静脉分子指纹Eph-B4在移植静脉适应动脉环境过程中的调控机制的动物模型研究

发布时间：2017-12-27 14:28

本文关键词：静脉分子指纹Eph-B4在移植静脉适应动脉环境过程中的调控机制的动物模型研究　出处：《昆明医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：[目的]将静脉移植到动脉环境中,静脉会在细胞或／和分子水平进行适应和调整,包括细胞增殖和细胞外基质(ECM)的沉淀或重塑,以使静脉壁增厚,在静脉移植过渡阶段产生病理性的血管内膜增生。Eph-B4在哺乳动物胚胎时期决定静脉系统的分化,并且,它被认为是成熟静脉系统的静态标记。此前进行的研究显示外科手术将静脉移植入动脉环境中是将静脉去特征化,其表现在Eph-B4的消失和管壁内膜的增厚。我们运用进行了静脉移植的大鼠动物模型来验证在静脉移植术后的静脉适应期间, Eph-B4对静脉管壁厚度起决定性作用的假设。[方法]我们将静脉移植至同种异体大鼠体内来建立静脉移植动物模型,以确定静脉在形态和生理上的变化过程。试验将供体鼠的肝上下腔静脉移植至受体鼠的肾下腹主动脉。将24只大鼠分为两组,其中12只入组对照组,12只入组试验组。两组都于移植后1周取出一半的大鼠体内的移植静脉,另一半的大鼠在移植后4周取出移植静脉。试验组需对移植静脉的Eph-B4进行刺激,在进行静脉移植术前将需移植的静脉在Ephrin-B2/Fc液中浸泡20分钟,并在静脉移植术后每天用Ephrin-B2/Fc液对大鼠进行腹腔注射直至试验终止取出静脉。所有的移植静脉均保存于液氮中,并对移植物标本进行HE染色、α-肌动蛋白免疫组化染色以及Eph-B4免疫荧光染色以研究标本在细胞及分子水平的变化。所有数据采用SPSS12.0软件分析和统计。[结果]随着静脉移植后4周静脉管壁的不断增厚,可见移植静脉管壁厚度的增加具有时间性。值得注意的是,相较于对照组4周时的移植静脉,经过Ephrin-B2/Fc液处理的试验组移植静脉管壁厚度减小并且α-肌动蛋白的表达减少。而试验1周所采集的样本两组间在移植静脉管壁厚度及α-肌动蛋白的表达上并无明显差别。还需注意的是,对照组移植静脉Eph-B4的表达减少并且在动脉环境中其静脉特征消失,而试验组经Ephrin-B2/Fc液处理的移植静脉其Eph-B4的表达得以保留。[结论]Eph-B4不只是成熟静脉系统的静态标记物,在静脉移植的过程中刺激Eph-B4可以保留静脉的特性并且防止静脉壁增厚。可见,持续对静脉标记物的刺激可以减少静脉移植的失败。刺激成熟细胞的Eph-B4为疾病的治疗提供了一条新的路径。
[Abstract]:[Objective] to the vein graft artery vein environment, will adapt and adjust the cell or / and molecular level, including cell proliferation and extracellular matrix (ECM) deposition or remodeling, in order to make the vein wall thickening in the vein graft intimal hyperplasia in transition stage of pathology. Eph-B4 determines the differentiation of the venous system during the mammalian embryo, and it is considered a static marker of the mature venous system. Previous studies have shown that surgical operation of venous transplantation into the arterial environment is characterized by venous disappearance, which is manifested by the disappearance of Eph-B4 and thickening of the inner wall of the tube. We used a rat model of venous transplantation to verify the hypothesis that Eph-B4 plays a decisive role in the thickness of venous wall during vein adaptation after vein transplantation. [Methods] we transplanted the vein to the allograft rats to establish the animal model of the vein graft to determine the morphological and physiological changes in the vein. The donor rat's superior and inferior vena cava was transplanted into the lower abdominal aorta of the recipient rat. 24 rats were divided into two groups, of which 12 were in the control group and 12 were in the test group. In the two groups, half of the rats were removed 1 weeks after transplantation, and the other half of the rats were taken out of the vein for 4 weeks after the transplantation. The experimental group needed to stimulate the Eph-B4 of the vein graft. Before vein transplantation, the vein needed to be transplanted was soaked for 20 minutes in the Ephrin-B2/Fc solution. After the intravenous transplantation, the rats were injected with Ephrin-B2/Fc solution every day until the end of the test. All the transplanted veins were stored in liquid nitrogen. HE staining, alpha actin immunohistochemical staining and Eph-B4 immunofluorescence staining were applied to study the changes of specimens at cellular and molecular level. All data are analyzed and counted by SPSS12.0 software. [results] with the continuous thickening of the venous wall 4 weeks after the vein transplantation, the thickness of the tube wall of the vein graft was more timeliness. It is worth noting that compared with the control group at 4 weeks, the thickness of transplanted venous wall decreased and the expression of alpha actin decreased in Ephrin-B2/Fc treated group. There was no significant difference in the thickness of the wall of the vein graft and the expression of alpha actin in the two groups of samples collected for 1 weeks. It is also necessary to note that the expression of Eph-B4 in the vein graft of the control group is reduced, and the vein characteristics disappeared in the arterial environment, while the expression of Eph-B4 in the experimental group treated with Ephrin-B2/Fc solution is preserved. [conclusion]Eph-B4 is not only a static marker of mature venous system, but also stimulates Eph-B4 during vein graft. It can retain the characteristics of vein and prevent thickening of vein wall. It can be seen that the continuous stimulation of the vein markers can reduce the failure of the vein graft. The Eph-B4 that stimulates the mature cells provides a new path for the treatment of disease.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R-332;R622.4

【相似文献】