Elevated extracellular CO2 level affects the adaptive transcriptional response and survival of human peripheral blood mononuclear cells toward hypoxia and oxidative stress

  • Septelia Inawati Wanandi Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; Molecular Biology and Proteomics Core Facilities, Indonesian Medical and Education Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-7963-8853
  • Sekar Arumsari Molecular Biology and Proteomics Core Facilities, Indonesian Medical and Education Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-3623-9617
  • Edwin Afitriansyah Molecular Biology and Proteomics Core Facilities, Indonesian Medical and Education Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-7388-7269
  • Resda Akhra Syahrani Molecular Biology and Proteomics Core Facilities, Indonesian Medical and Education Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Idham Rafly Dewantara Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Luthfian Aby Nurachman Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Ihya Fakhrurizal Amin Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Putera Dewa Haryono Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Kenny Budiman Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0003-0534-784X
  • Adrianus Jonathan Sugiharta Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-6611-3070
  • Amino Aytiwan Remedika Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Farhan Hilmi Tafikulhakim Undergraduate Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Febriana Catur Iswanti Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; Molecular Biology and Proteomics Core Facilities, Indonesian Medical and Education Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Jason Youngbin Lee Biomedical Science Master Program, Rutgers-Robert Wood Johnson Medical School, New Jersey, USA; USAID Research Innovation Fellowship 2017, Washington DC, USA
  • Debabrata Banerjee Department of Pharmacology, Rutgers-Robert Wood Johnson Medical School, New Jersey, USA
Keywords: elevated CO2, extracelullar pH, oxidative stress, PBMC, reactive oxygen species
Abstract viewed: 556 times
PDF downloaded: 309 times
HTML downloaded: 53 times
EPUB downloaded: 89 times

Abstract

BACKGROUND High carbon dioxide (CO2) level from indoor environments, such as classrooms and offices, might cause sick building syndrome. Excessive indoor CO2 level increases CO2 level in the blood, and over-accumulation of CO2 induces an adaptive response that requires modulation of gene expression. This study aimed to investigate the adaptive transcriptional response toward hypoxia and oxidative stress in human peripheral blood mononuclear cells (PBMCs) exposed to elevated CO2 level in vitro and its association with cell viability.

METHODS PBMCs were treated in 5% CO2 and 15% CO2, representatives a high CO₂ level condition for 24 and 48 hours. Extracellular pH (pHe) was measured with a pH meter. The levels of reactive oxygen species were determined by measuring superoxide and hydrogen peroxide with dihydroethidium and dichlorofluorescin-diacetate assay. The mRNA expression levels of hypoxia-inducible factor (HIF)-1α, HIF-2α, nuclear factor (NF)-κB, and manganese superoxide dismutase (MnSOD) were analyzed using a real-time reverse transcriptase-polymerase chain reaction (qRT-PCR). Cell survival was determined by measuring cell viability.

RESULTS pHe increased in 24 hours after 15% CO₂ treatment, and then decreased in 48 hours. Superoxide and hydrogen peroxide levels increased after the 24- and 48-hour of high CO₂ level condition. The expression levels of NF-κB, MnSOD, HIF-1α, and HIF-2α decreased in 24 hours and increased in 48 hours. The increased antioxidant mRNA expression in 48 hours showed that the PBMCs were responsive under high CO2 conditions. Elevated CO2 suppressed cell viability significantly in 48 hours.

CONCLUSIONS After 48 hours of high CO₂ level condition, PBMCs showed an upregulation in genes related to hypoxia and oxidative stress to overcome the effects of CO2 elevation.

References

  1. Bierwirth PN. Carbon dioxide toxicity and climate change: a major unapprehended risk for human health. Web Published: Research Gate; 2018. https://doi.org/10.13140/RG.2.2.16787.48168

  2. Franswijaya CC, Kusnoputranto H. Indoor air quality and sick building syndrome in the 4th building of BPS Headquarters, Central Jakarta, in 2012. Universitas Indonesia: Faculty of Public Health; 2018. Indonesian.

  3. Patel S, Ghadimi M, Majmundar SH. Physiology, carbon dioxide retention [Internet]. Treasure Island (FL): StatPearls Publishing; 2020 [updated 2020 Jan 13; cited 2020 Feb 18]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK482456/4.

