Abstract
A genome-wide microarray analysis of gene expression was carried out on human microvascular endothelial cells (HMEC-1) exposed to hyperbaric oxygen treatment (HBOT) under conditions that approximated clinical settings. Highly up-regulated genes included immediate early transcription factors (FOS, FOSB, and JUNB) and metallothioneins. Six molecular chaperones were also up-regulated immediately following HBOT, and all of these have been implicated in protein damage control. Pathway analysis programs identified the Nrf-2-mediated oxidative stress response as one of the primary responders to HBOT. Several of the microarray changes in the Nrf2 pathway and a molecular chaperone were validated using quantitative PCR. For all of the genes tested (Nrf2, HMOX1, HSPA1A, M1A, ACTC1, and FOS), HBOT elicited large responses, whereas changes were minimal following treatment with 100% O2 in the absence of elevated pressure. The increased expression of immediate early and cytoprotective genes corresponded with an HBOT-induced increase in cell proliferation and oxidative stress resistance. In addition, HBOT treatment enhanced endothelial tube formation on Matrigel plates, with particularly dramatic effects observed following two daily HBO treatments. Understanding how HBOT influences gene expression changes in endothelial cells may be beneficial for improving current HBOT-based wound-healing protocols. These data also point to other potential HBOT applications where stimulating protection and repair of the endothelium would be beneficial, such as patient preconditioning prior to major surgery.
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Acknowledgements
We would like to thank OxyHeal Health Group for providing the funding and hyperbaric chamber for these studies. We would also like to extend our gratitude to Alexandria Thornton and Bryan Lawlor for their work during the early stages of the project. We would like to thank Advanced Turbine Services, LLC for donating some of the equipment used in this work. For technical support, we thank Dr. Carol Norris, Facility Scientist at the University of Connecticut FCCM Facility.
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Supplementary Table 1
Gene list of biological functions for HBOT0 (XLS 50 kb)
Supplementary Table 2
Gene list of biological functions for HBOT24 (XLS 99 kb)
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Godman, C.A., Chheda, K.P., Hightower, L.E. et al. Hyperbaric oxygen induces a cytoprotective and angiogenic response in human microvascular endothelial cells. Cell Stress and Chaperones 15, 431–442 (2010). https://doi.org/10.1007/s12192-009-0159-0
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DOI: https://doi.org/10.1007/s12192-009-0159-0