基因及蛋白在动脉粥样硬化基础上缺血性脑卒中模型大鼠脑组织中的表达规律及意义。方法大鼠被随机分为空白对照组和模型组，后者制备动脉粥样硬化和缺血性卒中模型，然后再分为动脉粥样硬化模型假手术组（AS-sham）、动脉粥样硬化模型缺血性脑卒中持续缺血（AS-SI）0.5、6、12、24h组。观察主动脉根部及脑组织HE染色切片研究缺血性脑卒中和动脉粥样硬化模型的组织学上的基本病理改变，运用逆转录聚合酶链式反应技术（RT-PCR）检测血管内皮生长因子mRNA表达水平及其规律，免疫组织化学法被用于检测血管内皮生长因子蛋白的表达水平差异及其规律。计量资料结果以均数±标准差（x±s）表示，数据分析采用SPSS13.0软件。组间比较采用单因素方差分析，满足方差齐性条件，多重比较采用LSD-t法；不满足用Dunnett’sT3检验，差异以P0.05为标准。结果1.大体观：脑组织随着缺血时间的延长先出现水肿，后出现脑组织萎缩液化坏死；2.HE染色：随着缺血时间的延长神经元萎缩、坏死或者凋亡，组织出现液化坏死，，组织间坏死出现空白区；3.RT-PCR及免疫组化检测：各动脉粥样硬化模型组与对照组比较、各持续缺血组与假手术组比较血管内皮生长因子基因及蛋白表达均有明显升高（P0.05）,并随时间发生规律性变化，即0.5h迅速升高，其峰值在6h出现，12、24h后开始下降但仍较基线水平高。其中血管内皮生长因子基因及其蛋白产物在持续缺血6h组达到峰值。模型组可见双侧大脑皮质层、微血管周围脑组织及海马区细胞广泛分布有阳性表达颗粒，细胞水平可见胞浆内亦存在阳性表达颗粒。结论1.固态高脂饲料可成功复制稳定的动脉粥样硬化大鼠模型，在此基础上进一步结扎双侧颈总动脉可制备并发缺血性脑卒中的动脉粥样硬化大鼠模型，该模型属本实验课题首创。2.动脉粥样硬化基础上再发生缺血性脑卒中时两种疾病对VEGF表达的调节存在协同效应，VEGF表达的时相及水平较两种单纯疾病时表达的时相更早，水平更强烈。3.VEGF的表达与机体系统炎症的关系以及调节性机制需进一步研究，同时能提供在研究动脉粥样硬化基础上并发缺血性脑卒中的发生、发展、治疗及预后估计方面所需的理论模型。
[Abstract]:Objective to investigate the expression of vascular endothelial growth factor (VEGF) gene and protein in the brain of ischemic stroke rats on the basis of atherosclerosis. Methods Rats were randomly divided into blank control group and model group, the latter was used to establish atherosclerosis and ischemic stroke models, and then divided into the atherosclerosis model sham-operated group and the atherosclerotic model of ischemic stroke sustained ischemia. The histological changes of ischemic stroke and atherosclerosis model were studied by HE staining in the root of aorta and brain tissue. Reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the expression and regularity of vascular endothelial growth factor (VEGF) mRNA, and immunohistochemical method was used to detect the difference of expression level of vascular endothelial growth factor (VEGF) protein. The data were expressed as mean 卤standard deviation x 卤s, and SPSS 13.0 software was used to analyze the data. Single factor ANOVA was used to satisfy the condition of homogeneity of variance, and LSD-t method was used for multiple comparisons, and Dunnett-ST3 test was not satisfied. The difference was based on P05. Result 1. General observation: with the prolongation of ischemic time, brain tissue first appears edema, then brain tissue atrophy, liquefaction and necrosis. 2. He staining: with the prolongation of ischemic time, neuronal atrophy, necrosis or apoptosis, tissue liquefaction and necrosis, 3. RT-PCR and immunohistochemistry: the atherosclerosis model group was compared with the control group. Compared with sham operation group, the expression of VEGF gene and protein in each continuous ischemia group was significantly increased, and the expression of VEGF gene and protein increased regularly with time, that is, 0.5 h, the peak value began to decrease after 6 h and 1224 h, but still higher than baseline level. Vascular endothelial growth factor (VEGF) gene and its protein products reached the peak in 6 h ischemia group. In the model group, positive granules were found in bilateral cerebral cortex layer, brain tissue around microvessel and hippocampal area, and positive granules were also found in the cytoplasm of the model group. Conclusion 1. The stable rat model of atherosclerosis could be successfully established by solid high fat diet. On this basis, the rat model of atherosclerosis complicated with ischemic stroke could be made by further ligating bilateral common carotid artery. The model was the first one in this experiment. There is a synergistic effect on the regulation of VEGF expression between the two diseases on the basis of atherosclerosis. The time phase and level of VEGF expression are earlier than those of two simple diseases. The relationship between VEGF expression and systemic inflammation and its regulatory mechanism need to be further studied, which can provide the occurrence and development of ischemic stroke based on the study of atherosclerosis. Theoretical models needed for treatment and prognosis estimation.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363.2

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