新型心肌黄酶检测荧光探针的合成及生物成像  

Synthesis and bio-imaging application of novel fluorescent probes for the detection of diaphorase

作　　者：周莹 房街芹 贾明轩 朱立平 ZHOU Yin;FANG Jieqin;JIA Mingxuan;ZHU Liping(College of Chemical Science and Technology,Yunnan University,Kunming 650091,China)

机构地区：[1]云南大学化学科学与工程学院,云南昆明650091

出　　处：《南昌大学学报（理科版）》2022年第1期72-78,84,共8页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(22067019)。

摘　　要：由于癌细胞以极快的速度不停生长,其内部需氧量远远超出血液的供应量,导致组织内部出现低氧。在固体肿瘤内,低氧导致部分酶出现过表达现象,其中硝基还原酶、偶氮还原酶及心肌黄酶的浓度变化最为明显。本研究中,我们以心肌黄酶作为检测目标,合成了一种新型荧光探针Milla 1。该探针将苯醌作为反应基团,在还原型烟酰胺腺嘌呤二核苷酸的共同作用下,经过还原反应并发生质子化,生成对苯二酚(或半醌),利用断键反应释放出荧光团,实现对心肌黄酶的光学检测。其中,Milla 1可与心肌黄酶发生专一识别反应,通过635 nm处荧光值的变化直接测定心肌黄酶含量的波动,进而检测出相关低氧区域的低氧程度。由于探针Milla 1具有选择性高、光稳定性好、毒性低等特点,探针Milla 1被成功用于HeLa细胞和秀丽隐杆线虫内源性和外源性的心肌黄酶检测和光学成像实验中。Because cancer cells grow at an extremely fast rate,their internal oxygen demand far exceeds the blood supply,resulting in hypoxia in the tissues.In solid tumors,hypoxia leads to over-expression of some enzymes,among which the concentrations of nitroreductase,azo reductase and diaphorase change the most.In this study,we took diaphorase as the detection target and synthesized a novel fluorescent probe Milla 1,with the benzoquinone as a recognition group.With nicotinamide adenine dinuclectide,it underwent a reduction reaction to generate hydroquinone(or semiquinone)and released the fluorophore through a bond-breaking reaction,achieving the optical detection effects towards diaphorase.In the specific recognition reaction,the diaphorase content fluctuation was directly detected by the changes of fluorescence values at 635 nm,and then the hypoxia degree in the relevant hypoxic area was deduced.Because probe Milla 1 has the advantages of high selectivity,good photostability,and low toxicity,it was successfully used in detection and optical imaging experiments of endogenous and exogenous diaphorase in HeLa cells and Caenorhabditis elegans.

关 键 词：荧光探针 心肌黄酶 三甲基苯醌 细胞成像 

分 类 号：O652[理学—分析化学]