Abstract
BackgroundAs organ shortage is increasing, the acceptance of marginal donors increases, which might result in poor organ function and patient survival. Mostly, organ damage is caused during brain death (BD), cold ischemic time (CIT) or after reperfusion due to oxidative stress or the induction of apoptosis. The aim of this study was to study a panel of genes involved in oxidative stress and apoptosis and compare these findings with immunohistochemistry from a BD and living donation (LD) pig model and after cold ischemia time (CIT).MethodsBD was induced in pigs; after 12 h organ retrieval was performed; heart, liver and kidney tissue specimens were collected in the BD (n = 6) and in a LD model (n = 6). PCR analysis for NFKB1, GSS, SOD2, PPAR-alpha, OXSR1, BAX, BCL2L1, and HSP 70.2 was performed and immunohistochemistry used to show apoptosis and nitrosative stress induced cell damage.ResultsIn heart tissue of BD BAX, BCL2L1 and HSP 70.2 increased significantly after CIT. Only SOD2 was over-expressed after CIT in BD liver tissue. In kidney tissue, BCL2L1, NFKB, OXSR1, SOD2 and HSP 70.2 expression was significantly elevated in LD. Immunohistochemistry showed a significant increase in activated Caspase 3 and nitrotyrosine positive cells after CIT in BD in liver and in kidney tissue but not in heart tissue.ConclusionThe up-regulation of protective and apoptotic genes seems to be divergent in the different organs in the BD and LD setting; however, immunohistochemistry revealed more apoptotic and nitrotyrosine positive cells in the BD setting in liver and kidney tissue whereas in heart tissue both BD and LD showed an increase.
Highlights
As organ shortage is increasing, the acceptance of marginal donors increases, which might result in poor organ function and patient survival
In brain death (BD) animals BCl2 associated X protein (BAX), B-cell lymphoma 2 like 1 (BCL2L1) and 70.2 Heat shock protein 70.2 (HSP) 70.2 expression increased after cold ischemia time (CIT) compared to pre perfusion (2.7 fold, p < 0.05, 3 fold, p < 0.05 and 5.9 fold, p < 0.001 respectively)
The number of activated Caspase 3 positive cells and nitrotyrosine positive cells increased significantly after perfusion and after 4 h of CIT in heart tissue of both BD and living donation (LD) donor organs compared to the samples taken prior to perfusion
Summary
As organ shortage is increasing, the acceptance of marginal donors increases, which might result in poor organ function and patient survival. The aim of this study was to study a panel of genes involved in oxidative stress and apoptosis and compare these findings with immunohistochemistry from a BD and living donation (LD) pig model and after cold ischemia time (CIT). The aim of this study was to study a panel of genes involved in oxidative stress and apoptosis and correlate gene expression with immunohistochemical findings of apoptosis and nitrosative stress and serum markers of oxidative stress before and after harvesting of organs from a BD and LD pig model and after cold ischemia time (CIT) in order to have the potential possibility to find novel strategies for donor pre-conditioning and amelioration in further experiments
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