Oxygen and Glucose Levels in Cell Culture Media Determine Resveratrol’s Effects on Growth, Hydrogen Peroxide Production, and Mitochondrial Dynamics

Abstract

Resveratrol is a plant-derived polyphenol that has been widely studied for its putative health promoting effects. Many of those studies have been conducted in cell culture, in supra-physiological levels of oxygen and glucose. Resveratrol interacts with reactive oxygen species (ROS) as an antioxidant or pro-oxidant. Resveratrol affects the expression and activities of ROS-producing enzymes and organelles. It is therefore important to consider how cell culture conditions might determine the effects of resveratrol on cultured cells. We determined the effects of resveratrol on cell growth, hydrogen peroxide production, and mitochondrial network characteristics in C2C12 mouse myoblasts and PC3 human prostate cancer cells under conditions of physiological (5%) and supra-physiological (18%) oxygen, and normo- (5 mM) and hyper-glycemia (25 mM). Interestingly, most effects of resveratrol on the parameters measured here were dependent upon prevailing oxygen and glucose levels during the experiment. Many of the effects of resveratrol on cell growth, hydrogen peroxide production, and mitochondrial network characteristics that were seen in 25 mM glucose and/or 18% oxygen were absent under the physiologically relevant conditions of 5 mM glucose with 5% oxygen. These findings emphasize the importance of using physiologically meaningful starting conditions for cell-culture experiments with resveratrol and indeed any manipulation affecting ROS metabolism and mitochondria.

Pure Trans-Resveratrol Nanoparticles Prepared by a Supercritical Antisolvent Process Using Alcohol and Dichloromethane Mixtures: Effect of Particle Size on Dissolution and Bioavailability in Rats

“…Trans-resveratrol, a phytochemical with strong antioxidant property, is naturally found in grapes, berries, and peanuts. The benefits of trans-resveratrol have been reported in various diseases, including cancer, cardiovascular diseases, diabetes mellitus, neurological diseases, obesity, and other conditions associated with oxidative stress and inflammation [1][2][3][4][5]. Chemically, trans-resveratrol is converted into cis-resveratrol by light and has a very low aqueous solubility [6,7].…”

Resveratrol cytotoxicity is energy‐dependent

“…This hypothesis is preliminarily supported by the evidence that in S. cerevisiae at low cellular energy status (0.1% of glucose), 10 μM of resveratrol diminished the cell viability (Madrigal‐Perez et al, ) and that at 0.5% of glucose, the same resveratrol concentration increased H 2 O 2 release (Ramos‐Gomez et al, ), which could enhance the oxidative damage in cells. Additionally, it has been recently reported that 10 μM of resveratrol decreases cell growth in mouse myoblast (C2C12) and human prostate cancer (PC3) under high oxygen conditions (18% of O 2 ) and a low concentration of glucose (5 mM), and this phenotype was accompanied by an increase in H 2 O 2 release rate (Fonseca et al, ). A comparison of two doses of resveratrol (0.00007% and 0.0143%) in Apc Min mice (colorectal carcinogenesis model) fed a high‐fat diet showed that a lower dose of resveratrol suppressed intestinal adenoma development more efficiently than a higher dose (Cai et al, ).…”

“…Indeed, the effects of resveratrol with the mutant HIF‐1α at high glucose are similar to the effects observed with resveratrol and galactose (which only generates ATP through OXPHOS) (Fonseca et al, ), which suggests a possible association among mitochondrial respiration, resveratrol, and HIF‐1α activity. In addition, it has been reported that the resveratrol (10 µM) effects on hydrogen peroxide release, cell growth and the mitochondrial network in C2C12 mouse myoblasts and PC3 human prostate cancer cells are dependent on oxygen levels (5% and 18%) and glucose concentrations (5 and 25 mM) (Fonseca, Moradi, Valente, & Stuart, ). Although the association between resveratrol and HIF‐1α to exert toxic effects is not clear, a possible relationship with the glucose concentration is evident.…”

Natural Compounds in Prostate Cancer Prevention and Treatment: Mechanisms of Action and Molecular Targets

“…This article provides an overview of recent findings about the anti-PCa activity of different natural compounds (Figure 2, Table 1). [47][48][49][50][51][52][53][54]80,100,107,123,124,[135][136][137][138][139]161,163,164,186,190,191,212] The use of phytochemicals for PCa management offers several advantages. Firstly, natural products are safe and well tolerated, as well as usually economically affordable.…”

“…Indeed, treatments aimed at blocking these pathways and/or shifting lactic acid fermentation towards mitochondrial oxidative phosphorylation have shown promise in reducing tumor growth [189]. In recent studies by Fonseca J et al, resveratrol was found to promote a shift towards mitochondrial oxidation in PCa cells concomitantly with the suppression of proliferation, and when this change was prevented by culturing tumor cells in glucose-free medium or via prolyl hydroxylase (PHD) inhibition-mediated stabilization of HIF-1α, the phenol did not affect oxidative phosphorylation and cell growth, indicating that the metabolic shift from glucose fermentation to oxidation is fundamental for its anti-cancer effects [190,191].…”