Novel Targets of Sulforaphane in Primary Cardiomyocytes Identified by Proteomic Analysis
Novel Targets of Sulforaphane in Primary Cardiomyocytes Identified by Proteomic Analysis
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DOI：
10.1371/journal.pone.0083283
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发表时间：
2013-12-11
期刊：
影响因子：
3.7
通讯作者：
Hrelia, Silvana
中科院分区：
文献类型：
--
作者：
Angeloni, Cristina;Turroni, Silvia;Hrelia, Silvana
心血管疾病是工业化世界的主要死亡原因，确定有效的预防战略至关重要。萝卜硫素是一种来自十字花科蔬菜的异硫氰酸酯，已被证明可以上调心肌细胞中的II相酶，并抵消氧化应激诱导的细胞凋亡。本研究的目的是应用蛋白质组学方法鉴定和表征心肌细胞中的萝卜硫素新靶点。二维凝胶电泳和质谱分析用于产生用5 μ M莱菔硫烷处理和未处理1-48小时的原代新生大鼠心肌细胞的蛋白质谱。根据图像分析，发现64个蛋白质点的差异表达和功能的相关性进行了研究，使用MetaCore程序。我们主要集中在3个蛋白：巨噬细胞移动抑制因子（MIF），CLP 36或Elfin，和glycoproteinase 1，由于它们可能参与心脏保护。这些蛋白质的时间依赖性差异表达的验证通过蛋白质印迹法进行。特别是，为了深入了解萝卜硫素对谷胱甘肽酶1的调节的心脏保护作用，使用甲基乙二醛模拟应激进行了进一步的实验。萝卜硫素能够抵消甲基甘氨酸诱导的细胞凋亡，ROS的产生，和应激，可能通过谷胱甘肽酶1上调。在这项研究中，我们首次报道了萝卜硫素的新分子靶点，如MIF，CLP 36和glycoproteinase 1。特别是，我们对萝卜硫素在心肌细胞中的抗心律失常作用提供了新的见解，证实了其在对抗心血管疾病方面的多效性行为。
Cardiovascular diseases represent the main cause of mortality in the industrialized world and the identification of effective preventive strategies is of fundamental importance. Sulforaphane, an isothiocyanate from cruciferous vegetables, has been shown to up-regulate phase II enzymes in cardiomyocytes and counteract oxidative stress-induced apoptosis. Aim of the present study was the identification and characterization of novel sulforaphane targets in cardiomyocytes applying a proteomic approach. Two-dimensional gel electrophoresis and mass spectrometry were used to generate protein profiles of primary neonatal rat cardiomyocytes treated and untreated with 5 mu M sulforaphane for 1-48 h. According to image analysis, 64 protein spots were found as differentially expressed and their functional correlations were investigated using the MetaCore program. We mainly focused on 3 proteins: macrophage migration inhibitory factor (MIF), CLP36 or Elfin, and glyoxalase 1, due to their possible involvement in cardioprotection. Validation of the time-dependent differential expression of these proteins was performed by western blotting. In particular, to gain insight into the cardioprotective role of the modulation of glyoxalase 1 by sulforaphane, further experiments were performed using methylglyoxal to mimic glycative stress. Sulforaphane was able to counteract methylglyoxal-induced apoptosis, ROS production, and glycative stress, likely through glyoxalase 1 up-regulation. In this study, we reported for the first time new molecular targets of sulforaphane, such as MIF, CLP36 and glyoxalase 1. In particular, we gave new insights into the anti-glycative role of sulforaphane in cardiomyocytes, confirming its pleiotropic behavior in counteracting cardiovascular diseases.