Relative hypoxia and oxidative stress in spleen lymphocytes of immunized Balb/c mice as indicated by HIF-1α, HIF-2α, Nrf2 expression, and glutathione peroxidase activity

Medical Journal of Indonesia

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Title Relative hypoxia and oxidative stress in spleen lymphocytes of immunized Balb/c mice as indicated by HIF-1α, HIF-2α, Nrf2 expression, and glutathione peroxidase activity
 
Creator Praditi, Citra; Master Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jakarta
Prijanti, Ani R.; Center of Hypoxia and Oxidative Stress Studies, Department of Biochemistry and Molecular Biology, Faculty of Medicine, Jakarta
Jusman, Sri W.A.; Center of Hypoxia and Oxidative Stress Studies, Department of Biochemistry and Molecular Biology, Faculty of Medicine, Jakarta
Sadikin, Mohamad; Center of Hypoxia and Oxidative Stress Studies, Department of Biochemistry and Molecular Biology, Faculty of Medicine, Jakarta
 
Subject HIF-1α; HIF-2α; Nrf2; oxidative stress; relative hypoxia
 
Description Background: Lymphocytes activated by immunization must increase their metabolism to meet the energy requirements for mitosis, differentiation, and protein synthesis, which may subject the cell to conditions of relative hypoxia and oxidative stress. This study was conducted to investigate the increase in the levels of transcription factors involved in both conditions.Methods: Male Balb/c mice were divided into the following four groups, each consisting of six animals: the control and three experimental groups. The experimental groups were immunized by injection of 0.2 ml of 2% sheep red blood cells (SRBC) suspended in phosphate-buffered saline (PBS). Lymphocytes were harvested from the spleens of each group at time intervals of 24-, 48-, and 72-h post-immunization. The buffy coat from splenocytes was separated using Ficoll Histopaque as the medium. The lymphocytes were separated from adherent cells by incubating the purified splenocytes in microtubes for 2-h. Cells were lysed by three freeze–thaw cycles (−80°C and 37°C) and used to analyze the levels of HIF-1α and HIF-2α (mRNA and protein), Nrf2 (protein), and glutathione peroxidase (GPx) activity.Results: The treatment caused an increase in GPx activity and HIF-1α protein concentration 24-h post-immunization, whereas the HIF-1α mRNA levels remained static. Elevated Nrf2 protein levels were detected within 48-h after treatment. Meanwhile, the HIF-2α mRNA and protein levels increased within72-h after immunization.Conclusion: Immunization with SRBC suspension induced relative hypoxia, elevated reactive oxygen species (ROS), and oxidative stress in the lymphocytes as indicated by the increase in both HIF-1α and HIF-2α protein and mRNA levels, GPx activity, and Nrf2 protein levels.
 
Publisher Faculty of Medicine Universitas Indonesia
 
Contributor
 
Date 2018-12-31
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion

 
Format application/pdf
 
Identifier http://mji.ui.ac.id/journal/index.php/mji/article/view/2152
10.13181/mji.v27i4.2152
 
Source Medical Journal of Indonesia; Vol 27, No 4 (2018): December; 223-8
2252-8083
0853-1773
10.13181/mji.v27i4
 
Language eng
 
Relation http://mji.ui.ac.id/journal/index.php/mji/article/view/2152/1256
 
Rights Copyright (c) 2018 Citra Praditi, Ani R. Prijanti, Sri W.A. Jusman, Mohamad Sadikin
http://creativecommons.org/licenses/by-nc/4.0
 

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