Resveratrol promotes cellular glucose utilization in primary cultured cortical neurons via calcium-dependent signaling pathway

doi:10.1016/j.jnutbio.2012.02.015

The Journal of Nutritional Biochemistry

Volume 24, Issue 4, April 2013, Pages 629-637

https://doi.org/10.1016/j.jnutbio.2012.02.015 Get rights and content

Abstract

Background and Purpose

Impairment of glucose utilization contributes to neuronal degeneration of Alzheimer's disease patients. Cellular glucose utilization can be regulated by calcium-dependent signaling pathways. Resveratrol (RSV) is a plant-derived polyphenol with multiple beneficial effects, including neuroprotection and metabolic improvement. Here, we investigated the effect of RSV on neuronal calcium signal and glucose utilization.

Experimental Methods

Primary culture of cortical neurons, calcium imaging, 2-NBDG assay and western blotting were employed to investigate RSV-mediated effects on neuronal calcium signal and glucose utilization.

Results

RSV elevated intracellular calcium in cortical neurons via modulation of secondary messenger system including nitrous oxide, cGMP and cAMP. Secondarily, a calcium-dependent enhancement of neuronal glucose utilization after RSV treatment was observed. The effects on neuronal glucose utilization are largely dependent on RSV-induced calcium-dependent AMP-activated protein kinase activation.

Conclusion

Our findings show that activation of calcium-dependent signaling pathways by RSV may convey improvements of neuronal glucose utilization.

Introduction

Neurons are particularly sensitive to fluctuations in energy levels. Energy deficiency is tightly related to neuronal survival and viability and also contributes to age-related disorders, e.g., Alzheimer's disease (AD) [1], [2]. Glucose utilization provides energy for functional neuronal activities. A family of glucose transporters (GluTs) is believed to be responsible for the majority of glucose utilization in brain [3], [4]. Inhibition of neuronal glucose utilization may decrease neuronal activity and precede neuronal degeneration. Restrictions to glucose availability increase the sensitivity of primary neurons to glutamate excitation [5], [6]. The impairment of neuronal glucose utilization in the brain of AD patients may precede neuronal degeneration [7], [8]. Previous studies support the view that nutrients can serve as trophic factors to promote neuronal repair and survival following injury [9]. Thus, nutrients may improve the neurological disorders including AD.

Resveratrol (RSV), a polyphenol widely found in red wine, peanuts and pomegranate, has received tremendous attention over the past couple of decades, due to its benefits such as antioxidation, regulation of energy metabolism, neuroprotection and anti-aging effect [10], [11], [12], [13]. Calorie restriction (CR) exhibits anti-aging effects on many species via metabolic improvements and RSV exerts multiple beneficial effects similar to those associated with CR [14], [15], [16], [17], [18], [19], [20]. Thus, more evidences support the notion that a number of benefits of RSV, such as anti-aging and neuroprotection are attributable to the multiple metabolic improvements [20], [21]. Recently, it has been reported that RSV inhibits Aβ accumulation via calcium-dependent signaling pathway [21]. Calcium is an important second messenger in cellular energy metabolism, involved in the docking and fusion of glucose transporter 4 (GluT4) vesicles with plasma membrane [22], [23]. AMP-activated protein kinase (AMPK), a critical regulator of cellular energy metabolism, is also indirectly activated by the activation of Ca^2 + sensitive calmodulin-dependent protein kinase kinases (CaMKKs) [24], [25]. In particular, AMPK activation results in the increase in cellular glucose uptake [26], [27], suggesting that intracellular calcium signal may regulate neuronal glucose utilization by AMPK activation. Furthermore, a transient elevation of intracellular calcium concentration promotes the vesicle exocytosis and translocation of GluT3. This exocytosis can be inhibited by amyloid beta-peptide [28], [29].

However, until now, it is not fully understood the exact mechanism by which RSV regulates cytosolic calcium levels. Therefore the precise mechanism underlying RSV -mediated calcium signaling was explored in the present study. Moreover, since previous studies show that CR can facilitate cellular glucose utilization [30], [31], it is likely that RSV may produce a mimetic effect of CR on glucose utilization in neurons via calcium-dependent signaling pathways. Thus, in this study, we intended to explore the effect of RSV-triggered intracellular calcium events on neuronal glucose utilization as well as the underlying mechanisms.

