丰富环境对脑缺血再灌注损伤后脑血流和血管新生的影响
发布时间:2018-03-23 10:57
本文选题:丰富 切入点:环境 出处:《复旦大学》2014年硕士论文
【摘要】:目的:既往研究证实,缺血性脑梗塞后的血流恢复及血管再生对脑功能的改善有重要作用。本实验拟探讨丰富环境干预是否能改善脑卒中后血管新生和脑血流的改善。方法:将SD大鼠随机分为3组:假手术组、丰富环境组和标准环境组,并进行大脑中动脉梗塞手术,其中假手术组除不堵塞血管这一步骤,其余步骤与手术组相同。在缺血前和缺血1小时分别运用激光散斑技术检测各组大鼠大脑中动脉供血区皮层的相对血流速度,然后将手术后的大鼠分别放入相应的环境中饲养4周,4周后再次检测同一区域皮层相对血流速度。另有大鼠干预4周后经大鼠尾静脉注入异硫氰酸荧光素右旋糖苷(Fluorescein Isothiocyanate dextran, FITC-dextran),5分钟后断头取脑,固定后切片,通过激光共聚焦扫描显微镜检测缺血干预后的血管密度。同时,运用免疫印记(Western blot)技术检测缺血侧大脑皮层血管内皮生长因子(Vascular Endothelial Growth Factor, VEGF)的表达。结果:与缺血前比较,丰富环境组和标准环境组缺血侧皮层的相对血流速度在缺血1小时后明显下降(P0.01),假手术组无明显变化。干预4周后,与标准环境组相比,丰富环境组的缺血侧皮层血流速度明显增加(P0.05)。另外,丰富环境干预4周后的大鼠缺血侧皮层的血管密度明显大于标准环境组,VEGF蛋白表达也明显高于标准环境组。结论:丰富环境可以增加缺血后大鼠的缺血侧皮层血流,血流的增加可能与丰富环境促进了VEGF的表达从而增强血管再生有关。目的:血管新生是成人脑缺血后血管生成的最主要方式,也是现如今研究的主要方向;脑微血管内皮细胞是主要的脑血管组成成分,对脑缺血后的血管新生及其重要。本项研究将探讨丰富环境干预后大鼠的血清对氧糖剥夺后脑微血管内皮细胞的增殖作用。方法:丰富环境及标准环境干预同前,干预后大鼠心脏采血,离心后取上清,过滤后备用。培养脑微血管内皮细胞,实施氧糖剥夺实验以复制体外脑缺血模型。而后进行预实验,用10%、20%和40%浓度的不同干预血清分别培养氧糖剥夺后的脑微血管内皮细胞,用CCK-8测定其增殖,摸索最适浓度。最后,用最适浓度血清多次重复培养氧糖剥夺后的脑微血管内皮细胞,进一步探讨丰富环境干预后大鼠的血清对氧糖剥夺后的脑微血管内皮细胞的增殖作用。结果:10%浓度的干预血清培养氧糖剥夺后的脑微血管内皮细胞,其CCK-8检测的OD值明显大于20%和40%浓度的干预血清(P0.01);10%浓度丰富环境干预的大鼠血清培养氧糖剥夺后的脑微血管内皮细胞,其CCK-8检测的OD值明显大于标准环境干预的大鼠血清的CCK-8值(P0.01)。结论:10%浓度的干预血清对氧糖剥夺后的脑微血管内皮细胞的促增殖作用最明显;10%浓度丰富环境干预的大鼠血清对氧糖剥夺后的脑微血管内皮细胞的促增殖作用明显优于同浓度标准环境干预的大鼠血清。
[Abstract]:Objective: past studies have confirmed that, Blood flow recovery and vascular regeneration after ischemic cerebral infarction play an important role in the improvement of cerebral function. The purpose of this study was to explore whether rich environment intervention can improve angiogenesis and cerebral blood flow after stroke in SD rats. They were randomly divided into three groups: sham-operated group, Enrich the environment group and the standard environment group, and perform the middle cerebral artery infarction operation, the sham-operation group except the step of not blocking the blood vessel, The other steps were the same as those in the operation group. Laser speckle technique was used to detect the relative blood flow velocity in the cortex of the middle cerebral artery in each group before and after 1 hour of ischemia. Then the rats were fed in the corresponding environment for 4 weeks and 4 weeks later, the relative blood flow velocity of cortex in the same area was measured again. After 4 weeks of intervention, the dextrose of fluorescein isothiocyanate was injected into the caudal vein of the rats. Fluorescein Isothiocyanate dextran, FITC-dextranan, and brain were taken after 5 minutes. After fixation, the vascular density was detected by laser confocal scanning microscope. At the same time, The expression of vascular Endothelial Growth factor (VEGF) in ischemic cerebral cortex was detected by Western blot technique. The relative blood flow velocity of ischemic cortex in the rich environment group and the standard environment group decreased significantly after 1 hour of ischemia, but there was no significant change in the sham operation group. After 4 weeks of intervention, compared with the standard environment group, the relative blood flow velocity of the ischemic cortex decreased significantly in the rich environment group and the standard environment group. In the rich environment group, the blood flow velocity of ischemic cortex increased significantly (P 0.05). The