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荧光标记锰配合物的神经保护作用:神经母细胞瘤细胞中 ROS 产生、线粒体膜电位的测定和共聚焦荧光显微镜研究。

Neuroprotective effects of fluorophore-labelled manganese complexes: Determination of ROS production, mitochondrial membrane potential and confocal fluorescence microscopy studies in neuroblastoma cells.

机构信息

Departamento de Química Inorgánica, Facultade de Ciencias, Campus Terra, Universidade de Santiago de Compostela, Lugo, Spain.

Departamento de Farmacología, Facultade de Veterinaria, Campus Terra, Universidade de Santiago de Compostela, Lugo, Spain.

出版信息

J Inorg Biochem. 2022 Feb;227:111670. doi: 10.1016/j.jinorgbio.2021.111670. Epub 2021 Nov 24.

Abstract

In this work, four manganese(II) complexes derived from the ligands HL-HL, that incorporate dansyl or tosyl fluorescent dyes, have been investigated in term of their antioxidant properties. Two of the manganese(II) complexes have been newly prepared using the asymmetric half-salen ligand HL and the thiosemicarbazone ligand HL. The four organic strands and the manganese complexes have been characterized by different analytical and spectroscopic techniques. The study of the antioxidant behaviour of these two new complexes and other two fluorophore-labelled analogues was tested in SH-SY5Y neuroblastoma cells. These four model complexes 1-4 were found to protect cells from oxidative damage in this human neuronal model, by reducing the release of reactive oxygen species. Complexes 1-4 significantly improved cell survival, with levels between 79.1 ± 0.8% and 130.9 ± 4.1%. Moreover, complexes 3 and 4 were able to restore the mitochondrial membrane potential at 1 μM, with 4 reaching levels higher than 85%, similar to the percentages obtained by the positive control agent cyclosporin A. The incorporation of the fluorescent label in the complexes allowed the study of their ability to enter the human neuroblastoma cells by confocal microscopy.

摘要

在这项工作中,研究了四种锰(II)配合物,这些配合物来自包含丹磺酰基或甲苯磺酰基荧光染料的配体 HL-HL,以评估它们的抗氧化性能。其中两种锰(II)配合物是使用不对称半水杨醛配体 HL 和硫代缩氨基脲配体 HL 新制备的。这四种有机链和锰配合物都通过不同的分析和光谱技术进行了表征。通过在 SH-SY5Y 神经母细胞瘤细胞中测试这两种新配合物和其他两种荧光标记类似物的抗氧化行为,研究了它们的抗氧化行为。这四个模型配合物 1-4 被发现可以通过减少活性氧的释放来保护细胞免受氧化损伤。复合物 1-4 显著提高了细胞存活率,水平在 79.1±0.8%至 130.9±4.1%之间。此外,复合物 3 和 4 能够在 1 μM 时恢复线粒体膜电位,其中 4 达到的水平高于 85%,类似于阳性对照药物环孢菌素 A 的百分比。荧光标记的引入使得可以通过共聚焦显微镜研究它们进入人神经母细胞瘤细胞的能力。

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