【作者】 张思硕；

【导师】 郝斌；

【作者基本信息】 山西医科大学 ， 外科学， 2010， 硕士

【摘要】 缺氧诱导因子-1α(HIF-1α)、血管内皮细胞生长因子(VEGF)在血管生成表达中有重要意义,HIF-1α是缺氧状态下血管生成的核心调控因子,通过影响其他生长因子的表达参与血管生成的全过程。VEGF是血管生成初期关键性的生长因子,缺氧状态可以诱导VEGF大量表达。VEGF在血管生成中发挥重要作用,VEGF具有促血管内皮细胞分裂、增殖,增加血管的通透性,诱导内皮细胞Bcl-2基因表达,提高内皮细胞抗凋亡能力等作用。本实验通过检测缺氧诱导因子-1α(HIF-1α)和血管内皮细胞生成因子(VEGF)在兔颈总动脉内膜剥脱+颈前静脉自体静脉移植手术前后各期的表达以及移植静脉术后剥脱内膜段动脉管腔横截面积的变化,研究HIF-1α和VEGF在兔移植静脉再内皮化过程中的作用。目的探讨HIF-1α和VEGF的表达对大白兔移植静脉再内皮化的影响。1、HIF-1α和VEGF在兔颈总动脉内膜剥脱+颈前静脉自体静脉移植手术前后各期的表达关系。2、移植静脉术后剥脱内膜段动脉管腔横截面积的变化方法1、分组和制作动物模型6月龄成年雄性新西兰大白兔46只,随机分为两组,第一组10只,第二组36只(其中30只为试验必须动物,6只为备用)。均以普通饮食饲养一周后,采集第一组10只实验兔的血清分别采用酶联免疫吸附法(ELISA法)行HIF-1α和VEGF检测,然后取所有10只兔的左颈前静脉行HE染色做病理学检查,观察动脉血管腔面积。第二组36只大白兔均行左侧颈总动脉内膜剥脱+颈前静脉自体静脉移植术,制作再狭窄模型,术后以普通饮食饲养。于手术后7天、14天、30天三个时间点依次分别随机抽取10只实验兔采集血清用酶联免疫吸附法(ELISA法)行HIF-1α和VEGF检测,抽血后取该次试验10只兔剥脱内膜段动脉行HE染色做病理学实验,并观察动脉血管腔面积、内膜增生和平滑肌数量的动态变化。2、病理学资料图像分析采用病理学图像分析系统对标本：兔移植静脉HE染色、Mason染色片进行形态学分析。3、化学法分析分别于静脉移植手术前和手术后7天、14天、30天采血用ELISA法测量血清中HIF-1α和VEGF的水平。结果1、HIF-1α和VEGF各期的表达差别具有统计学意义(P<0.01)；HIF-1α和VEGF在兔自体静脉移植术后表达明显增加(P<0.01),其中在7天时最高,之后逐渐下降,在术后30天时仅稍高于正常。2、HE染色结果显示剥脱内膜段动脉血管腔面积(LA)总体呈先逐渐缩小,于术后14天达最低值,然后逐渐增大趋势。结论1、本次实验中成功地制作了兔颈总动脉内膜剥脱+自体静脉移植术后再狭窄模型。2、组织器官急性缺血缺氧期HIF-1α和VEGF表达迅速增高；随着时间延长,组织器官对缺血缺氧的适应,两因子浓度逐渐降低,本实验中观察到的HIF-1α和VEGF表达高峰出现在术后7天,且两指标在各期表达差别明显有统计学意义。3、移植静脉术后血管重塑的早期HIF-1α和VEGF的高表达可促进受损血管内皮细胞(VEC)结构和功能的重建,维持动脉管腔面积；随着时间推移,HIF-1α和VEGF的表达逐渐减少,移植静脉术后剥脱内膜段动脉管腔逐渐缩小,于术后14天达到最小,然后逐渐增大,且动脉管腔面积在各期差别明显有统计学意义。4、HIF-1α和VEGF表达高峰和移植静脉术后剥脱内膜段动脉管腔最小值分别出现在术后7天和14天,所以术后7-14天HIF-1α和VEGF表达的过早降低可能与移植静脉术后再狭窄有关。在各期HIF-1α和VEGF表水平均服从正态分布,经过Pearson相关分析存在强的正相关,相关系数r分别为0.901,0.832,0.779,0.863。更多还原

