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背景:血管内皮生长因子可加速新生血管形成,作为多功能细胞因子与其受体的相互作用在血管形成的过程中发挥关键作用。目的:检测血管内皮生长因子及其受体FLT-1及FLK-1m RNA 在急性局灶性脑缺血中的表达,并探讨其表达的时间与部位的相互关系。设计:随机对照实验。单位:吉林大学第一医院神经内科、吉林大学白求恩医学部病理教研室。材料:实验于2001-06/2002-04在吉林大学白求恩医学部病理实验室完成。选择成年SD 大鼠130只,雌雄各半,随机分为正常对照组10只,假手术组10只,脑缺血组110只。脑缺血组又随机分为0,1,2,3,6,24,48h,1,2,3,4周共11个时间点,每个时间点10只。方法:应用左侧颈总动脉结扎加缺氧诱导的方法,建立SD 大鼠永久大脑中动脉闭塞模型,假手术组不进行动脉结扎和缺氧处理,其余步骤同脑缺血组。正常对照组不做任何处理。检测血管内皮生长因子及其受体基因在局灶性脑缺血后不同时间点的表达应用原位杂交技术。同时观察局灶性脑缺血后血管形成情况。主要观察指标:①血管内皮生长因子及其受体基因在急性局灶性脑缺血后不同时间点的表达。②急性局灶性脑缺血后不同时间点血管形成情况。结果:130只大鼠全部进入结果分析。①不同时间点血管内皮生长因子m RNA 的表达:缺血3,6,24,48h 时缺血周边区高于缺血中心区犤(31.13±2.21)个/视野,(13.32±1.31)个/视野;(43.11±2.43)个/视野,(19.40±3.22)个/视野;(85.41±2.75)个/视野,(47.63±2.45)个/视野;(98.66±1.76)个/视野,(57.32±3.35)个/视野,(P <0.05)犦。48h 后血管内皮生长因子m RNA逐渐下降,至2周恢复到对照水平。②不同时间点血管内皮生长因子受体FLT-1,FLK-1m RNA 的表达:缺血3,6,24,48h 时缺血周边区高于缺血中心区,血管内皮生长因子受体FLT-1及FLK -1m RNA 表达在3周时达对照水平(P <0.05)。②血管形成平均数:48h,1周时缺血周边区高于缺血中心区犤(47.2±2.11)个/视野,(29.4±2.37) 个/视野;(199.2±3.45)个/视野,(76.6±4.62)个/视野,(P <0.05)犦。结论:急性局灶性脑缺血早期血管内皮生长因子m RNA 及血管内皮生长因子受体FLT-1及FLK-1m RNA 在神经细胞、胶质细胞、血管内皮细胞均有表达,在缺氧情况下,脑内血管内皮生长因子及受体的表达明显增强,表达具有时空特性。
BACKGROUND: Vascular endothelial growth factor (VEGF) accelerates neovascularization and plays a key role as an interaction of multifunctional cytokines with its receptors during angiogenesis. OBJECTIVE: To detect the expression of vascular endothelial growth factor and its receptors FLT-1 and FLK-1m RNA in acute focal cerebral ischemia, and to explore the relationship between the expression time and location. Design: Randomized controlled experiment. Unit: Department of Neurology, First Hospital of Jilin University, Department of Pathology, Bethune Medical College, Jilin University. Materials: The experiment was performed at Pathology Laboratory of Bethune Medical College, Jilin University from June 2001 to April 2002. 130 adult male Sprague-Dawley rats were randomly divided into normal control group (n = 10), sham operation group (n = 10) and cerebral ischemia group (n = 110). Cerebral ischemia group were randomly divided into 0, 1, 2, 3, 6, 24, 48h, 1, 2, 3, 4 weeks for a total of 11 time points, each time point 10. Methods: The model of middle cerebral artery occlusion in SD rats was established by ligation of the common carotid artery and hypoxia in the left. Artery ligation and hypoxia were performed in the sham operation group. The other steps were the same as those in the cerebral ischemia group. Normal control group without any treatment. Detection of vascular endothelial growth factor and its receptor gene expression at different time points after focal cerebral ischemia using in situ hybridization. At the same time, observe the situation of blood vessel formation after focal cerebral ischemia. MAIN OUTCOME MEASURES: ① The expression of vascular endothelial growth factor and its receptor genes at different time points after acute focal cerebral ischemia. ② angiogenesis at different time points after acute focal cerebral ischemia. Results: 130 rats all entered the result analysis. (1) The expression of vascular endothelial growth factor m RNA at different time points: The ischemic peripheral area at 3, 6, 24 and 48h after ischemia was significantly higher than that in ischemic center (31.13 ± 2.21) / field, (13.32 ± 1.31) (43.11 ± 2.43) / field, (19.40 ± 3.22) / field of vision, (85.41 ± 2.75) / field of vision, (47.63 ± 2.45) / field of vision, (98.66 ± 1.76) / field of vision, (57.32 ± 3.35) / vision, (P <0.05) 犦. 48h after vascular endothelial growth factor mRNA gradually decreased to 2 weeks returned to the control level. ② The expression of FLT-1 and FLK-1mRNA at different time points: The ischemic peripheral area was higher than the ischemic center at 3, 6, 24 and 48h after ischemia, and the expression of FLT- 1 and FLK-1mRNA expression reached the control level at 3 weeks (P <0.05). (2) The average number of vessel formation at 48h was higher than that in the ischemic central area (47.2 ± 2.11) / field, (29.4 ± 2.37) / field, (199.2 ± 3.45) / field at 1 week 76.6 ± 4.62) per visual field, (P <0.05) 犦. CONCLUSION: The expression of vascular endothelial growth factor mRNA and vascular endothelial growth factor receptor FLT-1 and FLK-1mRNA in early stage of acute focal cerebral ischemia are expressed in nerve cells, glial cells and vascular endothelial cells. In the condition of hypoxia , The expression of vascular endothelial growth factor and its receptor in the brain was significantly enhanced, and its expression had the characteristics of space-time.