Differential effects of ischemia/reperfusion on endothelial function and contractility in donation after circulatory death

https://doi.org/10.1016/j.healun.2019.03.004Get rights and content

BACKGROUND

Donation after circulatory death (DCD) could significantly improve cardiac graft availability. However, DCD hearts undergo potentially deleterious warm ischemia/reperfusion (I/R). As endothelial damage is a key factor in cardiac I/R injury, we aimed to investigate the tolerance of cardiac and endothelial function after various durations of warm ischemia to improve the timing and choice of cardioprotective therapies.

METHODS

Isolated, working rat hearts were perfused for 20 minutes aerobically, then underwent various periods of warm global ischemia and either 30 or 60 minutes of reperfusion.

RESULTS

Compared with non-ischemic hearts, recovery of left ventricular work (heart rate–developed pressure product) was significantly reduced at 60 minutes of reperfusion with ≥27 minutes of ischemia (p <0.05 for all), but was unchanged after 21 or 24 minutes of ischemia. Markers of cell death and edema significantly increased with ≥27-minute ischemia compared with non-ischemic hearts (p <0.05 for all). Endothelial-dependent vasodilation was significantly impaired compared with non-ischemic hearts with ≥24 minutes of ischemia, whereas endothelial-independent vasodilation was impaired with ≥27 minutes of ischemia (p <0.05 for all). Furthermore, with ≥24 minutes of ischemia, superoxide production by nitric oxide synthase and peroxynitrite levels were significantly increased compared with non-ischemic hearts, suggesting endothelial nitric oxide synthase (eNOS) uncoupling (p <0.05 for both).

CONCLUSIONS

The first signs of endothelial dysfunction after cardiac ischemia occur with less ischemia than cardiac functional alterations, and may result from increased eNOS uncoupling. Strategies aimed at improving eNOS coupling may thus help to optimize both endothelial and myocardial recovery, ultimately facilitating DCD heart transplantation.

Section snippets

Ethics

All experiments were carried out according to the European Convention for Animal Care and were approved by the Swiss animal welfare authorities and state veterinary office (Ethics Committee for Animal Experimentation [ECAE], Bern, Switzerland).

Isolated heart preparation

Adult male Wistar rats (384 ± 35 g; Janvier Labs, Le Genest-Saint-Isle, France), housed under standard conditions with unlimited access to food and water, were anesthetized intraperitoneally with 100 mg/kg ketamine (Narketan; Vetoquinol AG, Bern,

Baseline characteristics

Sample size and baseline characteristics for all heart perfusions are shown in Tables S1 and S2 (refer to Supplementary Material online). No difference among experimental groups was observed.

Post-ischemic functional recovery

Absolute values and percentage recovery of cardiac parameters are presented in Figure 2. Left ventricular work (LV work) was significantly lower for both absolute values (at 20, 40, and 60 minutes) and percentage recovery in hearts subjected to 27, 30, and 33 minutes of ischemia vs non-ischemic hearts (p

Discussion

In this study, we have demonstrated that endothelial dysfunction is more susceptible to warm global ischemia and reperfusion than cardiac dysfunction in an isolated rat heart model of DCD. The first signs of endothelial dysfunction were already present after 24 minutes of ischemia and reperfusion, whereas cardiac and smooth muscle cell (SMC) dysfunction appeared only after ischemic periods of 27 minutes. Endothelial dysfunction occurred in parallel with increases in NOS-dependent O2 and ONOO

Disclosure statement

The authors have no conflicts of interest to disclose. We thank Dr Daniel Rodriguez Gutierrez for the conceptual diagram illustration. This article/publication is based on work from COST Action EU-CARDIOPROTECTION CA16225, supported by the European Cooperation in Science and Technology. This study was supported by a project grant from the Swiss National Science Foundation (310030_149730/1).

References (68)

  • RR Giraldez et al.

    Decreased nitric-oxide synthase activity causes impaired endothelium-dependent relaxation in the postischemic heart

    J Biol Chem

    (1997)
  • T Münzel et al.

    Impact of oxidative stress on the heart and vasculature

    J Am Coll Cardiol

    (2017)
  • J Vásquez-Vivar

    Tetrahydrobiopterin, superoxide, and vascular dysfunction

    Free Rad Biol Med

    (2009)
  • R Fernández-Jiménez et al.

