【作者】 杨玮春； 史鹤； 胡敏予；

【Author】 Weichun Yang;He Shi;Minyu Hu;

【机构】 中南大学公共卫生学院营养与食品卫生学系；

【摘要】 目的通过检测缺血情况下大鼠血清及脑S100B蛋白的水平,探讨缺血对血脑屏障的影响,为后续研究缺血对大脑的损伤提供理论依据。方法将18只雌雄各半成年SD大鼠,体重(200 50)g,随机分为3组,各组大鼠以1%戊巴比妥钠(45mg/kg·bw-1)腹腔注射麻醉,模型组行双侧颈总动脉结扎术,假手术组分离出双侧颈总动脉后不予以结扎,空白组不进行血管分离手术。3h后模型组及假手术组从股静脉注入辣根过氧化物酶(HRP),空白组则注入等量生理盐水,循环30min后采集血液及脑组织,检测大鼠血清及脑S100B蛋白含量,HE染色观察海马形态学变化,应用辣根过氧化物酶-二氨基联苯胺(HRP-DAB)显色法检测大鼠血脑屏障的通透性。结果模型组大鼠血清S100B蛋白水平(24.18±4.51ng/L)高于空白组(10.97±2.47ng/L)及假手术组(15.08±3.14ng/L)(F=22.579,P<0.05),脑S100B蛋白水平(566.41±94.69×10-5ng/mg)高于空白组(355.27±173.62×10-5ng/mg)及假手术组(375.95±121.48×10-5ng/mg)(F=4.527,P<0.05)。高倍镜下空白组海马锥体细胞排列整齐,假手术组细胞排列稍紊乱、层次结构欠清,模型组锥体细胞减少,排列紊乱,缺血损伤程度最严重。高倍镜下各组均难以分辨出HRP-DAB反应物。结论缺血造成大鼠血清及脑S100B蛋白水平增高,海马结构紊乱,细胞间的连接疏松,提示缺血可引起血脑屏障的损伤。本实验中运用HRP-DAB显色法评价血脑屏障通透性时,镜下难以辨别出HRP-DAB反应产物。更多还原

【Abstract】 Objective:To investigate the effects of ischemia on blood-brain barrier, by detecting the levels of serum and brain S100 B protein of rats, for subsequent ischemic brain injury study provides a theoretical basis.Methods: The 18 male and female adult Sprague Dawley(SD) rats weighing(200 50) g, were randomly divided into three groups(n=6), each group of rats with 1% sodium pentobarbital(45mg/kg·bw-1)anesthetized by intraperitoneal injection. In the model group, bilateral common carotid arteries were ligated, the sham-operation group bilateral carotid arteries not to be ligated after separation, the control group without separation surgery, 3h after the model group and the sham-operation group are injected into horseradish peroxidase(HRP) via femoral vein, the control group is injected into equal amount of saline, 30min after blood and brain tissue were collected, then for S100 B protein detection, HE staining for the hippocampal morphology changes, the application of horseradish peroxidase-diaminobenzidine(HRP-DAB) colorimetric method to detect blood-brain barrier permeability. Results: The level of S100 B protein in serum(24.18±4.51ng/L) of model group is higher than the control group(10.97±2.47ng/L) and sham-operation group(15.08±3.14ng/L)(F=22.579,P<0.05), the level of S100 B protein in brain(566.41±94.69×10-5ng/mg) of model group is higher than the control group(355.27±173.62×10-5ng/mg) and sham-operation group(375.95±121.48×10-5ng/mg)(F=4.527,P<0.05). Observed at high magnification, hippocampal pyramidal cells of the control group arranged in neat rows, the sham-operation group showed slightly disordered cell arrangement and less clear hierarchy, pyramidal cells in the model group reduced and disorganized, the most severe ischemic injury. At high magnification in each group are difficult to distinguish the HRP-DAB reaction. Conclusion:Ischemia can increase the levels of S100 B protein in theserum and brain of rat,lead to the disorder of hippocampal structure, also loose connections between cells, suggesting ischemia can cause blood-brain barrier damage. In this study, the application of HRP-DAB chromogenic method to evaluate the blood-brain barrier permeability is difficult to identify HRP-DAB reaction product in the microscope.更多还原