Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation
Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation
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DOI：
10.1016/j.atherosclerosis.2017.08.012
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发表时间：
2017-10-01
期刊：
影响因子：
5.3
通讯作者：
Pasch, Andreas
中科院分区：
文献类型：
--
作者：
Aghagolzadeh, Parisa;Radpour, Ramin;Pasch, Andreas
背景和目标：血管钙化是一种常见的健康问题，与氧化应激、炎症和循环钙蛋白颗粒（CPP）有关。硫化氢是一种内源性信号分子，具有抗氧化特性和靶向氧化还原信号的药物开发潜力。然而，其作用于血管平滑肌细胞（VSMC）钙化的分子机制尚未阐明。因此，我们试图确定的钙化抑制特性的硫化物采用我们最近开发的CPP诱导的VSMC calcification model.Methods的关键途径：使用下一代测序，我们调查了硫氢化钠治疗与非治疗钙化VSMCs的转录组学变化。结果：CPP可引起细胞内钙离子的明显积累，硫化物可使其浓度依赖性降低。CPP诱导的过氧化氢产生和增强的促炎/氧化应激相关基因表达特征均被硫化物处理减弱。我们的转录组数据的基因本体富集和计算机途径分析表明，NAD（P）H脱氢酶[醌] 1（NQO 1）作为潜在的调解人。证实了这些发现，沉默Kelch样ECH相关蛋白1（KEAP 1），核因子（红细胞衍生2）样2（NRF 2）核活性的抑制剂，增强NQO 1的表达，而NRF 2沉默减少NQO 1的表达和废除的钙化抑制活性的硫化物。此外，免疫荧光显微镜和Western blot分析证实了NRF 2的核转位的硫化物在VSMC.Conclusions：硫化物衰减CPP诱导的VSMC钙化在体外通过KEAP 1-NRF 2氧化还原传感/应激反应系统，通过增强NQO 1的表达。(C)2017爱思唯尔B. V.保留所有权利。
Background and aims: Vascular calcification is a common health problem related to oxidative stress, inflammation, and circulating calciprotein particles (CPP). Hydrogen sulfide is an endogenous signaling molecule with antioxidant properties and potential for drug development targeting redox signaling. Yet, its molecular mechanisms of action in vascular smooth muscle cell (VSMC) calcification have not been delineated. We therefore sought to identify key pathways involved in the calcification-inhibitory properties of sulfide employing our recently developed CPP-induced VSMC calcification model.Methods: Using next-generation sequencing, we investigated the transcriptomic changes of sodium hydrosulfide-treated versus non-treated calcifying VSMCs. The potential role of candidate genes and/or regulatory pathways in prevention of calcification was investigated by small interfering RNA (siRNA).Results: CPP led to a pronounced accumulation of cell-associated calcium, which was decreased by sulfide in a concentration-dependent manner. Both, CPP-induced hydrogen peroxide production and enhanced pro-inflammatory/oxidative stress-related gene expression signatures were attenuated by sulfide-treatment. Gene ontology enrichment and in silico pathway analysis of our transcriptome data suggested NAD(P) H dehydrogenase [quinone] 1 (NQO1) as potential mediator. Corroborating these findings, silencing of Kelch-like ECH-associated protein 1 (KEAP1), an inhibitor of nuclear factor (erythroid-derived 2)-like 2 (NRF2) nuclear activity, enhanced NQO1 expression, whereas NRF2 silencing reduced the expression of NQO1 and abrogated the calcification-suppressing activity of sulfide. Moreover, immunofluorescence microscopy and Western blot analysis confirmed nuclear translocation of NRF2 by sulfide in VSMC.Conclusions: Sulfide attenuates CPP-induced VSMC calcification in vitro via the KEAP1-NRF2 redox sensing/stress response system by enhancing NQO1 expression. (C) 2017 Elsevier B.V. All rights reserved.