化学小分子和纳米金对自吞噬影响的研究

发布时间：2018-02-25 08:56

本文关键词： 金颗粒 3-MA Vps34　出处：《清华大学》2012年博士论文　论文类型：学位论文

【摘要】：自噬是相对保守机制，它通过包裹细胞内受损的细胞器以及错误折叠的蛋白质将其运输到溶酶体中降解以维持细胞的动态平衡。自噬在癌症和神经退行性疾病中有着非常重要的作用。3-MA是应用最为广泛的自吞噬体的抑制剂。但其溶解度非常差，100%抑制自吞噬体的浓度为10mM。3-MA（3-methyladenine）为6-氨基-3-甲基嘌呤，它是由嘧啶和咪唑环并合而成的并环体系。而3-MA N3和C6位是活性官能团，从氨基出发改造最为容易，，可以形成多种极性的基团。我们通过在3-MA的N3和C6加入不同的官能团合成出36种3-MA的衍生物。经过高通量筛选发现有3种化学小分子能够抑制自噬。应用免疫荧光进一步验证了20,24和34在浓度为1mM对自吞噬体有非常好的抑制效果，特别是34100%抑制自噬的最低浓度为50μM。并确定这三种化学小分子抑制自吞噬是通过抑制Vps34酶的活性实现的。我们研究表明对3-MA改造是开发自吞噬体抑制剂的最有效的方式，并为临床研究奠定基础。最近研究表明纳米颗粒诱导自吞噬。金颗粒是一种纳米材料，在生物学和医学方面应用的非常广泛。我们通过实验发现，金颗粒通过内吞途径进入细胞，且直径越大进入细胞越多，最终滞留在溶酶体中，并通过改变溶酶体pH值破坏了溶酶体使自吞噬的不能被溶酶体降解，导致细胞中自吞噬体增多。我们的研究为纳米材料在生物学及医学中的应用有着非常重要的指导作用。
[Abstract]:Autophagy is a relatively conservative mechanism, It is transported into lysosomes by encapsulating damaged organelles and misfolded proteins in order to maintain the dynamic balance of cells. Autophagy plays a very important role in cancer and neurodegenerative diseases .3-MA. Is the most widely used inhibitor of autophagocytosis, but its solubility is very poor and its concentration of 100% inhibition of autophagocytosis is 10mM.3-MA-3-methyladenine (6-amino-3-methyladenine). It is a concyclic system composed of pyrimidine and imidazole rings, while 3-MA N 3 and C 6 are active functional groups, which are most easily modified from amino groups. 36 derivatives of 3-MA were synthesized by adding different functional groups into 3-MA N3 and C6. Three kinds of chemical small molecules were found to inhibit autophagy by high-throughput screening. The light further demonstrated that 20 and 34 had a very good inhibitory effect on the phagocytosis at a concentration of 1 mm. In particular, the minimum concentration of 34100% in inhibiting autophagy was 50 渭 M. and it was determined that the inhibition of autophagy by these three chemical small molecules was achieved by inhibiting the activity of Vps34 enzyme. Our study shows that 3-MA modification is the most effective way to develop autophagocytic inhibitors. And lay the foundation for clinical research. Recent studies have shown that nanoparticles are induced autophagocytosis. Gold particles are a kind of nanomaterials that are widely used in biology and medicine. The gold particles enter the cells through endocytosis pathway, and the larger the diameter is, the more the gold particles enter into the lysosome, and finally they stay in the lysosome, and destroy the lysosome by changing the pH value of the lysosome so that the self-phagocytosis can not be degraded by the lysosome. Our research is very important for the application of nanomaterials in biology and medicine.
【学位授予单位】：清华大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R363

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