Oxidative stress provokes atherogenic changes in adipokine gene expression in 3T3-L1 adipocytes

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Abstract

Increased oxidative stress has been associated with obesity-related disorders. In this study, we investigated how oxidative stress, in different ways of exposure, regulates gene expression of various adipokines in 3T3-L1 adipocytes. Exposure to 100–500 μM H2O2 for 10 min, as well as exposure to 5–25 mU/ml glucose oxidase for 18 h, similarly decreased adiponectin, leptin, and resistin mRNAs, and increased plasminogen activator inhibitor-1 mRNA. Secretion levels of adipokines were also changed by oxidative stress in parallel with mRNA expression levels. Although a peak increase in plasminogen activator inhibitor-1 mRNA was achieved between 4 and 8 h after exposure to H2O2 for 10 min, significant decreases in adiponectin and resistin mRNA were observed after 16 h, while leptin mRNA was decreased earlier. Our results suggest that oxidative stress, even of short duration, has a significant impact on the regulation of various adipokine gene expressions favoring atherosclerosis.

Section snippets

Materials and methods

Materials. Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, d-biotin, insulin, 3-isobutyl-1-methylxanthine, dexamethasone, bovine serum albumin (BSA), and all other cell culture reagents were obtained from Sigma (Saint Louis, MO) unless otherwise mentioned. Fetal bovine serum (FBS) was purchased from Equitech-bio (Kerrville, TX) and the same lot of FBS was used throughout the study. Glucose oxidase (GO), H2O2, N-acetyl-l-cysteine (NAC), and all other chemicals were also

Sustained exposure to H2O2 decreases adiponectin, leptin, and resistin mRNA levels, and increases the PAI-1 mRNA level

To examine the effects of H2O2 on adipokine gene expressions, 3T3-L1 adipocytes were incubated with various concentrations of GO in the presence of glucose for 18 h under serum-free conditions. In the presence of 25 mU/ml GO, the H2O2 concentration was reportedly 14.8 ± 1.5 μM at 30 min after GO addition [25]. Total RNA was extracted from the cells and mRNA levels of adiponectin, leptin, resistin, and PAI-1 were measured using quantitative RT-PCR. Adiponectin, leptin, and resistin mRNA levels were

Discussion

In the present study, we demonstrated that oxidative stress decreased adiponectin, leptin, and resistin mRNA expressions and increased PAI-1 mRNA expression in 3T3-L1 adipocytes. The secretion level of each adipokine was changed in parallel with mRNA level by the oxidative stress.

By modifying the method parameters of previous studies [23], [24], we showed that both exposure to high-level H2O2 for a short time and exposure to low-level H2O2 for a long time similarly changed mRNA expression of

Acknowledgment

We are grateful to Miss Kana Yoshikuni for her technical assistance.

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