基于四苯基乙烯的聚集诱导发光高分子的合成及其在基因递送体系中的应用

发布时间：2018-06-14 14:15

本文选题：基因转染 + 聚甲基丙烯酸缩水甘油酯　；参考：《天津理工大学》2016年硕士论文

【摘要】：近年来,阳离子高分子由于具有合成简单、价格便宜、生物相容性好、生物可降解、无毒无免疫原性、水溶性好等性质,在基因递送体系中得到了广泛的应用。本课题中,我们首先合成了两种不同种类的高分子:一种是通过原子转移自由基聚合(ATRP)方法得到的线性聚甲基丙烯酸缩水甘油酯,并对其进行了连续的环糊精、乙二胺和胍基化修饰;另一种是对枝状的聚乙烯亚胺进行环糊精修饰,并通过核磁共振氢谱和傅里叶红外光谱验证了两种高分子的成功合成。在电荷作用下,分别与pDNA形成了复合体,琼脂糖凝胶电泳实验表明两种高分子都能够充分的保护pDNA,扫描显微镜结果表明复合体都是均一的球形纳米粒子,并且都表现出了较低的细胞毒性和较高的转染效率。此外,胍基化的修饰能够有效提高复合体的溶酶体逃逸率和基因转染效率。为达到追踪基因释放过程的目的,我们将具有聚集诱导发光(AIE)性质的分子引入到了基因递送体系中。我们首先合成了具有顺式结构的双硼酸基团修饰的四苯基乙烯(TPEDB),然后将其分别与两种高分子作用,通过硼酸酯键的形成得到了两种聚集诱导发光高分子。其次,我们分别对两种AIE高分子与pDNA作用形成的复合体进行了表征,它们都能够很好地保护pDNA,并形成了更小的纳米粒子,并且能够提高纳米粒子的稳定性。激光扫描共聚焦实验结果表明:AIE高分子具有更好的基因递送能力,甚至能够将pDNA运送至细胞核中。通过将荧光染料NBD标记的DNA和AIE高分子构建成荧光共振能量转移体系,在双光子激光扫描共聚焦下,我们能够更好地追踪pDNA的释放过程及载体、pDNA的细胞内分布,为多功能基因递送体系的设计提供了一种新的方法。
[Abstract]:In recent years, cationic polymers have been widely used in gene delivery systems due to their advantages of simple synthesis, low cost, good biocompatibility, biodegradability, non-toxic and non-immunogenicity, and good water solubility. In this paper, we first synthesized two kinds of polymers: one is linear poly (glycidyl methacrylate) obtained by atom transfer radical polymerization (ATRP), and the continuous cyclodextrin was obtained. Ethylenediamine and guanidine were modified with cyclodextrin, and the two polymers were successfully synthesized by NMR and Fourier transform infrared spectroscopy (FTIR). Under the action of electric charge, the complex was formed with pDNA respectively. Agarose gel electrophoresis showed that both polymers could protect pDNA. scanning microscope showed that the complexes were homogeneous spherical nanoparticles. All of them showed low cytotoxicity and high transfection efficiency. In addition, guanidine modification can effectively improve the lysosomal escape rate and gene transfection efficiency of the complex. In order to track the gene release process, we introduced the agglomeration induced luminescence (AIEs) molecules into the gene delivery system. We first synthesized tetraphenylethylene modified with cis-diborate group and then reacted with two kinds of polymers respectively. Two kinds of aggregation induced luminescent polymers were obtained by the formation of borate ester bonds. Secondly, we characterize the complexes formed by the interaction of two AIE polymers with pDNA. They can protect pDNAs, form smaller nanoparticles and improve the stability of nanoparticles. The results of laser scanning confocal experiments show that the molecular weight of 1: AIE has better gene delivery ability and can even transport pDNA into the nucleus. The fluorescence resonance energy transfer system was constructed by using fluorescent dye NBD labeled DNA and AIE polymer. Under the condition of two-photon laser scanning confocal, we can better track the release process of pDNA and the intracellular distribution of pDNA vector. It provides a new method for the design of multifunctional gene delivery system.
【学位授予单位】：天津理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O631.24

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