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Production of Reactive Oxygen Species and Changes in Antioxidant Enzyme Activities during Differentiation of 3T3-L1 Adipocyte

Abstract

Obesity, a major public health problem around the world, is a strong risk factor for the development of type 2 diabetes, atherosclerosis, hypertension and cardiovascular diseases. Recent research suggests that increased production of reactive oxygen species (ROS) from accumulated fat in obesity leads to elevated systemic oxidative stress and contributes to the development of obesity-linked chronic disorders. The aim of the current study was to investigate changes of key enzymes associated with antioxidant response and metabolic pathways in 3T3-L1 cell, a preadipocyte cell line that undergoes differentiation into mature adipocytes. The changes in lipid accumulation, ROS production, glucose-6-phosphate dehydrogenase (G6PDH), superoxide dismutase (SOD) and catalase (CAT) during the course of differentiation were determined. The ROS production and G6PDH activity exponentially increased as differentiation progressed. However, CAT activity showed a sharp decrease until day 2, followed by a gradual increase up to day 6 and then leveled off. Given the importance of adipocyte differentiation and ROS production in oxidation-linked diseases, these data provide a biochemical rationale for further studies to identify cellular mechanisms that can counter ROS generation and modulate cellular redox environment in adipocytes.

Abbreviations

AER:

antioxidant enzyme responses

BCS:

bovine calf serum

CAT:

catalase

DETAPAC:

diethylene triaminie penta-acetic acid

DEX:

dexamethasone

DMEM:

dulbecco’s modified Eagle’s medium; ETS, electron transport system

FBS:

fetal bovine serum

G6PDH:

glucose-6-phosphate dehydrogenase

ROS:

reactive oxygen species

HMP:

hexose monophosphate

IBMX:

3-isobutyl-1-methylxanthine

NBT:

nitroblue tetrazolium

NO:

nitric oxide

NOX:

NADPH oxidase

OD:

optical density

PBS:

phosphate buffered saline

P/S:

penicillin-streptomycin

SOD:

superoxide dismutase

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Correspondence to Young-Cheul Kim.

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Lee, OH., Kwon, YI., Hong, HD. et al. Production of Reactive Oxygen Species and Changes in Antioxidant Enzyme Activities during Differentiation of 3T3-L1 Adipocyte. J. Korean Soc. Appl. Biol. Chem. 52, 70–75 (2009). https://doi.org/10.3839/jksabc.2009.012

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  • DOI: https://doi.org/10.3839/jksabc.2009.012

Key words

  • antioxidant enzyme response
  • glucose-6-phosphate dehydrogenase
  • hexose monophosphate shunt
  • reactive oxygen species
  • 3T3-L1 adipocyte differentiation