Synthesis of an ultrasensitive BODIPY-derived fluorescent probe for detecting HOCl in live cells
Synthesis of an ultrasensitive BODIPY-derived fluorescent probe for detecting HOCl in live cells
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DOI：
10.1038/s41596-018-0041-6
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发表时间：
2018-09
期刊：
影响因子：
14.8
通讯作者：
Hao Zhu;Zhen Zhang;Saran Long;Jianjun Du;Jiangli Fan;Xiaojun Peng
中科院分区：
文献类型：
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作者：
Hao Zhu;Zhen Zhang;Saran Long;Jianjun Du;Jiangli Fan;Xiaojun Peng
次氯酸（HOCl）是由免疫细胞产生以对抗感染的活性氧（ROS）的关键成员。另一方面，HOCl在体内平衡中引起生物分子的氧化损伤，并与许多疾病有关，包括炎症、神经退行性疾病和心血管疾病。在此，我们详细介绍了一种硼-二吡咯亚甲基（BODIPY）衍生的荧光探针HOCl（BClO）的制备过程及其作为活细胞成像试剂的应用。BClO是通过四步程序在一锅中合成的，除了起始原料的比例之外，该程序与常规BODIPY染料的制备几乎相同。BClO具有非常快速的反应（在几秒钟内饱和），对HOCl非常敏感。BClO的检测限达到了亚纳摩尔范围，这是迄今为止最高的HOCl灵敏度。利用BClO的超灵敏特性，我们先前已经证明了其检测由巨噬细胞产生的内源性HOCl的能力，并且表明其也可以用于区分癌细胞系（其显示高HOCl产生）与非癌细胞系（其显示低HOCl产生）。该方案需要约2天的探针合成和长达约18小时的荧光成像和流式细胞术测定。
Hypochlorous acid (HOCl) is a critical member of the reactive oxygen species (ROS) produced by immune cells to fight infections. On the other hand, HOCl in homeostasis causes oxidative damage to biomolecules and is linked to many diseases, including inflammatory, neurodegenerative, and cardiovascular diseases. Herein, we detail a procedure for the preparation of a boron-dipyrromethene (BODIPY)-derived fluorescent probe for HOCl (BClO) and its application as an imaging reagent in living cells. BClO is synthesized in one pot through a four-step procedure that is nearly the same as that for conventional BODIPY dye preparation, except for the ratio of starting materials. BClO has an extremely rapid response (saturated within seconds) and is ultrasensitive to HOCl. The detection limit of BClO reaches the subnanomolar range, which is the highest HOCl sensitivity to date. Taking advantage of the ultrasensitive character of BClO, we have previously demonstrated its ability to detect endogenous HOCl generated by macrophages and shown that it can also be used to discriminate cancer cell lines (which show high HOCl production) from non-cancer cell lines (which show low HOCl production). The protocol requires ~2 d for probe synthesis and up to ~18 h for fluorescence imaging and flow cytometry assays.