超声联合超声造影剂促进VEGF基因转染的体内、外研究

发布时间：2018-06-05 03:01

本文选题：基因治疗 + 成纤维细胞　；参考：《吉林大学》2006年博士论文

【摘要】： 基因治疗有可能成为人类最终征服许多难治性疾病的一种新的治疗手段。如何将目的基因安全、高效、靶向性地导入人体内特定器官、组织,并在靶细胞内表达是目前有待解决的难题之一。我们观察了成纤维细胞在诊断剂量的超声波及联合应用超声造影剂作用下的电镜改变,发现细胞膜表面形成一些可逆性的小孔。说明诊断剂量的超声波联合超声造影剂对细胞的损害作用是轻微的、可逆的。我们观察了诊断剂量范围不同超声强度对体内外基因转染的影响,得出如下结论:MI(mechanism index)为1.4的超声剂量与超声造影剂联合适合作用于体外培养细胞;而超声联合造影剂应用于体内细胞的基因转染时,MI为1.6的超声剂量比较合适。在转染后成纤维细胞血管内皮细胞生长因子(Vascular endothelial cell growth factor)VEGF165蛋白表达的定量研究中,发现超声联合造影剂不但可促进基因的转染,对基因的表达也有促进作用。体内部分实验,我们设计了大鼠背部随意皮瓣的动物模型,结果表明,超声联合造影剂能够促进VEGF基因的体内转染,使缺血皮瓣毛细血管生成增多,皮瓣成活面积增加。通过本实验说明诊断剂量超声联合超声造影剂是一种安全、有效、简便的促进基因转染的方法。把超声联合造影剂这种新的转基因方法扩展到组织移植方面的研究中来,为创伤修复提供了一种简捷、高效的途径。
[Abstract]:Gene therapy may become a new method to conquer many refractory diseases. How to transfer the target gene safely, efficiently and targeted into specific organs and tissues and express it in target cells is one of the difficult problems to be solved. We observed the electron microscopic changes of fibroblasts under the action of diagnostic dose of ultrasound and ultrasound contrast medium, and found that some reversible pores were formed on the surface of cell membrane. The results showed that the damage of diagnostic dose of ultrasound combined with ultrasound contrast agent on cells was mild and reversible. We observed the effects of different ultrasound intensities on gene transfection in vitro and in vivo. The conclusions are as follows: the ultrasound dose of 1. 4% MImechanism and ultrasound contrast agent are suitable for cell culture in vitro. However, ultrasound combined with contrast agent was more suitable for gene transfection with an ultrasound dose of 1.6. In the quantitative study on the expression of vascular endothelial cell growth factor)VEGF165 protein of vascular endothelial growth factor (VEGF) in fibroblasts after transfection, it was found that ultrasound combined with contrast agent could not only promote gene transfection, but also promote gene expression. In some experiments in vivo, we designed an animal model of random dorsal flap in rats. The results showed that ultrasound combined with contrast agent could promote the transfection of VEGF gene in vivo, increase capillary formation and increase the survival area of ischemic flap. The results show that the diagnostic dose ultrasound combined with ultrasound contrast agent is a safe, effective and simple method to promote gene transfection. The application of ultrasound combined with contrast agent to the study of tissue transplantation provides a simple and efficient way for wound repair.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R346

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