Resveratrol-induced activation of the mitogen-activated protein kinases, ERK1 and ERK2, in human neuroblastoma SH-SY5Y cells

doi:10.1016/S0304-3940(99)00194-9

Neuroscience Letters

Volume 264, Issues 1–3, 2 April 1999, Pages 141-144

https://doi.org/10.1016/S0304-3940(99)00194-9 Get rights and content

Abstract

Phosphorylation of the mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinase 1 (ERK1) and extracellular signal-regulated kinase 2 (ERK2), induced by resveratrol, a natural antioxidant present in grapes and wine, has been studied in vitro on undifferentiated and differentiated (induction by retinoic acid) SH-SY5Y human neuroblastoma cells. In undifferentiated cells resveratrol 1 μM induced phosphorylation of ERK1 and ERK2, which was already evident at 2 min, peaked at 10 min and persisted at 30 min. A wide range (from 1 pM to 10 μM) of resveratrol concentrations were able to induce phosphorylation of ERK1 and ERK2, while higher concentrations (50–100 μM) inhibited MAP kinases phosphorylation. In retinoic acid (RA) differentiated cells resveratrol (1 μM) induced an evident increase in ERK1 and ERK2 phosphorylation. This study demonstrates that resveratrol, even at very low concentrations, may have a biological effect on neuron-like cells.