【Abstract】 Hypoxia-inducible factor-la (HIF-la), vascular endothelial growth factor (VEGF) expression in angiogenesis of significance, HIF-la is a hypoxia state the core of regulation of angiogenesis factors, by influencing other growth factors Expression of the entire process involved in angiogenesis. VEGF is a critical early stage of angiogenesis growth factor, hypoxia may induce substantial expression of VEGF.VEGF is a critical early stage of angiogenesis growth factor, hypoxia may induce substantial expression of VEGF. VEGF in the angiogenesis play an important role, VEGF has vascular endothelial cell division, proliferation, increase vascular permeability, induce endothelial cell Bcl-2 gene expression, enhance the ability of endothelial cells in anti-apoptotic effect.In this study, by detecting hypoxia-inducible factor-la (HIF-la) and vascular endothelial growth factor (VEGF) in rabbit carotid artery endarterectomy +anterior vein autogenous vein graft and postoperative phases of expression and the vein graft after exfoliative endometrial changes in segmental artery lumen cross-sectional area to study the HIF-1αand VEGF in a rabbit vein graft re-endothelialization role in the process.ObjectiveTo investigate the HIF-1αand VEGF expression in pairs of rabbits vein graft re-endothelialization of impact.1、HIF-la and VEGF in the rabbit carotid artery endarterectomy+anterior vein autogenous vein graft and postoperative phases of the expression of the relationship.2、after stripping of vein graft intimal changes in segmental artery lumen cross-sectional areaMethods1、group and the production of 6-month-old animal model 46 adult male New Zealand white rabbits were randomly divided into two groups, the first group of 10, the second group of 36 (including 30 for the trial to be animals, six of standby). Are based on ordinary food a week after feeding, collecting the first group of 10 rabbits were used in serum by enzyme-linked immunosorbent assay (ELISA) line of HIF-la and VEGF detection, and then take all 10 rabbits left cervical venous line HE staining do pathological examination to observe the arterial lumen area. The second group of 36 rabbits underwent left common carotid artery endarterectomy+ anterior vein autogenous vein graft, producing restenosis model,An ordinary diet after feeding.7 days after the operation,14 days,30 days followed by three time points were randomly selected from 10 rabbits collected serum by enzyme-linked immunosorbent assay (ELISA) line of HIF-la and VEGF testing, blood tests after taking the 10 rabbits in paragraph artery intima stripping HE staining did pathology laboratory, and to observe the arterial lumen area, the number of intimal hyperplasia and smooth muscle dynamics.2、pathological image analysis image analysis system using pathological specimens:Rabbit vein graft HE staining, Mason morphological analysis of stained films.3、chemical analysis of vein graft, respectively, prior to surgery and 7 days after 14 days,30 days, blood serum by ELISA measurement of HIF-1αand VEGF levels.Results1、HIF-la and VEGF expression in each period with a significant correlation (P <0.01); HIF-1αand VEGF in the rabbit autologous vein graft expression was significantly increased (P <0.01), in which the highest in 7 days, followed by a gradual decreased 30 days after the operation is only slightly higher than normal.2、HE staining showed intimal denudation arterial segment lumen area (LA) was the first overall diminishing, at the lowest value reached after 14 days and then gradually increasing trend.Conclusion1、this experiment has successfully produced a rabbit carotid artery endarterectomy and autogenous vein graft restenosis model.2、tissues and organs in acute hypoxia-ischemia period of HIF-la and VEGF expression increased rapidly; with prolonged ischemia and hypoxia of tissues and organs adapt gradually reduced two-factor concentrations, this experiment the observed HIF-la and VEGF expression peaked at 7 days after operation.3、after vein graft vascular remodeling in the early HIF-1αand VEGF can promote damage to the high expression of vascular endothelial cells (VEC) reconstruction of the structure and function to maintain the arterial lumen area; as time goes by, HIF-la and the the expression of VEGF gradually decreased after stripping of vein graft intimal segment arteries gradually narrowed to a minimum of 14 days after the operation, and then gradually increased.4、HIF-la and VEGF expression after stripping peaks and vein graft intimal minimum segment arteries, respectively after 7 days in 14 days, so after 7-14 days of HIF-la and VEGF expression in early reduction may be related to the vein graft restenosis.更多还原