    Pathophysiology underlying the bimodal edema phenomenon after myocardial ischemia/reperfusion

    J Am Coll Cardiol

    (2015)
  • A Bernardo

    Effects of inflammatory cytokines on the release and cleavage of the endothelial cell-derived ultralarge von Willebrand factor multimers under flow

    Blood

    (2004)
  • MC Manukyan et al.

    Interleukin-10 protects the ischemic heart from reperfusion injury via the STAT3 pathway

    Surgery

    (2011)
  • J Gurevitch et al.

    Tumor necrosis factor-alpha is released from the isolated heart undergoing ischemia and reperfusion

    J Am Coll Cardiol

    (1996)
  • CW White et al.

    Physiologic changes in the heart following cessation of mechanical ventilation in a porcine model of donation after circulatory death: implications for cardiac transplantation: physiologic response to donor extubation

    Am J Transplant

    (2016)
  • P Niederberger et al.

    High pre-ischemic fatty acid levels decrease cardiac recovery in an isolated rat heart model of donation after circulatory death

    Metabolism

    (2017)
  • FJ Sutherland et al.

    The isolated blood and perfusion fluid perfused heart

    Pharmacol Res

    (2000)
  • SL Longnus et al.

    Heart transplantation with donation after circulatory determination of death

    Nat Rev Cardiol

    (2014)
  • S Osaki et al.

    The potential of cardiac allografts from donors after cardiac death at the University of Wisconsin Organ Procurement Organization

    Eur J Cardiothorac Surg

    (2010)
  • T Noterdaeme et al.

    What is the potential increase in the heart graft pool by cardiac donation after circulatory death?: potential of cardiac donation after circulatory death

    Transplant Int

    (2013)
  • CW White et al.

    Transplantation of hearts donated after circulatory death

    Front Cardiovasc Med

    (2018)
  • KK Dhital et al.

    Donation after circulatory death heart transplantation

    Curr Opin Organ Transplant

    (2017)
  • JH Lee et al.

    Coronary endothelial dysfunction and the index of microcirculatory resistance as a marker of subsequent development of cardiac allograft vasculopathy

    Circulation

    (2017)
  • MG Crespo-Leiro et al.

    Prevention and treatment of coronary artery vasculopathy

    Curr Opin Organ Transplant

    (2012)
  • G Heusch et al.

    The coronary circulation in cardioprotection: more than just one confounder

    Cardiovasc Res

    (2012)
  • AK Singhal et al.

    Role of endothelial cells in myocardial ischemia-reperfusion injury

    Vasc Dis Prevent

    (2010)
  • J Vinten-Johansen et al.

    Nitric oxide and the vascular endothelium in myocardial ischemia-reperfusion injury

    Ann NY Acad Sci

    (1999)
  • PS Tsao et al.

    Time course and mechanism of endothelial dysfunction in isolated ischemic- and hypoxic-perfused rat hearts

    Am J Physiol

    (1990)
  • H Otani

    The role of nitric oxide in myocardial repair and remodeling

    Antioxid Redox Signal

    (2009)
  • M Bartkevics et al.

    Efficacy of mechanical postconditioning following warm, global ischaemia depends on circulating fatty acid levels in an isolated, working rat heart model

    Eur J Cardiothorac Surg

    (2016)
  • E Farine et al.

    Controlled reperfusion strategies improve cardiac hemodynamic recovery after warm global ischemia in an isolated, working rat heart model of donation after circulatory death (DCD)

    Front Physiol

    (2016)
  • Cited by (16)

    • Mitochondrial integrity during early reperfusion in an isolated rat heart model of donation after circulatory death—consequences of ischemic duration

      2019, Journal of Heart and Lung Transplantation
      Citation Excerpt :

      For perfusion series 2, already at the time of tissue harvest (10 minutes reperfusion), a decrease of LV work was detectable with ≥ 27 minutes compared with no ischemia (p < 0.01 for both; Supplementary Figure S3B). Baseline characteristics for perfusion series 1 are reported elsewhere23 and for perfusion series 2 are presented in Supplementary Table 1. No differences between groups were observed.

    View all citing articles on Scopus
    View full text