Effects of oxidative stress on adiponectin secretion and lactate production in 3T3-L1 adipocytes

doi:10.1016/j.freeradbiomed.2004.12.010

Free Radical Biology and Medicine

Volume 38, Issue 7, 1 April 2005, Pages 882-889

https://doi.org/10.1016/j.freeradbiomed.2004.12.010 Get rights and content

Abstract

Obesity is an increasing nutritional disorder in developed countries, and oxidative stress has been identified as a key factor in numerous pathologies such as diabetes, inflammation, and atherosclerosis, which are favored by obesity. The objective of the present study was to investigate the effects of oxidative stress in 3T3-L1 adipose cells on two parameters involved in metabolic complications associated with obesity, namely adiponectin secretion and lactate production. Differentiated 3T3-L1 adipose cells were exposed to increasing concentrations of glucose oxidase. 4-Hydroxynonenal (4-HNE), a relevant lipid peroxidation by-product which may affect several metabolic processes in making covalent adducts with various molecules; adiponectin secretion; and lactate production were measured in response to glucose oxidase exposure. Results show an inhibition of adiponectin mRNA expression by glucose oxidase and a significant inverse correlation between 4-HNE formation and adiponectin secretion. Furthermore, 4-HNE alone inhibits adiponectin production by 3T3-L1. On the other hand, glucose oxidase and 4-HNE significantly stimulated lactate production by 3T3-L1 adipocytes. These results demonstrate that adipose cells are highly sensitive to oxidative stress, with subsequent decreased adiponectin secretion and increased lactate production, two events involved in the development of insulin resistance.

Introduction

Obesity is the most common and fastest growing nutritional disorder in industrial and emergent countries. Recent studies have pointed out that obesity is a state of low-grade permanent inflammation [1], [2], [3]. Inflammation is closely linked to oxidative stress [4], which has been found significantly correlated to body mass index [5]. The response of adipocytes to such a stress has never been thoroughly investigated. Some studies have tested the effect induced by oxidative molecules such as hydrogen peroxide on different metabolic events in adipose cells [6], but the mediators produced in response to oxidative stress have rarely been proposed and never quantified in adipose cells.

The hydroxyalkenal 4-hydroxynonenal (4-HNE) is widely recognized as a specific marker of oxidative stress [7], and it has been reported to alter a number of biomolecules in making covalent adducts with them [8]. It derives from peroxidation of n-6 polyunsaturated fatty acids such as arachidonic and linoleic acids, the two most common fatty acids in biomembranes, except in the cerebrovascular system, which makes 4-HNE a relevant marker for oxidative stress.

In the present study, the metabolic effect of oxidative stress was studied on two metabolic functions of adipose cells, adiponectin secretion and lactate production, both parameters playing a part in insulin resistance. Indeed, adiponectin expression has been associated with higher degrees of insulin sensitivity [9], and lactate induces insulin resistance in skeletal muscle [10]. Adiponectin is a protein synthesized exclusively by adipocytes [11], [12], [13], [14]. Recent studies suggest that it plays a critical role in the regulation of insulin sensitivity and exhibits putative antiatherogenic properties [15]. Although adiponectin is secreted only from adipose tissue, plasma adiponectin concentration is decreased in obesity for unknown reasons. Our working hypothesis is that adiponectin production could be sensitive to oxidative stress. On the other hand, lactate production by adipose tissue has been recognized as a novel function of adipose tissue [16].

The objective of the present study was then to submit 3T3-L1 adipocytes to oxidative stress, measure the formation of 4-HNE, and investigate adiponectin secretion and lactate production.

Section snippets

Cell culture

3T3-L1 fibroblasts were obtained from American Type Culture Collection (Manassas, VA, USA). The cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% newborn calf serum, 4 mM glutamine, 4 nM insulin (Actrapid Human; Novo), 10 mM Hepes, 25 μg sodium ascorbate, 100 IU penicillin, 100 μg streptomycin, and 0.25 μg amphotericin B per milliliter, at 37°C in a water-saturated atmosphere with 5% CO₂ in air, in a Heraeus incubator (BB16). Adipocytes were studied 17 days after

Results

Hydrogen peroxide concentrations in 3T3-L1 cell culture significantly increased in response to glucose oxidase concentrations until 50 mU/ml, compared to control (Fig. 1). Glucose concentration in the culture buffer decreased in the presence of glucose oxidase. That diminution was already significant for the smallest glucose oxidase concentration (1 mU/ml). After 16 h incubation with the highest concentrations of glucose oxidase (50 and 100 mU/ml), more than 1.5 mM glucose was still present in

Discussion

For decades, adipocytes have been considered as low-reactive cells. Indeed, it was thought that they were able only to increase or decrease their size according to the overall metabolism, resulting in either lipogenic or lipolytic conditions. More recently, the increased prevalence of obesity in occidentalized populations has raised an important research effort on adipose tissue. Among the most important discoveries, it has been shown that adipocytes produce numerous regulatory factors,

Acknowledgments

This work was supported by INSERM, IMBL, and Région Rhône-Alpes. A.F. Soares is a recipient of CAPES BRAZIL (No. 0159 026) for his Ph.D. preparation.

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Original contributionEffects of oxidative stress on adiponectin secretion and lactate production in 3T3-L1 adipocytes

Abstract

Introduction

Section snippets

Cell culture

Results

Discussion

Acknowledgments

Surgery

Nutrition

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Free Radic. Biol. Med.

J. Lipid Mediat. Cell Signaling

J. Biol. Chem.

Eur. J. Pharmacol.

Biochem. Biophys. Res. Commun.

Free Radic. Biol. Med.

Biochem. Biophys. Res. Commun.

Methods Enzymol.

Am. J. Hypertens

Chem. Phys. Lipids

Am. J. Clin. Nutr.

Minireview: the adipocyte at the crossroad of energy homeostasis, inflammation and atherosclerosis

Endocrinology

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Original contribution
Effects of oxidative stress on adiponectin secretion and lactate production in 3T3-L1 adipocytes