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用于检测细胞中亚硝酰氢的近红外荧光探针及其生物应用
Alternative TitleA Near-Infrared Fluorescent Probe for Detection of Nitroxyl in Living Cells
刘萍; 韩潇玥; 于法标; 陈令新; 于法标,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: fbyu@yic.ac.cn; 陈令新,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: lxchen@yic.ac.cn
2015-12-15
Source Publication分析化学
ISSN0253-3820
Volume43Issue:12Pages:1829-1836
Contribution Rank中国科学院海岸带环境过程与生态修复重点实验室;山东省海岸带环境工程技术研究中心;中国科学院烟台海岸带研究所;中国科学院大学;
Abstract亚硝酰氢(HNO)是NO的单电子质子化还原产物。在治疗心血管疾病中,HNO与NO起不同的药理活性。针对HNO活性高,难以实时检测的问题,本实验设计合成了一种新型不含金属离子的近红外荧光探针ER-JN,用于实时检测模拟生理条件下和活细胞内HNO浓度的变化。此探针由荧光团氮杂氟硼吡咯(AzaBODIPY)和HNO的识别基团三苯基膦两部分组成。探针采用硅胶柱层析分离,洗脱剂为二氯甲烷。产品为墨绿色晶体,产率为28%。此探针具有灵敏性高、选择性强、细胞毒性低等优点,可用于定量检测模拟生理条件下HNO的浓度。检测前后探针的量子产量从0.01增加到0.35。线性范围为0~50μmol/L,探针的检出限(S...; Nitroxyl(HNO),the one-electron reduced and protonated congener of nitric oxide(NO),has been demonstrated with excellent bio-pharmacological effects in cardiovascular disorder treatment,which is distinctive from that of NO.The high reactivity of HNO challenges the accurate detection.To resolve this problem,a near-infrared(NIR) metal-free fluorescent probe,ER-JN,was developed for the detection of intracellular HNO concentration in simulated physiological conditions and living cells.The probe was consisted of two moieties,the A BF2-ehelated tetraarylazadipyrromethane fluorophore(aza-BODIPY) and the HNO recognition unit,diphenylphosphinobenzoyl group.The probe was purified by column chromatography on silica eluting with CH2C12 to give the product as a green solid with a yield of 28%.Our probe exhibited high sensitivity,good selectivity,and low cytotoxic effect,and could be applied to the fluorescent bio-imaging of HNO in simulated physiological conditions.When detected HNO,quantum yield increased from 0.01 to0.35.The linear range located at 0-50 μmol/L.The detection limit was 0.03 μmol/L(S/N = 3).With confocal laser scanning microscope imaging analysis,the probe could detect HNO concentration changes in living cells.Furthermore,the results of flow cytometry confirmed that our probe could be employed for the qualitative and quantitative detection of intracellular HNO concentration level.In this work,we found that probe ER-JN could not only detect HNO in aqueous and in living cells,but also targets endoplasmic reticulum.We anticipated that ER-JN would provide experimental bases in studying physiological and pathological functions of HNO in cells,in vitro and in vivo.
Keyword荧光探针 亚硝酰氢 细胞分析 近红外成像
Department中科院海岸带环境过程与生态修复重点实验室
DOI10.11895/j.issn.0253-3820.150445
Funding Organization国家自然科学基金资助项目(Nos.21405172,21575159,21275158) ; 中国科学院青年创新促进会(Grant2015170)~~
Indexed ByCSCD
Subtype期刊论文
Language中文
CSCD IDCSCD:5591087
Citation statistics
Document Type期刊论文
Identifierhttp://ir.yic.ac.cn/handle/133337/17358
Collection中科院海岸带环境过程与生态修复重点实验室
Corresponding Author于法标,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: fbyu@yic.ac.cn; 陈令新,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: lxchen@yic.ac.cn
Affiliation1.中国科学院海岸带环境过程与生态修复重点实验室
2.山东省海岸带环境工程技术研究中心
3.中国科学院烟台海岸带研究所
4.中国科学院大学
Recommended Citation
GB/T 7714
刘萍,韩潇玥,于法标,等. 用于检测细胞中亚硝酰氢的近红外荧光探针及其生物应用[J]. 分析化学,2015,43(12):1829-1836.
APA 刘萍,韩潇玥,于法标,陈令新,于法标,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: fbyu@yic.ac.cn,&陈令新,中国科学院海岸带环境过程与生态修复重点实验室,E-mail: lxchen@yic.ac.cn.(2015).用于检测细胞中亚硝酰氢的近红外荧光探针及其生物应用.分析化学,43(12),1829-1836.
MLA 刘萍,et al."用于检测细胞中亚硝酰氢的近红外荧光探针及其生物应用".分析化学 43.12(2015):1829-1836.
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