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Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans-resveratrol
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Citation Excerpt :
This suggests that resveratrol has additional targets linked to the adverse effects on EBs. Among the other potential targets, of particular interest for future studies, are AMPK (Baur et al., 2006; Dasgupta and Milbrandt, 2007), PDE (Park et al., 2012), mechanistic target of rapamycin (mTOR; Liu et al., 2010), and mitogen-activated protein kinase (MAPK; Miloso et al., 1999), as they are key regulators of signaling pathways essential for proper embryo development. One of the health benefit claims of resveratrol is amelioration of diabetes (Szkudelski and Szkudelska, 2015; Öztürk et al., 2017).
Developmental toxicity of compounds, which women of reproductive age are exposed to, should be assessed to minimize the incidence of miscarriage and birth defects. The present study examined the potential developmental toxicity of resveratrol, a dietary supplement widely marketed with various health claims, using the P19C5 embryoid body (EB) morphogenesis assay, which evaluates adverse effects of chemical exposures on tissue growth and axial elongation. Resveratrol (trans isoform) impaired morphogenesis at 4 μM and higher, creating smaller and rounder EBs, whereas cis isoform, and glucuronated and sulfonated metabolites did not. Trans-resveratrol also altered expression levels of developmental regulator genes involved in embryonic patterning, such as Wnt3a, Tbx6, and Cyp26a1. To investigate the mechanisms of trans-resveratrol action, the roles of estrogen receptor, sirtuin 1 (SIRT1), and DNA replication in EB morphogenesis were examined. Neither activators of estrogen receptors (diethylstilbestrol [18 μM] and raloxifene [8 μM]) nor activator of SIRT1 (SRT1720 [2.4–3.2 μM]) caused morphological and molecular alterations that are comparable to trans-resveratrol (10 μM). By contrast, a reduction in the DNA replication rate with aphidicolin (0.4 μM) or hydroxyurea (40 μM) created smaller and rounder EBs and altered the expression levels of Wnt3a, Tbx6, and Cyp26a1 in a manner similar to trans-resveratrol. Consistently, trans-resveratrol significantly reduced the rate of EdU incorporation in P19C5 cells. These results suggest that a reduction in the DNA replication rate is one of the mechanisms by which trans-resveratrol impacts EB development. This study provides mechanistic insight for further investigations on the developmental toxicity of trans-resveratrol.
Oxyresveratrol activates parallel apoptotic and autophagic cell death pathways in neuroblastoma cells
2017, Biochimica et Biophysica Acta - General Subjects
Citation Excerpt :
Here we found that OXYRES was able to activate p38 in both neuroblastoma cells and inactivate ERK1/2, and JNK pathway. When we compare our findings with previous RES experiments, RES upregulated the activity of ERK1 and ERK2 at lower doses but not at higher doses except for p38 MAPK [45] and against pancreatic cancer cells, it downregulated p38 phosphorylation [46]. We showed that pre-treatment with p38 inhibitor SB203580 significantly prevented OXYRES-induced phosphorylation of p38, and autophagy and apoptosis, indicating that OXYRES-induced autophagy and apoptosis is dependent on a p38-activated MAPK cascade.
Drug resistance from apoptosis is a challenging issue with different cancer types, and there is an interest in identifying other means of inducing cytotoxicity. Here, treatment of neuroblastoma cells with oxyresveratrol (OXYRES), a natural antioxidant, led to dose-dependent cell death and increased autophagic flux along with activation of caspase-dependent apoptosis.
For cell viability, we performed the CCK-8 assay. Protein expression changes were with Western blot and immunocytochemistry. Silencing of proteins was with siRNA. The readouts for cell cycle, mitochondria membrane potential, caspase-3, autophagy and apoptosis were performed with flow cytometry.
Phosphorylation of p38 MAPK increased with OXYRES treatment and inhibition of p38 reduced autophagy and cell death from OXYRES. In contrast, PI3K/AKT/mTOR signaling decreased in the target cells with OXYRES and inhibition of PI3K or mTOR enhanced OXYRES-mediated cytotoxicity with increased levels of autophagy. Modulation of either of the apoptosis and autophagy flux pathways affected the extent of cell death by OXYRES, but did not affect the indicators of these pathways with respect to each other. Both pathways were independent of ROS generation or p53 activation.
OXYRES led to cell death from autophagy, which was independent of apoptosis induction. The OXYRES effects were due to changes in the activity levels of p38 MAPK and PI3K/AKT/mTOR.
With two independent and parallel pathways for cytotoxicity induction in target cells, this study puts forward a potential utility for OXYRES or the pathways it represents as novel means of inducing cell death in neuroblastoma cells.
Resveratrol: Multiple Activities on the Biological Functionality of the Cell
2016, Nutraceuticals: Efficacy, Safety and Toxicity
Resveratrol (RV) is a polyphenol nonflavonoid compound present in strongly pigmented vegetables and fruits. This substance has diverse biological activities, including antitumor, antioxidant, antiviral, and phytoestrogenic. We demonstrated that RV inhibits the replication of murine polyomavirus acting during the phase of virion penetration rather than at the nuclear level. At higher concentrations, RV shows also a cytotoxic effect. This complex dose-dependent behavior is not typical of RV; as a matter of fact, literature data show that other natural substances behave in a similar way. Data from our laboratory suggest that the dose-dependent activity of RV is ascribable to alterations of permeability and fluidity of the cell membrane. Here we review data on its antiviral and cytotoxic effects as well as on the cell survival, cycle, and alterations of the membrane parameters. These studies were conducted by “standard” biomolecular/cellular and biophysical strategies such as cytofluorimetry and electrorotation, demonstrating that the main target of RV is the cell membrane. It is suggested that RV may also control cell proliferation. However, further studies are necessary to validate the clinical application of this compound for treatment in antiviral and/or antiproliferative diseases.
The molecular targets of resveratrol
2015, Biochimica et Biophysica Acta - Molecular Basis of Disease
Citation Excerpt :
As with other effectors, it seems that the dosage of Rsv is key to determine MAPK activity. For example, doses up to the low micromolar range might increase ERK signaling, whereas Rsv is inhibitory at the most commonly used, higher, doses (> 50 μM) [133]. This clearly suggests that the impact on this path might be indirect, where MAPKs might act as integrators of diverse signaling paths triggered by Rsv.
Resveratrol has emerged in recent years as a compound conferring strong protection against metabolic, cardiovascular and other age-related complications, including neurodegeneration and cancer. This has generated the notion that resveratrol treatment acts as a calorie-restriction mimetic, based on the many overlapping health benefits observed upon both interventions in diverse organisms, including yeast, worms, flies and rodents. Though studied for over a decade, the molecular mechanisms governing the therapeutic properties of resveratrol still remain elusive. Elucidating how resveratrol exerts its effects would provide not only new insights in its fundamental biological actions but also new avenues for the design and development of more potent drugs to efficiently manage metabolic disorders. In this review we will cover the most recent advances in the field, with special focus on the metabolic actions of resveratrol and the potential role of SIRT1 and AMPK. This article is part of a Special Issue entitled: Resveratrol: Challenges in translating pre-clinical findings to improved patient outcomes.
Inhibition of STIM1 phosphorylation underlies resveratrol-induced inhibition of store-operated calcium entry
2013, Biochemical Pharmacology
Resveratrol, a natural phytoalexin that shows health-promoting benefits, is an inhibitor of store-operated calcium entry (SOCE). Knowledge of the molecular mechanism underlying this inhibition is required for the proper design of therapies that include resveratrol or related stilbenoids, but remains largely unknown. To unravel this mechanism, using HEK293 cells as a model, we found that resveratrol inhibited the ERK1/2 activation triggered by Ca²⁺ store depletion. As a consequence, resveratrol inhibited STIM1 phosphorylation at residues Ser575, Ser608, and Ser621. Because this phosphorylation regulates the dissociation of STIM1 from the microtubule plus-end binding protein EB1 under store depletion conditions, resveratrol inhibited STIM1-EB1 dissociation. This inhibition had downstream effects such as inhibition of STIM1 multimerization in response to store depletion, and a significant impairment in the binding of STIM1 to ORAI1. Although additional targets for resveratrol in the molecular mechanism that governs SOCE cannot be discarded, the present results demonstrate that ERK1/2 pathway is a major target for resveratrol, and that the impairment of its activation produces a significant inhibition of SOCE.
Phenolic Antioxidants as Therapeutic Agents in Major Neurodegenerative Disorders
2022, Health Benefits of Phenolic Antioxidants

