Copper diethyldithiocarbamate as an activator of Nrf2 in cultured vascular endothelial cells.
Copper diethyldithiocarbamate as an activator of Nrf2 in cultured vascular endothelial cells.
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DOI：
10.1007/s00775-016-1337-z
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发表时间：
2016-04
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影响因子：
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通讯作者：
Kaji T
中科院分区：
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作者：
Fujie T;Murakami M;Yoshida E;Tachinami T;Shinkai Y;Fujiwara Y;Yamamoto C;Kumagai Y;Naka H;Kaji T
有机-无机杂化分子作为生物体系分子探针的研究正在迅速发展。这种杂化分子表现出独特的生物活性。其中，二乙基二硫代氨基甲酸铜(Cu10)可激活转录因子NRF2，NRF2负责调节抗氧化剂和II相异源酶。铜络合物在细胞内迅速积累，并诱导Nrf2的核转位，导致下游蛋白表达上调，而没有细胞毒性作用。然而，当二(2-羟乙基)二硫代氨基甲酸铜激活NRF2时，铜离子、二乙基二硫代氨基甲酸酯配体加或不加锌或铁都不能表现出这种活性。Cu10的胞内积累量高于铜(II)和铜(I)。小干扰RNA(SiRNA)介导的铜转运体CTR1基因敲除可减少铜(II)双(二甲基二硫代氨基甲酸)的蓄积，但不影响Cu10的蓄积，表明Cu10通过CTR1非依赖的机制快速进入血管内皮细胞。此外，铜和铁与其他配体的络合物不能激活Nrf2，这表明铜和二硫代氨基甲酸配体之间的分子内相互作用对转录因子的激活是重要的。Cu10诱导NRF2下游蛋白血红素加氧酶-1、NAD(P)H氧化还原酶1和γ-谷氨酰半胱氨酸合成酶的表达。提示Cu10对Nrf2的激活是通过抑制蛋白酶体和结合Kelch样ECH相关蛋白1来实现的。由于Cu10对血管内皮细胞的作用是独特的和多样的，因此铜配合物可能是分析细胞功能的一个很好的分子探针。本文的在线版本(doi：10.1007/s00775-0161337-z)包含补充材料，授权用户可以使用。
The interest in organic–inorganic hybrid molecules as molecular probes for biological systems has been growing rapidly. Such hybrid molecules exhibit unique biological activities. Herein, copper(II) bis(diethyldithiocarbamate) (Cu10) was found to activate the transcription factor NF-E2-related factor 2 (Nrf2), which is responsible for regulating antioxidant and phase II xenobiotic enzymes, in vascular endothelial cells. The copper complex rapidly accumulated within cells and induced nuclear translocation of Nrf2, leading to upregulation of the expression of downstream proteins without cytotoxic effects. However, while copper bis(2-hydroxyethyl)dithiocarbamate activated Nrf2, copper ion, diethyldithiocarbamate ligand with or without zinc or iron failed to exhibit this activity. Intracellular accumulation of Cu10 was higher than that of Cu(II) and Cu(I). While the accumulation of copper(II) bis(dimethyldithiocarbamate) was reduced by small interfering RNA (siRNA)-mediated knockdown of the copper transporter CTR1, the knockdown did not affect Cu10 accumulation, indicating that Cu10 rapidly enters vascular endothelial cells via CTR1-independent mechanisms. In addition, copper and iron complexes with other ligands tested could not activate Nrf2, suggesting that the intramolecular interaction between copper and dithiocarbamate ligand is important for the activation of the transcription factor. Cu10 induced the expression of heme oxygenase-1, NAD(P)H quinone oxidoreductase 1, and γ-glutamylcysteine synthetase, downstream proteins of Nrf2. It was suggested that Cu10-induced activation of Nrf2 was due to proteasome inhibition as well as binding to Kelch-like ECH-associated protein 1. Since the effects of Cu10 on vascular endothelial cells are unique and diverse, the copper complex may be a good molecular probe to analyze the functions of the cells. The online version of this article (doi:10.1007/s00775-016-1337-z) contains supplementary material, which is available to authorized users.