College of Chemistry, Jilin Province Research Center for Engineering and Technology of Spectral Analytical Instruments, Jilin University, Qianjin Street 2699, Changchun, 130012, China.
College of Chemistry, Jilin Province Research Center for Engineering and Technology of Spectral Analytical Instruments, Jilin University, Qianjin Street 2699, Changchun, 130012, China; School of Pharmacy, Jilin University, Qianjin Street 2699, Changchun, 130012, China.
Talanta. 2024 Nov 1;279:126587. doi: 10.1016/j.talanta.2024.126587. Epub 2024 Jul 18.
The toxicity of organophosphorus pesticides (OPs) can catastrophically cause liver cell damage and inhibit the catalytic activity of cholinesterase. We designed and synthesized a near-infrared fluorescent probe HP-LZB with large Stokes shift which can specifically identify and detect butyrylcholinesterase (BChE) and visually explore the interaction between OPs and endogenous BChE in living cells. Fluorescence was turned on when HP-LZB was hydrolyzed into HP-LZ in the presence of BChE, and OPs could inhibit BChE's activity resulting in a decrease of fluorescence. Six OPs including three oxon pesticides (paraoxon, chlorpyrifos oxon and diazoxon) and their corresponding thion pesticides (parathion, chlorpyrifos and diazinon) were investigated. Both in vitro and cell experiments indicated that only oxon pesticides could inhibit BChE's activity. The limits of detection (LODs) of paraoxon, chlorpyrifos oxon and diazoxon were as low as 0.295, 0.007 and 0.011 ng mL respectively and the recovery of OPs residue in vegetable samples was satisfactory. Thion pesticides themselves could hardly inhibit the activity of BChE and are only toxic when they are converted to their corresponding oxon form in the metabolic process. However, in this work, thion pesticides were found not be oxidized into their oxon forms in living HepG2 cells due to the lack of cytochrome P450 in hepatoma HepG2 cell lines. Therefore, this probe has great application potential in effectively monitoring OPs in real plant samples and visually exploring the interaction between OPs and BChE in living cells.
有机磷农药（OPs）的毒性可能会灾难性地导致肝细胞损伤，并抑制胆碱酯酶的催化活性。我们设计并合成了一种具有大斯托克斯位移的近红外荧光探针 HP-LZB，它可以特异性识别和检测丁酰胆碱酯酶（BChE），并在活细胞中直观地探索 OPs 与内源性 BChE 之间的相互作用。当 HP-LZB 在存在 BChE 的情况下被水解成 HP-LZ 时，荧光被打开，而 OPs 可以抑制 BChE 的活性，导致荧光减弱。我们研究了六种 OPs，包括三种氧代农药（对氧磷、毒死蜱氧代和二嗪农氧代）及其相应的硫代农药（对硫磷、毒死蜱和地亚农）。体外和细胞实验均表明，只有氧代农药能够抑制 BChE 的活性。对氧磷、毒死蜱氧代和二嗪农氧代的检测限（LOD）分别低至 0.295、0.007 和 0.011 ng mL，且蔬菜样品中 OPs 残留的回收率令人满意。硫代农药本身很难抑制 BChE 的活性，只有在代谢过程中转化为相应的氧代形式时才具有毒性。然而，在这项工作中，由于肝癌 HepG2 细胞系中缺乏细胞色素 P450，我们发现硫代农药在活 HepG2 细胞中不会被氧化为其氧代形式。因此，该探针在有效监测实际植物样品中的 OPs 并直观地探索 OPs 与活细胞中 BChE 之间的相互作用方面具有巨大的应用潜力。
