高選擇性超氧化物螢光化學感測分子之設計與合成及於活體細胞的應用

Abstract

本論文以BODIPY和coumaroin為螢光基團主體，修飾上雙苯基磷官能基作為辨識端，設計並合成出螢光化學感測分子TPB和DPC。此系列的螢光化學感測分子皆可以利用超氧化物的強氧化力來與辨識端反應，使原本沒有任何螢光放射的TPB和DPC，分別產生綠色及藍色的螢光。二者在多種活性氧化物及氮化物下，對於過氧化物具有高度的選擇性及靈敏性，只有在超氧化物存在下，才會有明顯的螢光強度變化，達到偵測的效果。 最後，將TPB和DPC應用於細胞顯影實驗中，在老鼠巨噬細胞(Raw 264.7)中，利用共軛焦顯微鏡，成功觀測到TPB和DPC對超氧化物的偵測能力，並可更進一步偵測由PMA刺激所產生的超氧化物。最後，由細胞毒性檢可知，當TPB和DPC之濃度達20 μM時，巨噬細胞的存活率仍超過80 %。

In this thesis, BODIPY-based and coumaroin-based fluorescent chemosensors TPB and DPC were designed and synthesized for detection of superoxide. The design of fluorescent probes is to join a superoxide-reactive moiety with an organic fluorophore. The reaction of probes with superoxide produces significant fluorescence enhancement. TPB and DPC displayed good selectivity and sensitivity for superoxide over other ROS (reactive oxygen species) and RNS (reactive nitrogen species). In addition, confocal fluorescence microscopy imaging using RAW264.7 macrophages demonstrates that the probe TPB and DPC could be an efficient fluorescent probe for superoxide in living cells. According to cytotoxicity tests, cell viabilities were above 80 %, when the concentrations of TPB and DPC were 20 μM.