Sulforaphane (SF), a naturally occurring isothiocyanate abundant in Cruciferous vegetables, has gained attention as a potential chemopreventive compound (1) thanks to its ability to induce several classes of genes implicated in reactive oxygen species (ROS) and electrophiles detoxification. Antioxidant responsive element (ARE)- mediated gene induction is a pivotal mechanism of cellular defence against the toxicity of electrophiles and ROS. The transcription factor NF-E2-related factor-2 (Nrf2), is essential for the up-regulation of these genes. Nrf2 phosphorylation by protein kinases such as Akt kinase, extracellular-signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK) and protein kinase C (PKC) influence the activation of the Nrf2/ARE pathway (2, 3). Recently, we have demonstrated that SF elicits cardioprotective activity by up-regulating a battery of antioxidants and phase II enzymes (4). The aim of this study was to verify SF mechanism of action leading to cardioprotection by investigating the effect of SF treatment on Nrf2 activation in primary cultures of neonatal rat cardiomyocytes. In particular we focused our attention on the ability of SF to activate different mitogen–activated protein kinases (MAPK) related to Nrf2 translocation to the nucleus. Cultured rat cardiomyocytes, prepared and grown as previously reported (5), were supplemented with 5 μM SF for different times. Western blot analyses of Nrf2, phsopho-p38, phospho-JNK, phospho-Akt, phospho-ERK1/2 were performed using specific antibodies and following the manufacturer’s recommended protocols. Nrf2 translocation to the nucleus was evaluated by immunoblotting of different cellular fractions. The expression of Nrf2 was determined by RT-PCR. SF was able to modulate MAPK phosphorylation and Nrf2 translocation to the nucleus. This translocation was blocked by cell treatment with specific MAPK inhibitors demonstrating that Nrf2 activation is mediated by these pathways. SF was also able to up-regulate Nrf2 expression. So, the cardioprotective activity of SF could be ascribed to its ability to modulate both MAPK signaling pathways and Nrf2 expression. This research was supported by MIUR-COFIN and Fondazione del Monte di Bologna e Ravenna 1. Y. Zhang, et al. (1992) Proc Natl Acad Sci U S A. 89, 2399-403. 2. J. D. Hayes, et al. (2001) Cancer Lett. 174, 103-13. 3. T. Nguyen, et al. (2004) Free Radic Biol Med. 37, 433-41. 4. C. Angeloni, et al. (2007) Proc Nutr Soc. 66, 111A. 5. S. Hrelia, et al. (2002) Biochim Biophys Acta. 1567, 150-6.
Sulforaphane modulates NRF2/ARE pathway in cardiac cells / E. Leoncini; C. Angeloni; M. Malaguti; S. Angelini; P. Hrelia; S. Hrelia. - STAMPA. - (2008), pp. 4.17-4.17. (Intervento presentato al convegno 53° Congresso della Società Italiana di Biochimica e Biologia Molecolare. tenutosi a Riccione nel 23-26 Settembre).
Sulforaphane modulates NRF2/ARE pathway in cardiac cells.
LEONCINI, EMANUELA;ANGELONI, CRISTINA;MALAGUTI, MARCO;ANGELINI, SABRINA;HRELIA, PATRIZIA;HRELIA, SILVANA
2008
Abstract
Sulforaphane (SF), a naturally occurring isothiocyanate abundant in Cruciferous vegetables, has gained attention as a potential chemopreventive compound (1) thanks to its ability to induce several classes of genes implicated in reactive oxygen species (ROS) and electrophiles detoxification. Antioxidant responsive element (ARE)- mediated gene induction is a pivotal mechanism of cellular defence against the toxicity of electrophiles and ROS. The transcription factor NF-E2-related factor-2 (Nrf2), is essential for the up-regulation of these genes. Nrf2 phosphorylation by protein kinases such as Akt kinase, extracellular-signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK) and protein kinase C (PKC) influence the activation of the Nrf2/ARE pathway (2, 3). Recently, we have demonstrated that SF elicits cardioprotective activity by up-regulating a battery of antioxidants and phase II enzymes (4). The aim of this study was to verify SF mechanism of action leading to cardioprotection by investigating the effect of SF treatment on Nrf2 activation in primary cultures of neonatal rat cardiomyocytes. In particular we focused our attention on the ability of SF to activate different mitogen–activated protein kinases (MAPK) related to Nrf2 translocation to the nucleus. Cultured rat cardiomyocytes, prepared and grown as previously reported (5), were supplemented with 5 μM SF for different times. Western blot analyses of Nrf2, phsopho-p38, phospho-JNK, phospho-Akt, phospho-ERK1/2 were performed using specific antibodies and following the manufacturer’s recommended protocols. Nrf2 translocation to the nucleus was evaluated by immunoblotting of different cellular fractions. The expression of Nrf2 was determined by RT-PCR. SF was able to modulate MAPK phosphorylation and Nrf2 translocation to the nucleus. This translocation was blocked by cell treatment with specific MAPK inhibitors demonstrating that Nrf2 activation is mediated by these pathways. SF was also able to up-regulate Nrf2 expression. So, the cardioprotective activity of SF could be ascribed to its ability to modulate both MAPK signaling pathways and Nrf2 expression. This research was supported by MIUR-COFIN and Fondazione del Monte di Bologna e Ravenna 1. Y. Zhang, et al. (1992) Proc Natl Acad Sci U S A. 89, 2399-403. 2. J. D. Hayes, et al. (2001) Cancer Lett. 174, 103-13. 3. T. Nguyen, et al. (2004) Free Radic Biol Med. 37, 433-41. 4. C. Angeloni, et al. (2007) Proc Nutr Soc. 66, 111A. 5. S. Hrelia, et al. (2002) Biochim Biophys Acta. 1567, 150-6.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
