测定ClO~-的水溶性荧光探针的合成及应用
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摘要
生物体内存在的大量活性氧(ROS)等小分子物质在生命活动过程中起着抗肿瘤、抗炎、抗衰老等极为重要的作用。其中ClO~-是一类重要的活性氧,适量的ClO~-可以产生有效的抗菌作用;但生物体内ClO~-过度积累可导致组织损伤,引发多种疾病。因此,快速灵敏、原位检测生物体内ClO~-的浓度具有重要的研究意义。该论文设计并合成了以对甲氧基苯酚为识别基团的基于香豆素骨架的新型荧光探针HMAT。探针HMAT本身在500nm处有微弱的荧光信号,加入ClO~-后,探针在500 nm处的荧光信号显著增强,反应体系的荧光强度与ClO~-浓度在1. 0～9. 0μmol/L内呈现良好的线性关系(R2=0. 993 2),最低检出限为1. 3 nmol/L,在神经母瘤细胞SH-SY5Y中具有良好的荧光成像效果。
A large number of small molecules in living organisms play an important role in anti-tumor,anti-inflammatory and anti-aging functions in the process of life activities such as reactive oxygen species( ROS). Among them,ClO~- is a vital class of ROS,and an appropriate amount of ClO~- could produce an effective antibacterial effect; however,excessive accumulation of ClO~- in the body could cause tissue damage and various diseases. Therefore,rapid and sensitive detection for the concentration of ClO~- in vivo has important research significance. In this paper,a novel fluorescent probe HMAT based on coumarin skeleton was designed and synthesized using p-methoxyphenol as a recognition group. Probe HMAT has a weak fluorescence signal at 500 nm,the fluorescence intensity of the probe at 500 nm is significantly enhanced after adding ClO~- ,and the fluorescence intensity of the reaction system and the ClO~- concentration are an excellent linear relationship in the range of 1. 0 ～ 9. 0 μmol/L( R~2= 0. 993 2) and a low detection limit of 1. 3 nmol/L. Moreover,the probe has a good fluorescence imaging effect in neuroblastoma SH-SY5 Y with negligilble cytotoxicity.
A large number of small molecules in living organisms play an important role in anti-tumor,anti-inflammatory and anti-aging functions in the process of life activities such as reactive oxygen species( ROS). Among them,ClO~- is a vital class of ROS,and an appropriate amount of ClO~- could produce an effective antibacterial effect; however,excessive accumulation of ClO~- in the body could cause tissue damage and various diseases. Therefore,rapid and sensitive detection for the concentration of ClO~- in vivo has important research significance. In this paper,a novel fluorescent probe HMAT based on coumarin skeleton was designed and synthesized using p-methoxyphenol as a recognition group. Probe HMAT has a weak fluorescence signal at 500 nm,the fluorescence intensity of the probe at 500 nm is significantly enhanced after adding ClO~- ,and the fluorescence intensity of the reaction system and the ClO~- concentration are an excellent linear relationship in the range of 1. 0 ～ 9. 0 μmol/L( R~2= 0. 993 2) and a low detection limit of 1. 3 nmol/L. Moreover,the probe has a good fluorescence imaging effect in neuroblastoma SH-SY5 Y with negligilble cytotoxicity.
引文
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[2] SHEN S L,ZHANG X F,GE Y Q,et al. A novel ratiometric fluorescent probe for the detection of HOCl based on FRET strategy[J]. Sensors and Actuators B:Chemical,2018,254:736-741.
[3] LIN Q S,HUANG Y L,FAN X X,et al. A ratiometric fluorescent probe for hypochlorous acid determination:Excitation and the dual-emission wavelengths at NIR region[J]. Talanta,2017,170:496-501.
[4] ZHANG Y R,CHEN X P,SHAO J,et al. A ratiometric fluorescent probe for sensing HOCl based on a coumarin-rhodamine dyad[J]. Chemical Communications,2014,50(91):14241-14244.