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Neuroscience Letters

Abstract

References (20)

Insulin-like Growth Factor-I-mediated neurite outgrowth in vitro requires Mitogen-activated Protein Kinase activation

J. Biol. Chem.

A kinase to remember: dual roles for MAP kinase in long-term memory

Neuron

SHC and GRB-2 are constitutively activated by an epidermal growth factor receptor with a point mutation in the transmembrane domain

J. Biol. Chem.

Wine, alcohol, platelets, and the French paradox for coronary heart disease

Lancet

Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells

J. Biol. Chem.

The MAPK cascade is required for mammalian associative learning

Nat. Neurosci.

Modulatory effect of resveratrol, a natural phytoalexin, on endothelial adhesion molecules and intracellular signal transduction

Pharm. Biol.

Kinetics of trans- and cis-resveratrol (3,4′, 5-trihydroxystilbene) after red wine oral administration in rats

Int. J. Clin. Pharm. Res.

Amelioration of oxidative stress by antioxidants and resveratrol in PC12 cells

NeuroReport

Phenotypic diversification in human neuroblastoma cells: expression of distinct neural crest lineages

Cancer Res.

Cited by (88)

Embryoid body test with morphological and molecular endpoints implicates potential developmental toxicity of trans-resveratrol

Oxyresveratrol activates parallel apoptotic and autophagic cell death pathways in neuroblastoma cells

Resveratrol: Multiple Activities on the Biological Functionality of the Cell

The molecular targets of resveratrol

Inhibition of STIM1 phosphorylation underlies resveratrol-induced inhibition of store-operated calcium entry

Phenolic Antioxidants as Therapeutic Agents in Major Neurodegenerative Disorders