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2010 Annual Meeting Abstracts

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Human Microvascular Endothelial Cells Treated with Hyperbaric Oxygen Become Resistant to Lethal Oxidation and Have Increased Expression of Cytoprotective Genes
George A. Perdrizet1, *Cassandra A. Godman2, *Charles Giardina2, *Lawrence E. Hightower2
1Morristown Memorial Hosptial, Morristown, NJ;2University of Connecticut, Storrs, CT

Objectives: 1. To test the hypothesis that exposure of human microvascular endothelial cells (HMEC-1 line) to hyperbaric oxygen treatment (HBOT), would increase resistance to lethal hyperthermia and oxidant stress compared to untreated cells. 2. To analyze changes in gene expression following HBOT exposure. Setting: Tissue culture laboratory. Patients: None; Design/Intervention: HMEC-1 cells exposed to one HBO2 treatment (100% oxygen at 2.4 atm for 1 hour), recovered and then challenged with lethal hyperthermia (47ºC) or a strong oxidizing agent (t-butyl hydroperoxide). Viability determined by propidium iodine staining or MTS assay. Cell cultures were sampled for RNA expression using an Illumina® microarray representing 38,275 unique sequences. Results: HBOT significantly increased resistance of HMEC-1 cells to lethal heat stress compared to untreated cells. Cell death occurred in 59.8% of untreated cells versus 5.2% in the HBOT cells, p <0.05. Likewise, cell survival was greater for HBOT cells compared to control cells at all concentrations of t-butyl hydroperoxide tested (Figure). HMEC-1 cells exposed to HBOT demonstrated a significant change in expression in 21.4% (8,181 genes) of the total number of genes tested at 24 hours after HBOT. Most notably, three major pathways were up-regulated; Nrf2-mediated oxidative stress response, Integrin signaling, and ERK/MAPK signaling. Conclusion: A single HBOT exposure of HMEC-1 cells provides protection against lethal hyperthermia and severe oxidant stress. Changes in gene expression are consistent with the observed phenotypic changes and suggest that HBOT maybe a clinically relevant agent to provide pre-operative stress conditioning.

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