Guzmán-Pérez Valentina, Bumke-Vogt Christiane, Schreiner Monika, Mewis Inga, Borchert Andrea, Pfeiffer Andreas F H
Department of Clinical Nutrition, German Institute of Human Nutrition, Potsdam-Rehbrücke, Nuthetal, Germany.
Department of Nutrition and Biochemistry, Sciences Faculty-Pontificia Universidad Javeriana, Bogotá D.C, Colombia.
PLoS One. 2016 Sep 13;11(9):e0162397. doi: 10.1371/journal.pone.0162397. eCollection 2016.
Nasturtium (Tropaeolum majus L.) contains high concentrations of benzylglcosinolate. We found that a hydrolysis product of benzyl glucosinolate-the benzyl isothiocyanate (BITC)-modulates the intracellular localization of the transcription factor Forkhead box O 1 (FOXO1). FoxO transcription factors can antagonize insulin effects and trigger a variety of cellular processes involved in tumor suppression, longevity, development and metabolism. The current study evaluated the ability of BITC-extracted as intact glucosinolate from nasturtium and hydrolyzed with myrosinase-to modulate i) the insulin-signaling pathway, ii) the intracellular localization of FOXO1 and, iii) the expression of proteins involved in gluconeogenesis, antioxidant response and detoxification. Stably transfected human osteosarcoma cells (U-2 OS) with constitutive expression of FOXO1 protein labeled with GFP (green fluorescent protein) were used to evaluate the effect of BITC on FOXO1. Human hepatoma HepG2 cell cultures were selected to evaluate the effect on gluconeogenic, antioxidant and detoxification genes and protein expression. BITC reduced the phosphorylation of protein kinase B (AKT/PKB) and FOXO1; promoted FOXO1 translocation from cytoplasm into the nucleus antagonizing the insulin effect; was able to down-regulate the gene and protein expression of gluconeogenic enzymes; and induced the gene expression of antioxidant and detoxification enzymes. Knockdown analyses with specific siRNAs showed that the expression of gluconeogenic genes was dependent on nuclear factor (erythroid derived)-like2 (NRF2) and independent of FOXO1, AKT and NAD-dependent deacetylase sirtuin-1 (SIRT1). The current study provides evidence that BITC might have a role in type 2 diabetes T2D by reducing hepatic glucose production and increasing antioxidant resistance.
旱金莲(Tropaeolum majus L.)含有高浓度的苄基葡萄糖硫苷。我们发现苄基葡萄糖硫苷的一种水解产物——苄基异硫氰酸酯(BITC)——可调节转录因子叉头框O1(FOXO1)的细胞内定位。FoxO转录因子可拮抗胰岛素作用,并触发参与肿瘤抑制、长寿、发育和代谢的多种细胞过程。本研究评估了从旱金莲中提取为完整葡萄糖硫苷并用黑芥子酶水解的BITC调节以下方面的能力:i)胰岛素信号通路,ii)FOXO1的细胞内定位,以及iii)参与糖异生、抗氧化反应和解毒的蛋白质的表达。使用稳定转染了用绿色荧光蛋白(GFP)标记的FOXO1蛋白的组成型表达的人骨肉瘤细胞(U-2 OS)来评估BITC对FOXO1的影响。选择人肝癌HepG2细胞培养物来评估对糖异生、抗氧化和解毒基因及蛋白质表达的影响。BITC降低了蛋白激酶B(AKT/PKB)和FOXO1的磷酸化;促进FOXO1从细胞质向细胞核的转运,拮抗胰岛素作用;能够下调糖异生酶的基因和蛋白质表达;并诱导抗氧化和解毒酶的基因表达。用特异性小干扰RNA(siRNA)进行的敲低分析表明,糖异生基因的表达依赖于核因子(红系衍生)样2(NRF2),且独立于FOXO1、AKT和烟酰胺腺嘌呤二核苷酸(NAD)依赖性脱乙酰酶沉默调节蛋白1(SIRT1)。本研究提供了证据表明,BITC可能通过减少肝脏葡萄糖生成和增加抗氧化抗性在2型糖尿病(T2D)中发挥作用。