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Applied Biological Chemistry
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  • Food Science/Microbiology
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Production of Reactive Oxygen Species and Changes in Antioxidant Enzyme Activities during Differentiation of 3T3-L1 Adipocyte

Journal of the Korean Society for Applied Biological Chemistry volume 52, pages 70–75 (2009)Cite this article

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

References

  • Araki S, Dobashi K, Kubo K, Yamamoto Y, Asayama K, and Shirahata A (2006) N-acetylsysteine attenuates TNF-? induced changes in secretion of interleukin-6, plasminogen activator inhibitor-1 and adiponectin from 3T3-L1 adipocytes. Life Sci 79, 2405–2412.

    Article  CAS  Google Scholar 

  • Beers R and Sizer IA (1952) Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195, 133–140.

    CAS  Google Scholar 

  • Blumberg J M, Tzameli I, Astapova I, Lam FS, Flier JS, and Hollenberg AN (2006) Complex role of the vitamin D receptor and its ligand in adipogenesis in 3T3-L1 cells. J Biol Chem 28, 11205–11213.

    Article  Google Scholar 

  • Bradford MA (1976) Rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248–254.

    Article  CAS  Google Scholar 

  • Deutsch J (1983) Glucose-6-phosphate dehydrogenase. In Methods of enzymatic analysis (3rd ed), pp. 191–197. Verlag Chemie Berlin publishing, Germany.

    Google Scholar 

  • Furukawa S, Fujita T, Shimabukuro M, Iwaki M, Yamada Y, Nakajima Y, Nakayama O, Makishima M, Matsuda M, and Shimomura I (2004) Increased oxidative stress in obesity and its impact on metabolic syndrome. J Clin Invest 114, 1752–1761.

    CAS  Google Scholar 

  • Hsu CL and Yen GC (2008) Phenolic compounds: evidence for inhibitory effects against obesity and their underlying molecular signaling mechanisms. Mol Nutr Food Res 52, 53–61.

    Article  CAS  Google Scholar 

  • Oberley LW and Spitz DR (1984) Assay of SOD activity in tumor tissue. Methods Enzymol 105, 457–464.

    Article  CAS  Google Scholar 

  • Park J, Choe SS, Choi AH, Kim KH, Yoon MJ, Suganami T, Ogawa Y, and Kim JB (2006) Increase in glucose-6-phosphate dehydrogenase in adipocytes stimulates oxidative stress and inflammatory signals. Diabetes 55, 2939–2949.

    Article  CAS  Google Scholar 

  • Park J, Rho HK, Kim KH, Choe SS, Lee YS, and Kim JB (2005) Overexpression of glucose-6-phosphate dehydrogenase is associated with lipid dysregulation and insulin resistance in obesity. Mol Cell Biol 25, 5146–5157.

    Article  CAS  Google Scholar 

  • Rayalam S, Della-Fera MA, and Baile CA (2008) Phytochemicals and regulation of the adipocyte life cycle. J Nutr Biochem 19, 717–726.

    Article  CAS  Google Scholar 

  • Spiegelman BM and Flier JS (1999) Adipogenesis and obesity: rounding out the big picture. Cell 87, 377–389.

    Article  Google Scholar 

  • Student AK, Hsu RY, and Lane MD (1980) Induction of fatty acid synthetase synthesis in differentiating 3T3-L1 preadipocytes. J Biol Chem 225, 4745–4750.

    Google Scholar 

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Authors and Affiliations

  1. Department of Nutrition, University of Massachusetts, 01003, Amherst, MA, USA

    Ok-Hwan Lee & Young-Cheul Kim

  2. Department of Food and Nutrition, Hannam University, 306-791, Daejeon, Republic of Korea

    Young-In Kwon

  3. Traditional Food Research Group, Korea Food Research Institute, 463-746, Seongnam, Republic of Korea

    Hee-Do Hong

  4. Graduate School of Biotechnology Institute of Life Science and Resource, Kyung Hee University, 446-701, Suwon, Republic of Korea

    Cheon-Seok Park

  5. Graduate School of Complementary & Alternative Medicine, Pochon CHA University, 463-836, Seongnam, Republic of Korea

    Boo-Yong Lee

Authors
  1. Ok-Hwan Lee
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  2. Young-In Kwon
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  3. Hee-Do Hong
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  4. Cheon-Seok Park
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  5. Boo-Yong Lee
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  6. Young-Cheul Kim
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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

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