[5] ZHANG Y R,ZHAO Z M,MIAO J Y,et al. A ratiometric fluorescence probe based on a novel FRET platform for imaging endogenous HOCl in the living cells[J]. Sensors and Actuators B:Chemical,2016,229(28):408-413.
[6] ALBERS A E,OKREGLAK V S,CHANG C J,et al.A FRET-based approach to ratiometric fluorescence detection of hydrogen peroxide[J]. Journal of the American Chemical Society,2006,128(30):9640-9641.
[7] MICHAEL C V. Efficient synthesis of the gossypol binaph backbone[J]. Journal of Organic Chemistry,1981,46(15):3124-3127.
[8] CABRERA E V,GIL A C,ORTEGA J G. A one-pot synthesis of 6-iso-propyl-7-methoxy-1-etralone and 6-isopropyl-7-me-thoxy-2-tetralone[J]. Organic Preparations and Procedures International,2014,46(3):272-276.
[9] CHRISTOS I S,EFSTRATIOS N,THEOCHARIS V K. A scalable synthesis of the antidepressant agomelatine by a tandem allylic chlorination-isomerization process[J]. European Journal of Organic Chemistry,2014,46(29):6376-6379.
[10] BABIOR B M. Nadph oxidase[J]. Current Opinion in Immunology,2004,16(1):42-47.
[11] ZGLICZYNSKI J M, STELMASZYNSKA T, OSTROWSKI W,et al. Myeloperoxidase of human leukaemic leucocytes[J]. Febs Journal,1968,4(4):540-547.
[12] HAMMERSCHMIDT S,BUCHLER N,WAHN H.Tissue lipid peroxidation and reduced glutathione depletion in hypochlorite-induced lung injury[J].Chest,2002,121(2):573-581.
[13] WEITZMAN S A,GORDON L I. Inflammation and cancer:Role of phagocyte-generated oxidants in carcinogenesis[J]. Blood,1990,76(4):655-663.
[14] MALLE E,BUCH T,GRONE H J. Myeloperoxidase in kidney disease[J]. Kidney International,2003,64(6):1956-1967.
[15] HASEQAWA T,MALLE E,FARHOOD A,et al.Generation of hypochlorite-modified proteins by neutrophils during ischemia-reperfusion injury in rat liver:Attenuation by ischemic preconditioning[J]. American Journal of Physiology Gastrointestinal and Liver Physiology,2005,289(4):760-767.
[16] DAUGHERTY A,DUNN J L,RATERI D L,et al.Myeloperoxidase,a catalyst for lipoprotein oxidation,is expressed in human atherosclerotic lesions[J].Journal of Clinical Investigation,1994,94(1):437-444.
[17] HAZELL L J,ARNOLD L,FLOWERS D,et al.Presence of hypochlorite-modified proteins in human atherosclerotic lesions[J]. Journal of Clinical Investigation,1996,97(6):1535-1544.
[18] PODREZE E A,ABU-SOUD H M,HAZEN S L. Myeloperoxidase-generated oxidants and atherosclerosis[J]. Free Radical Biology and Medicine,2000,28(12):1717-1725.
[19] HAZEN S L,HEINECKE J W. 3-Chlorotyrosine,a specific marker of myeloperoxidase-catalyzed oxidation,is markedly elevated in low density lipoprotein isolated from human atherosclerotic intima[J]. Journal of Clinical Investigation,1997,99(9):2075-2081.
[20] NING Y Y,WANG X Q,SHENG K J,et al. A novel colorimetric and fluorescence turn-on p H sensor with a notably large stokes shift for its application[J]. New Journal of Chemistry,2018,42:14510-14516.
[21] CHEN H,TANG Y H,REN M G,et al. Single nearinfrared fluorescent probe with high and low-sensitivity sites for sensing different concentration ranges of biological thiols with distinct modes of fluorescence signals[J]. Chemical Science,2016,7(3):1896-1903.