硫醇螢光探針之設計與合成及活體細胞的應用

Abstract

此篇論文中設計與合成出螢光感測器NBDMP可以偵測小分子硫醇Cys、Hcy和GSH。 NBDMP是由硝基苯噁二唑與吡啶陽離子兩個螢光團組成的螢光感測器，用來偵測生物分子Cys、Hcy及GSH。硝基苯噁二唑作為小分子硫醇的感測端，當與(E)–4-(4–hydroxystyryl)-1-methylpyridinium iodide結合時，會使螢光團的螢光淬滅。NBDMP與小分子硫醇反應後，螢光淬滅效應消失，產生10倍的綠色螢光增強且Cys、Hcy與硝基苯噁二唑反應時，會發生分子內重排，分別產生的Cys-NBD及Hcy-NBD的硝基苯噁二唑黃色螢光。NBDMP具有良好的選擇性與靈敏性，並可以以不同螢光反應區分GSH與Cys／Hcy，它對Cys、Hcy及GSH的偵測極限分別為73 nM、51nM及128 nM，並可應用於細胞中生物硫醇顯影當中。

In this thesis, fluorescent probe NBDMP is designed and synthesized to detect biothiols Cys, Hcy, GSH. The fluorescent probe NBDMP consisted of two fluorophores NBD and (E)–4-(4–hydroxystyryl)-1-methyl pyridinium iodide. NBD is a fluorophore and also a reaction part for thiol. NBDMP reacts with Cys, Hcy, GSH to obtain 10-fold green fluorescent enhancement from pyridinium fluorophore and 65-fold and 110-fold yellow fluorescent enhancement from NBD fluorophore by Cys and Hcy due to intramolecular sulfur-amine rearrangement in NBD fluorophore. NMDMP also shows good selectivity to distinguish Cys/Hcy from GSH. The detection limit of Cys, Hcy and GSH are 73 nM, 51nM and 128 nM, respectively and can be used in biothiol imaging.