抵抗素调控血管内皮细胞对葡萄糖和胰岛素转运的研究

发布时间：2018-05-02 07:18

本文选题：抵抗素 + 血管内皮细胞　；参考：《华中农业大学》2015年硕士论文

【摘要】：血液中的葡萄糖和胰岛素透过血管壁进入组织器官是维持人体葡萄糖稳态的重要步骤,若葡萄糖和胰岛素滞留在血管内无法到达组织发挥功能,则将导致机体的高血糖、胰岛素抵抗以及二型糖尿病。血管壁由单层或多层血管内皮细胞构成,血管内皮细胞对葡萄糖和胰岛素的转运能力是葡萄糖和胰岛素能否到达组织器官的关键。抵抗素自发现以来一直被认为与胰岛素抵抗、二型糖尿病相关。前期研究发现,抵抗素处理后的血管内皮细胞中葡萄糖和胰岛素的通透性下降,预示着抵抗素通过一条新的通路导致机体胰岛素抵抗,而这一作用的分子机制还未知。为了探究抵抗素降低血管内皮细胞对葡萄糖和胰岛素的通透性的作用机制,本研究以人脐静脉瘤融合细胞(EA.hy 926)为体外血管壁模型,检测了血管内皮细胞转运葡萄糖和胰岛素必须的两种因子GLUT1和INSR的表达水平,并构建了GLUT1启动子载体,利用海肾双荧光报告系统检测了GLUT1启动子的转录活性,探讨了抵抗素调控GLUT1表达的分子机制。获得结果如下:1.用抵抗素刺激EA.hy 926细胞,RT-PCR和western blot的检测发现,抵抗素可下调GLUT1和INSR的m RNA水平以及蛋白水平。2.免疫荧光试验显示,抵抗素刺激使EA.hy 926细胞GLUT1蛋白的荧光强度减少;而超表达GLUT1可回复抵抗素引起的葡萄糖通透性下调。3.为了探索抵抗素下调GLUT1的机制,构建了GLUT1启动子载体,双荧光报告系统检测发现,PPARγ可特异的与GLUT1启动子结合,上调启动子转录活性,表明抵抗素是通过PPARγ调控GLUT1的表达。研究证明,抵抗素通过PPARγ—GLUT1—葡萄糖转运信号通路调控葡萄糖在血管内皮细胞中的转运,从而影响葡萄糖进入血管内皮细胞,减少组织对葡萄糖的氧化利用。
[Abstract]:Glucose and insulin in the blood flow through the wall of the blood vessels are important steps to maintain the body's glucose homeostasis. If glucose and insulin are retained in the blood vessels that can not reach the tissue and function, it will lead to hyperglycemia, insulin resistance, and type two diabetes. The vascular walls are composed of single or multilayer vascular endothelial cells. The transport capacity of vascular endothelial cells to glucose and insulin is the key to the ability of glucose and insulin to reach the organs of the tissue. Resistin has been found to be associated with insulin resistance, type two diabetes since its discovery. Previous studies have found that the permeability of glucose and insulin in vascular endothelial cells after resistin treatment has been found. In order to explore the mechanism of the effect of resistin on the permeability of glucose and insulin in vascular endothelial cells, this study uses the fusion cell of human umbilical vein aneurysm (EA.hy 926) as an in vitro vascular wall model to detect blood. The expression level of two factors GLUT1 and INSR must be transported by endothelial cells, and the GLUT1 promoter was constructed. The transcriptional activity of GLUT1 promoter was detected by the double fluorescence report system of the sea kidney, and the molecular mechanism of resistin to regulate the expression of GLUT1 was discussed. The results were as follows: 1. the EA.hy 926 finer was stimulated with resistin. The test of cell, RT-PCR and Western blot found that resistin could downregulate the m RNA level of GLUT1 and INSR and protein level.2. immunofluorescence test that resistin stimulation reduced the fluorescence intensity of GLUT1 protein in EA.hy 926 cells, while the overexpression of the GLUT1 could restore the downregulation of glucose induced by resistin in order to explore the downregulation of resistin. 1 mechanism, the GLUT1 promoter was constructed, and the double fluorescence report system detected that PPAR gamma could be specifically associated with GLUT1 promoter, up regulation of promoter transcriptional activity, indicating that resistin was regulated by PPAR gamma regulation of GLUT1 expression. The study showed that resistin was regulated by PPAR gamma GLUT1 glucose transport signaling pathway to regulate glucose in vascular endothelium Transport in cells affects glucose entry into vascular endothelial cells and reduces the use of glucose in tissues.

【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

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