CAS Key Laboratory of Coastal Environmental Processes and Ecological Remediation, Research Centre for Coastal Environmental Engineering and Technology, Yantai Institute of Coastal Zone Research , Chinese Academy of Sciences , Yantai 264003 , China.
University of Chinese Academy of Sciences , Beijing 100049 , China.
Anal Chem. 2018 Jul 3;90(13):8108-8115. doi: 10.1021/acs.analchem.8b01306. Epub 2018 Jun 12.
As important active sites of oxidoreductase in mitochondria, selenocysteine (Sec) takes the responsibility for cytoprotective effect and intracellular redox homeostasis. Carbon disulfide (CS) is a common solvent in industry, which can inhibit the activities of oxidoreductase and induce oxidative stress. It is necessary to investigate the cytoprotective effect of Sec against CS exposure. After integrated, the response moiety 2,4-dinitrobenzenesulfonamide and mitochondrial targeting moiety into the near-infrared heptamethine cyanine fluorophore, we develop a mitochondrial targeting near-infrared ratiometric fluorescent probe Mito- diNO for the selective and sensitive analysis of Sec concentration fluctuations in living cells and in mice models under the stimulation of CS. The probe can effectively accumulate in mitochondria and selectively detect the endogenous Sec concentrations in BRL 3A, RH-35, HL-7702, HepG2, and SMMC-7721 cell lines. The results indicate that CS exposure can lead to a decrease of Sec level and result in mitochondrial related acute inflammation. The exogenous supplement of Sec can protect cells from oxidative damage and reduce the symptoms of inflammation. We also establish CS induced acute and chronic hepatitis mice models to examine the tissue toxicity of CS and cytoprotection of Sec in liver. The organism can increase the concentration of Sec to deal with the damage caused by CS in acute hepatitis mice model. Also the exogenous supplement of Sec for the two mice models can effectively defend the CS induced liver damage. The real-time imaging of Sec concentrations in liver can be used to assess the degrees of liver injury during CS poisoning. The above applications make our probe a potential candidate for the clinical accurate diagnosis and treatment of CS poisoning.
作为线粒体中氧化还原酶的重要活性位点，硒代半胱氨酸 (Sec) 负责细胞保护作用和细胞内氧化还原稳态。二硫化碳 (CS) 是工业中常用的溶剂，它可以抑制氧化还原酶的活性并诱导氧化应激。因此，有必要研究 Sec 对 CS 暴露的细胞保护作用。我们将反应部分 2,4-二硝基苯磺酰胺和线粒体靶向部分整合到近红外庚烷氰染料中，开发了一种线粒体靶向近红外比率荧光探针 Mito- diNO，用于选择性和灵敏地分析活细胞和 CS 刺激下的小鼠模型中 Sec 浓度的波动。该探针可以有效地积聚在线粒体中，并选择性地检测 BRL 3A、RH-35、HL-7702、HepG2 和 SMMC-7721 细胞系中的内源性 Sec 浓度。结果表明，CS 暴露会导致 Sec 水平降低，并导致与线粒体相关的急性炎症。外源性补充 Sec 可以保护细胞免受氧化损伤，减轻炎症症状。我们还建立了 CS 诱导的急性和慢性肝炎小鼠模型，以研究 CS 在肝脏中的组织毒性和 Sec 的细胞保护作用。在急性肝炎小鼠模型中，机体可以增加 Sec 的浓度来应对 CS 造成的损伤。对于两种小鼠模型，外源性补充 Sec 可以有效地防御 CS 引起的肝损伤。肝脏中 Sec 浓度的实时成像可用于评估 CS 中毒期间肝脏损伤的程度。上述应用使我们的探针成为 CS 中毒临床准确诊断和治疗的潜在候选者。
