Endothelial dysfunction of bypass graft: Direct comparison of In Vitro and In Vivo models of ischemia-reperfusion injury

Gábor Veres, Péter Hegedus, Eniko Barnucz, Raphael Zöller, Stephanie Klein, Harald Schmidt, T. Radovits, Sevil Korkmaz, Matthias Karck, G. Szabó

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Although, ischemia/reperfusion induced vascular dysfunction has been widely described, no comparative study of in vivo- and in vitro-models exist. In this study, we provide a direct comparison between models (A) ischemic storage and in-vitro reoxygenation (B) ischemic storage and in vitro reperfusion (C) ischemic storage and in-vivo reperfusion. Methods and Results: Aortic arches from rats were stored for 2 hours in saline. Arches were then (A) in vitro reoxygenated (B) in vitro incubated in hypochlorite for 30 minutes (C) in vivo reperfused after heterotransplantation (2, 24 hours and 7 days reperfusion). Endothelium-dependent and independent vasorelaxations were assessed in organ bath. DNA strand breaks were assessed by TUNEL-method, mRNA expressions (caspase-3, bax, bcl-2, eNOS) by quantitative real-time PCR, proteins by Western blot analysis and the expression of CD-31 by immunochemistry. Endothelium-dependent maximal relaxation was drastically reduced in the in-vivo models compared to ischemic storage and in-vitro reperfusion group, and no difference showed between ischemic storage and control group. CD31-staining showed significantly lower endothelium surface ratio in-vivo, which correlated with TUNEL-positive ratio. Increased mRNA and protein levels of pro- and anti-apoptotic gens indicated a significantly higher damage in the in-vivo models. Conclusion: Even short-period of ischemia induces severe endothelial damage (in-vivo reperfusion model). In-vitro models of ischemia-reperfusion injury can be limitedly suited for reliable investigations. Time course of endothelial stunning is also described.

Original languageEnglish
Article numbere0124025
JournalPLoS One
Volume10
Issue number4
DOIs
Publication statusPublished - Apr 15 2015

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ischemia
Reperfusion Injury
Grafts
Reperfusion
Transplants
endothelium
Endothelium
Arches
In Situ Nick-End Labeling
immunochemistry
hypochlorites
Ischemia
Hypochlorous Acid
Messenger RNA
vasodilation
Immunochemistry
caspase-3
Apoptosis Regulatory Proteins
DNA Breaks
in vivo studies

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Endothelial dysfunction of bypass graft : Direct comparison of In Vitro and In Vivo models of ischemia-reperfusion injury. / Veres, Gábor; Hegedus, Péter; Barnucz, Eniko; Zöller, Raphael; Klein, Stephanie; Schmidt, Harald; Radovits, T.; Korkmaz, Sevil; Karck, Matthias; Szabó, G.

In: PLoS One, Vol. 10, No. 4, e0124025, 15.04.2015.

Research output: Contribution to journalArticle

Veres, Gábor ; Hegedus, Péter ; Barnucz, Eniko ; Zöller, Raphael ; Klein, Stephanie ; Schmidt, Harald ; Radovits, T. ; Korkmaz, Sevil ; Karck, Matthias ; Szabó, G. / Endothelial dysfunction of bypass graft : Direct comparison of In Vitro and In Vivo models of ischemia-reperfusion injury. In: PLoS One. 2015 ; Vol. 10, No. 4.
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