超声介导微泡造影剂对细胞和微血管以及基因在HUVEC内转染率的影响的研究

发布时间：2018-04-14 16:00

本文选题：微泡造影剂 + 人脐静脉内皮细胞　；参考：《中国医科大学》2006年博士论文

【摘要】：前言随着分子生物学的发展和基因工程技术的改进,基因治疗的研究不断取得进步。超声微泡造影剂(USMBCA)的应用使得超声医学不但在诊断方面有所创新,而且其在基因治疗方面潜在的应用价值倍受关注,具有广阔的应用前景。基因治疗时治疗基因必须到达靶细胞,进入细胞核,然后再经过转录、翻译为蛋白质,才能起到治疗作用。基因导入体内的方法有直接注射和静脉注射法。如果不用某些方式使这些治疗性基因保持稳定,经静脉注射后通常会被血清酶快速清除,或很难通过内皮细胞屏障。在静脉给药途径中,要防止基因物质在血液中被降解。人们已经尝试用各种不同的被覆物质作为基因的载体来改善基因物质的转移,新近研究发现,USMBCA可成为基因治疗的一种安全、有效的载体。但是,如果使用超声剂量过大,或微泡造影剂(MBCA)浓度过高时,利用超声介导MBCA实现基因的定向转移这一技术同样也会对机体产生一定的有害的生物学效应。影响USMBCA介导基因治疗效果的因素很多,除不同组织和基因种类的影响外,超声能量、辐照时间、基因与微泡比例等诸多因素均可对基因治疗产生影响。基础研究的意义是为了临床应用,虽然在基因治疗中超声介导微泡可通过空化效应增加内皮细胞间隙和细胞膜的通透性,从而使治疗基因进入靶细胞;但同时也可能引起一定的组织损伤。因此,为了该项技术在临床中安全有效地应用,对超声介导MBCA各种相关的参数进行深入的研究是十分必要的。本研究旨在:通过观察超声介导MBCA对体外培养的人脐静脉内皮细胞(HUVEC)的作用,静脉注射USMBCA对大鼠心肌组织毛细血管及肌纤维的作用,以及使用超声介导MBCA对绿色荧光蛋白报告基因(EPGFP)对HUVEC的转染效率的影响,探讨USMBCA是否可以作为基因治疗的一种
[Abstract]:PrefaceWith the development of molecular biology and the improvement of genetic engineering technology, the research of gene therapy has made progress.The application of ultrasound microbubble contrast agent USMBCAmakes ultrasound medicine not only be innovated in diagnosis, but also its potential application value in gene therapy has attracted much attention and has a broad application prospect.In gene therapy, the therapeutic gene must reach the target cell, enter the nucleus, and then be transcribed and translated into protein to play a therapeutic role.Direct injection and intravenous injection are the methods of gene transfer in vivo.If these therapeutic genes are not stabilized in some way, they are usually quickly cleared by serum enzymes after intravenous injection, or are difficult to pass through the endothelial cell barrier.During intravenous administration, genetic substances are prevented from degrading in the blood.People have tried to use a variety of coated materials as gene carriers to improve gene transfer. Recently, USMBCA has been found to be a safe and effective vector for gene therapy.However, if the dose of ultrasound is too high or the concentration of microbubble contrast agent is too high, the technology of directed gene transfer by ultrasound mediated MBCA will also produce some harmful biological effects on the body.There are many factors that affect the effect of gene therapy mediated by USMBCA. In addition to different tissues and gene types, ultrasound energy, irradiation time, gene / microbubble ratio and many other factors can affect gene therapy.The significance of basic research is for clinical application, although ultrasound mediated microbubbles can increase the permeability of endothelial cell space and cell membrane through cavitation effect in gene therapy, so that therapeutic gene can enter target cells.But at the same time, it may cause some tissue damage.Therefore, in order to safely and effectively apply this technique in clinic, it is necessary to study various parameters related to ultrasound mediated MBCA.The purpose of this study was to observe the effects of ultrasound mediated MBCA on cultured human umbilical vein endothelial cells (HUVECs) and the effects of intravenous USMBCA on the capillaries and muscle fibers in rat myocardium.The effect of ultrasound mediated MBCA on the transfection efficiency of green fluorescent protein reporter gene (EGFP) on HUVEC was also discussed. Whether USMBCA could be used as a gene therapy was discussed.
【学位授予单位】：中国医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R346

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