荧光多肽探针靶向检测细胞和斑马鱼中的钆离子  

A Novel Fluorescent Peptide Probe for Specific Detection of Gadolinium Ion(Ⅲ)in Cells and Zebrafish

作　　者：赵闯 李佳楠 孙秀霞 肖建喜[1] ZHAO Chuang;LI Jia-Nan;SUN Xiu-Xia;XIAO Jian-Xi(College of Chemistry and Chemical Engineering,Lanzhou University,Lanzhou 730000,China;School of Pharmacy,Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学化学化工学院,兰州730000 [2]兰州大学药学院,兰州730000

出　　处：《分析化学》2022年第7期1014-1021,I0001-I0003,共11页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.22074057,21775059)资助。

摘　　要：含钆造影剂作为临床应用最广泛的磁共振造影剂,在人体疾病的临床诊断和相关研究中发挥着非常重要的作用,而生物体内钆沉积被发现与肾源性系统性纤维化密切相关,并可能是多发性硬化、神经纤维瘤病、遗传性代谢紊乱等疾病的关键诱因。因此,实现生物体内游离的钆离子(Gd^(3+))高效检测对相关疾病的机理研究至关重要。本研究制备了一种新型荧光多肽探针PT(TPE-Glu-Glu-Glu-Pro-Gly-Glu-Glu-Glu),利用1-(4-羧基苯)-1,2,2-三苯乙烯(TPE)修饰的多肽探针PT,实现了溶液和体液中Gd^(3+)的高效检测,并且不受其它常见金属离子的干扰;同时,Job工作曲线证明,荧光多肽探针PT与Gd^(3+)以1∶3的方式结合,结合常数为2.34×10^(15)L^(3)/mol^(3)。在HEPES缓冲液和唾液样品中,随着Gd^(3+)的加入,荧光多肽探针PT呈现“Turn-On”型响应模式,检出限分别为39和53 nmol/L,并成功用于小鼠成纤维细胞(L929细胞)和斑马鱼中Gd^(3+)的靶向成像。此荧光多肽探针PT具有水溶性好、细胞毒性低等特性,可用于斑马鱼和L929细胞中Gd^(3+)的高灵敏检测,在人体内残留Gd^(3+)的分子诊断方面具有巨大的应用潜力。As the most widely used magnetic resonance imaging contrast in clinical applications,gadolinium-based contrast agents play an important role in the clinical diagnosis and related research,while gadolinium deposition has been found to be closely related to nephrogenic systemic fibrosis and may cause multiple sclerosis,neurofibromatosis,inborn error of metabolism and other diseases.Therefore,the development of efficient assays for free Gd^(3+) is critical for the mechanistic study of these diseases.In this work,the fluorescent peptide probe PT consisted of(TPE-Glu-Glu-Glu-Pro-Gly-Glu-Glu-Glu)sequences was reported.The results showed that the efficient detection of Gd^(3+) in solution and biological fluids using the TPE-modified peptide probe PT without interferences of other metal ions was achieved.Meanwhile,the Job′s analysis result indicated that the probe PT and Gd^(3+) formed a 1∶3 complex and recovered its fluorescence,with a binding constant of 2.34×10^(15)L^(3)/mol^(3).It was demonstrated that the fluorescent peptide probe PT provided a“Turn-On”sensing mode for Gd^(3+) with detection limits of 39 and 53 nmol/L in HEPES buffer solution and saliva samples.The probe PT was successfully used for imaging of Gd^(3+) in L929 cells and zebrafish.Notably,the newly developed fluorescent peptide probe PT had extraordinary features such as highly water solubility,low biotoxicity and high specificity for Gd^(3+) in zebrafish and L929 cells,which showed great potential in the molecular diagnosis of the concentration of residual Gd^(3+) in human body.

关 键 词：荧光多肽探针 钆离子 L929细胞 斑马鱼 

分 类 号：O657.3[理学—分析化学] R965[理学—化学]