靶向髓过氧化酶的双模态探针分子的合成与表征

doi:10.7503/cjcu20170817

摘要/Abstract

摘要：

5-羟色胺(5-HT)对动脉粥样硬化斑块中巨噬细胞分泌的髓过氧化酶(MPO)具有靶向功能, 通过酯化反应将5-HT接枝到两亲型聚合物聚衣康酸接枝聚乙二醇接枝十二胺(PIA-g-PEG-g-DDA)上, 制备了具有靶向功能的两亲型聚合物(5-HT-g-PIA-g-PEG-g-DDA). 采用配体交换法, 将超顺磁性氧化铁纳米粒子(SPION)包覆在该聚合物上制备具有磁性的纳米粒子(5-HT-g-PIA-g-PEG-g-DDA@SPION), 通过静电力结合使其与碳点(CDs)结合, 制备具有荧光成像和核磁造影成像的双模态探针的纳米粒子. 采用动态光散射(DLS)及透射电子显微镜(TEM)分析了其粒径和形貌. 采用荧光光谱仪与振动样品磁强计(VSM)测试证明该纳米粒子具备双模态成像功能. 共聚焦成像(CLSM)的测试结果表明, 该纳米试剂对动脉粥样硬化斑块中的巨噬细胞具有靶向性, MTT法的测试结果表明该探针分子具有良好的生物相容性. 该双模态探针具有靶向性好、分辨率高及使用便捷的特点, 是一种性能优异的双模态检测纳米试剂.

关键词: 荧光成像和核磁造影成像双模态探针, 髓过氧化酶, 5-羟色胺, 纳米粒子, 动脉粥样硬化

Abstract:

5-Hydroxytryptamine(5-HT), a targeting molecule for myeloperoxidase from macrophages, was grafted on to (PIA-g-PEG-g-DDA) to prepare a type of amphiphilic polymers with targeting function(5-HT-g-PIA-g-PEG-g-DDA). Superaramagnetic iron-oxide nanoparticles(SPIONs) and fluorescent C dots(CDs) were decorated to the polymers for double-modal imaging. The size and morphology of the obtained nanoprobe were determined. Fluorescence spectrophotometer and vibrating sample magnetometer(VSM) were used to confirm the imaging capability of these bimodal nanoparticles. Confocal laser scanning microscope(CLSM) showed that the nano reagent could target macrophages in atherosclerotic plaque effectively and good biocompatibility was verified by 3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazoliumromide(MTT). The results suggested that the double-modal nano reagent has good targeting ability, high resolution, convenience and is able to become an effective probe for atherosclerosis.

Key words: Double model probe of nuclear magnetic resonance imaging and fluorescent powder imaging, Myeloperoxidase, 5-Hydroxytryptamine, Nanoparticles, Atherosclersis

中图分类号:

O631

TrendMD:

刘晔, 姚顺雨, 方超, 赵外欧, 王静媛, 李亚鹏. 靶向髓过氧化酶的双模态探针分子的合成与表征. 高等学校化学学报, 2018, 39(7): 1573.

LIU Ye, YAO Shunyu, FANG Chao, ZHAO Waiou, WANG Jingyuan, LI Yapeng. Synthesis and Characteristic of Double Model Molecule Probe Targeting Myeloperoxidase. Chem. J. Chinese Universities, 2018, 39(7): 1573.

图/表 11

Scheme 1 Synthesis of CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION

Fig.1 1H NMR spectra of PIA-g-PEG-g-DDA(A)and 5-HT-g-PIA-g-PEG-g-DDA(B)

Fig.2 TEM image of initial 5-HT-g-PIA-g-PEG-g-DDA@SPION(A) and CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION(B)^ Inset is the enlargement of (A).

Fig.3 DLS spectra of 5-HT-g-PIA-g-PEG-g-DDA@SPION(A) and CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION(B)

Fig.4 TGA curve of CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION at 10 ℃/min

Fig.5 MRI test for CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION

Fig.6 Red shift(A), the influence of concentration for intensity of fluorescence(B) and 3D fluorescence spectrum(C) of CDs@5-HT-g-PIA-g-PEG-g-DDA^ c(Fe)/(g·mL-1): (A) a. 0.01; b. 0.05; c. 0.10. (B) a. 0.2 ; b. 0.3; c. 0.4; d. 0.5.

Fig.7 Characteristic of CDs@5-HT-g-PIA-g-PEG-g-DDA fluorescence in macro under 365 nm UV lamp right(A) and under day light(B)

Fig.8 VSM analysis of SPION(A) and CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION(B)

Fig.9 Ccytotoxicity test of CDs@5-HT-PIA-g-PEG-g-DDA@SPION

Fig.10 Cell uptake test of CDs@5-HT-g-PIA-g-PEG-g-DDA@SPION^(A1—A4) were for cell of RAW264.7 under probe; (B1—B4) were free the probe in the cell of RAW264.7; (C1—C4) were for contrast of the endotheliocyte under probe; (D1—D4) were free the probe in the endotheliocyte. (A1—D1) DAPI; (A2—D2) FACIN; (A3—D3) polmer; (A4—D4) MERGE.

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