vascular density of ischemic cortex of rats after 4 weeks of rich environment intervention was significantly higher than that of standard environment group, and the expression of VEGF protein was also significantly higher than that of standard environment group. Conclusion: rich environment can increase the blood flow of ischemic side cortex of rats after ischemia. The increase of blood flow may be related to the enhanced expression of VEGF and the enhancement of angiogenesis. Objective: angiogenesis is the most important way of angiogenesis after cerebral ischemia in adults, and it is also the main research direction nowadays. Cerebral microvascular endothelial cells are the main components of cerebral vessels. This study will explore the effect of serum enriched environment intervention on the proliferation of cerebral microvascular endothelial cells after oxygen glucose deprivation in rats. After intervention, the rat heart blood was collected, the supernatant was taken after centrifugation, and then filtered. The cerebral microvascular endothelial cells were cultured, and the oxygen glucose deprivation test was carried out to replicate the model of cerebral ischemia in vitro. Brain microvascular endothelial cells were cultured with different concentrations of 10% and 40% different concentrations of oxygen glucose deprivation, and their proliferation was determined by CCK-8, and the optimal concentration was found. Brain microvascular endothelial cells were cultured repeatedly with the optimal concentration of serum for several times after oxygen glucose deprivation. To further study the proliferation effect of serum of rats exposed to enriched environment on cerebral microvascular endothelial cells after oxygen glucose deprivation. Results the cultured cerebral microvascular endothelial cells were cultured with 10% concentration of 10% concentration of intervention serum. The OD value detected by CCK-8 was significantly higher than that of 20% and 40% of the intervention serum P0.01T10%, which enriched the concentration of cultured cerebral microvascular endothelial cells after oxygen glucose deprivation in rats. The OD value detected by CCK-8 was significantly higher than that of serum CCK-8 of rats treated with standard environmental intervention (P 0.01). Conclusion the concentration of 10% 10% of the intervention serum can promote the proliferation of cerebral microvascular endothelial cells after oxygen glucose deprivation, and the concentration of 10% can enrich the concentration of environmental intervention. The proliferative effect of rat serum on cerebral microvascular endothelial cells after oxygen glucose deprivation was significantly better than that of rats treated with the same concentration of standard environment.
【学位授予单位】:复旦大学
【学位级别】:硕士
【学位授予年份】:2014
【分类号】:R743
【共引文献】
相关期刊论文 前10条
1 于哲新;顾复生;潘德民;李禄金;陆宁;;北京社区居民健康体检人群冠心病危险因素分布评估[J];中国医药导刊;2014年07期
2 王艳飞;任越;刘海波;刘亚宁;冯翠娜;;普通造影导丝代替交换导丝经桡动脉途径PCI治疗对比分析[J];广西医学;2014年11期
3 许梦佳;顾卓雅;赵健元;王红艳;刘杰;陈瑛;;叶酸代谢通路关键酶基因多态性的肝癌易感性分析[J];复旦学报(自然科学版);2014年06期
4 徐晓东;孔晨晨;席正祺;;大数据云计算技术在全国机动车缉查布控系统中的应用[J];中国公共安全(学术版);2015年01期
5 王栋;倪光夏;;近10年针灸治疗脑卒中后肩手综合征研究进展[J];北京中医药;2015年04期
6 王志光;林丽珠;李新;王三虎;范先基;邢晓娟;杨子玉;;通补三升汤对Ⅲ期肺鳞癌同期放化疗患者无进展生存期的影响[J];广东医学;2015年09期
7 周军;吴e,
本文编号:1653161
本文链接:https://www.wllwen.com/yixuelunwen/shenjingyixue/1653161.html
最近更新
教材专著