  4. Casalino-Matsuda SM, Wang N, Ruhoff PT, Matsuda H, Nlend MC, Nair A, et al. Hypercapnia alters expression of immune response, nucleosome assembly and lipid metabolism genes in differentiated human bronchial epithelial cells. Sci Rep. 2018;8(1):13508. https://doi.org/10.1038/s41598-018-32008-x

  5. Span PN, Bussink J. Biology of hypoxia. Semin Nucl Med. 2015;45(2):101-9. https://doi.org/10.1053/j.semnuclmed.2014.10.002

  6. Masoud GN, Li W. HIF-1α pathway: role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 2015;5(5):378-89. https://doi.org/10.1016/j.apsb.2015.05.007

  7. Sørensen BS, Busk M, Overgaard J, Horsman MR, Alsner J. Simultaneous hypoxia and low extracellular pH suppress overall metabolic rate and protein synthesis in vitro. PLoS One. 2015;10(8):e0134955. https://doi.org/10.1371/journal.pone.0134955

  8. Kelly FJ, Fussell JC. Air pollution and airway disease. Clin Exp Allergy. 2011;41(8):1059-71. https://doi.org/10.1111/j.1365-2222.2011.03776.x

  9. Morgan MJ, Liu ZG. Crosstalk of reactive oxygen species and NF-κB signaling. Cell Res. 2011;21(1):103-15. https://doi.org/10.1038/cr.2010.178

  10. Abbas AK, Lichtman AH, Pillai S. Cellular and molecular immunology. 8th ed. Philadelphia: Elsevier; 2015. p. 14-25.

  11. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25(4):402-8. https://doi.org/10.1006/meth.2001.1262

  12. Swietach P, Vaughas-Jones RD, Harris AL. Regulation of tumor pH and the role of carbonic anhydrase 9. Cancer Metastasis Rev. 2007;26(2):299-310. https://doi.org/10.1007/s10555-007-9064-0

  13. Damaghi M, Wojtkowiak, JW, Gillies RJ. pH sensing and regulation in cancer. Front Physiol. 2013;4:370. https://doi.org/10.3389/fphys.2013.00370

  14. Filatova A, Seidel S, Böğürcü N, Gräf S, Garvalov BK, Acker T. Acidosis acts through HSP90 in a PHD/VHL-independent manner to promote HIF function and stem cell maintenance in glioma. Cancer Res. 2016;76(19):5845-56. https://doi.org/10.1158/0008-5472.CAN-15-2630

  15. Komlódi T, Geibl FF, Sassani M, Ambrus A, Tretter L. Membrane potential and delta pH dependency of reverse electron transport-associated hydrogen peroxide production in brain and heart mitochondria. J Bioenerg Biomembr. 2018;50:355-65. https://doi.org/10.1007/s10863-018-9766-8

  16. Kelley EE, Khoo NK, Hundley NJ, Malik UZ, Freeman BA, Tarpey MM. Hydrogen peroxide is the major oxidant product of xanthine oxidase. Free Radic Biol Med. 2010;48(4):493-8. https://doi.org/10.1016/j.freeradbiomed.2009.11.012

  17. Riemann A, Schneider B, Ihling A, Nowak M, Sauvant C, Thews O, et al. Acidic environment leads to ROS-induced MAPK signaling in cancer cells. PloS One. 2011;6(7):e22445. https://doi.org/10.1371/journal.pone.0022445

  18. Hoesel B, Schmid JA. The complexity of NF-kB signaling in inflammation and cancer. Mol Cancer. 2013;12:86. https://doi.org/10.1186/1476-4598-12-86

  19. Luks AM. Physiology in medicine: a physiologic approach to prevention and treatment of acute high-altitude illnesses. J Appl Physiol. 2015;118(5):509-19. https://doi.org/10.1152/japplphysiol.00955.2014

  20. Taylor CT, Cummins EP. Regulation of gene expression by carbon dioxide. J Physiol. 2011;598(Pt 4):797-803. https://doi.org/10.1113/jphysiol.2010.201467

  21. Selfridge AC, Cavadas MA, Scholz CC, Campbell EL, Welch LC, Lecuona E, et al. Hypercapnia suppresses the HIF-dependent adaptive response to hypoxia. J Biol Chem. 2016;291(22):11800-8. https://doi.org/10.1074/jbc.M116.713941

Published
2021-03-25
How to Cite
1.
Wanandi SI, Arumsari S, Afitriansyah E, Syahrani RA, Dewantara IR, Nurachman LA, Amin IF, Haryono PD, Budiman K, Sugiharta AJ, Remedika AA, Tafikulhakim FH, Iswanti FC, Lee JY, Banerjee D. Elevated extracellular CO<sub>2</sub&gt; level affects the adaptive transcriptional response and survival of human peripheral blood mononuclear cells toward hypoxia and oxidative stress. Med J Indones [Internet]. 2021Mar.25 [cited 2021Oct.26];30(1):5-12. Available from: https://mji.ui.ac.id/journal/index.php/mji/article/view/3810
Section
Basic Medical Research