NESS - 2010 Annual Meeting Abstracts: Human Microvascular Endothelial Cells Treated with Hyperbaric Oxygen Become Resistant to Lethal Oxidation and Have Increased Expression of Cytoprotective Genes

2010 Annual Meeting Abstracts

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 HBO₂treatment (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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