Section snippets

Materials

Resveratrol (RSV), thapsigargin (TG), cyclopiazonic acid (CPA), BAPTA-AM, KT5823, dithiothreitol (DTT), Compound C, ICI 182780, BFA, nicotinamide, 7-oxo-7H-benzimidazo[2,1-a]benz[de]isoquinoline-3-carboxylic acid-acetic acid (STO609), NG-nitro-l-arginine methyl ester (L-NAME), 1H-[1,2,4] oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), 5-aminoimidazole-4-carboxamide riboside (AICAR), N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H89), and

RSV increase [Ca^2 +]_i in a dose-dependent manner by mobilization of intracellular calcium stores

In the first set of experiments, we employed the calcium imaging technique to investigate the dynamic alteration of intracellular calcium mediated by RSV in cultured rat prefrontal cortical neurons. The vehicle, 0.1% DMSO, exhibited no effect on the [Ca^2 +]_i of cortical neurons (data not shown). Incubation of cortical neurons with RSV (5–100 μM) increased [Ca^2 +]_i in a dose-dependent manner by 42.30±0.53% over basal levels at 5 μM, 138±1 % at 10 μM, 201±8 % at 50 μM and 365±5% at 100 μM (n= 12,

Discussion

In the current study, we have shown that RSV produces an intracellular calcium elevation in cortical neurons via modulation of secondary messenger system including NO, cGMP and cAMP. Once calcium enters the neuron, a calcium-dependent AMPK activation is initiated. As a result, the intracellular glucose utilization is enhanced in a calcium-dependent way (Fig. 7).

Resveratrol, a polyphenol that is present at high levels in the skin of grapes, nuts and red wine, attracts more and more attention for

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China to Dr. J.G. Chen (No.30930104, No. 81171258), Dr. F. Wang (No. 81173039) and Dr. L.H. Long (No. 30801390).

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¹: Equally contributed authors.

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Research articleResveratrol promotes cellular glucose utilization in primary cultured cortical neurons via calcium-dependent signaling pathway

Abstract

Background and Purpose

Experimental Methods

Results

Conclusion

Introduction

Section snippets

Materials

RSV increase [Ca2 +]i in a dose-dependent manner by mobilization of intracellular calcium stores

Discussion

Acknowledgments

Exp Neurol

J Neurol Sci

J Nutr Biochem

J Biol Chem

J Biol Chem

Curr Opin Pharmacol

Biochem Biophys Res Commun

Exp Neurol

Arch Biochem Biophys

Free Radic Biol Med

Eur J Pharmacol

J Biol Chem

J Nutr

Eur J Pharmacol

Biochem Biophys Res Commun

J Biol Chem

Inherent abnormalities in energy metabolism in Alzheimer disease. Interaction with cerebrovascular compromise

Ann N Y Acad Sci

Brain metabolism and brain disease: is metabolic deficiency the proximate cause of Alzheimer dementia?

J Neurosci Res

Glucose transporter proteins in brain

FASEB J

Glucose transport in brain and retina: implications in the management and complications of diabetes

Diabetes Metab Res Rev

Regulation of glucose transporter 3 surface expression by the AMP-activated protein kinase mediates tolerance to glutamate excitation in neurons

J Neurosci

Decreased brain glucose metabolism in microvessels from patients with Alzheimer's disease

Ann N Y Acad Sci

Effect of antioxidants, resveratrol, quercetin, and N-acetylcysteine, on the functions of cultured rat hepatic stellate cells and Kupffer cells

Hepatology

Neuroprotective abilities of resveratrol and other red wine constituents against nitric oxide-related toxicity in cultured hippocampal neurons

Br J Pharmacol

Resveratrol improves health and survival of mice on a high-calorie diet

Nature

Anti-aging properties of resveratrol: review and report of a potent new antioxidant skin care formulation

J Cosmet Dermatol

Antiaging, longevity and calorie restriction

Curr Opin Clin Nutr Metab Care

Research article
Resveratrol promotes cellular glucose utilization in primary cultured cortical neurons via calcium-dependent signaling pathway

RSV increase [Ca^2 +]_i in a dose-dependent manner by mobilization of intracellular calcium stores