Efforts to overcome hypoxia condition in Balb/c mouse macrophages after intraperitoneal SRBC immunization

Medical Journal of Indonesia

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Title Efforts to overcome hypoxia condition in Balb/c mouse macrophages after intraperitoneal SRBC immunization
 
Creator Sarsanti, Pungguri Ayu Nega; Biomedical Sciences Graduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta
Sadikin, Mohamad; enter of Hypoxia and Oxidative Stress Study, Faculty of Medicine, Universitas Indonesia, Jakarta; Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
Jusman, Sri Widia Azraki; enter of Hypoxia and Oxidative Stress Study, Faculty of Medicine, Universitas Indonesia, Jakarta; Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
 
Subject cytoglobin; HIF-1α; HIF-2α; hypoxia; macrophage; PGC-1α
 
Description BACKGROUND Activated macrophages require increased oxygen to destroy foreign bodies, leading to an increase in the levels of reactive oxygen species (ROS). Therefore, macrophages would experience hypoxic and oxidative stress conditions at the same time. Thus, this study was aimed to evaluate the mechanism of the activated macrophages to overcoming this dual condition.METHODS The activated macrophages were harvested from the intraperitoneal cavities of 18 BALB/c mice immunized with 2% sheep red blood cells (SRBCs). The macrophage suspension was divided into four groups: control, 24, 48, and 72 hours after-immunization groups. The expressions of hypoxia-inducible factor (HIF)-1α, HIF-2α, and cytoglobin (Cygb), as markers for hypoxic condition, were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA), whereas peroxisome proliferator-activated receptor gamma coactivator (PGC-1α) protein as a marker for mitochondrial biogenesis and aerobic metabolism was measured with ELISA. The analysis of oxidative stress was conducted with the water-soluble tetrazolium salt test.RESULTS The HIF-1α mRNA expression was the highest at 24 hours, whereas the HIF-2α mRNA showed no increased expression during the observation. The Cygb mRNA decreased after 24 hours. The highest expressions of HIF-1α and HIF-2α proteins were detected at 72 hours, whereas the Cygb protein expression increased since 24 hours. The PGC-1α protein expression increased at 72 hours. The WST test showed the highest ROS level at 24 hours.CONCLUSIONS The macrophages were activated by SRBCs underwent dual hypoxia and oxidative stress condition simultaneously to overcome the foreign bodies. The macrophages overcame these stress conditions by increasing their aerobic metabolism.
 
Publisher Faculty of Medicine Universitas Indonesia
 
Contributor Direktorat Riset Dan Pengabdian Pada Masyarakat (DRPM) UI
 
Date 2019-05-08
 
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/1961
10.13181/mji.v28i1.1961
 
Source Medical Journal of Indonesia; Vol 28, No 1 (2019): March; 14-20
2252-8083
0853-1773
10.13181/mji.v28i1
 
Language eng
 
Relation http://mji.ui.ac.id/journal/index.php/mji/article/view/1961/1313
 
Rights Copyright (c) 2019 Pungguri Ayu Nega Sarsanti, Mohamad Sadikin, Sri Jusman
http://creativecommons.org/licenses/by-nc/4.0
 

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