Methylprednisolone and Tacrolimus Prevent Hypothermia-Induced Endothelial Dysfunction

Diestel, Antje; Billecke, Nils; Roessler, Joerg; Schmitt, Boris; Troeller, Silke; Schwartlander, Ruth; Berger, Felix; Sauer, Igor Maximilian; Schmitt, Katharina Rose Luise

doi:10.1016/j.healun.2009.04.003

« Previous Next »The Journal of Heart and Lung Transplantation
Volume 28, Issue 7 , Pages 718-724, July 2009

Methylprednisolone and Tacrolimus Prevent Hypothermia-Induced Endothelial Dysfunction

Antje Diestel, PhD
- Department of Pediatric Cardiology, Charité-Universitätsmedizin Berlin, Berlin, Germany
Nils Billecke, MSc
- Department of General, Visceral and Transplantation Surgery, Charité-Universitätsmedizin Berlin, Berlin, Germany
Joerg Roessler, MS
- Department of Congenital Heart Disease/Pediatric Cardiology, Deutsches Herzzentrum Berlin, Berlin, Germany
Boris Schmitt, MD
- Department of Pediatric Cardiology, Charité-Universitätsmedizin Berlin, Berlin, Germany
Silke Troeller, MSc
- Department of Congenital Heart Disease/Pediatric Cardiology, Deutsches Herzzentrum Berlin, Berlin, Germany
Ruth Schwartlander, PhD
- Department of General, Visceral and Transplantation Surgery, Charité-Universitätsmedizin Berlin, Berlin, Germany
Felix Berger, MD, PhD
- Department of Pediatric Cardiology, Charité-Universitätsmedizin Berlin, Berlin, Germany
- Department of Congenital Heart Disease/Pediatric Cardiology, Deutsches Herzzentrum Berlin, Berlin, Germany
Igor Maximilian Sauer, MD, PhD
- Department of General, Visceral and Transplantation Surgery, Charité-Universitätsmedizin Berlin, Berlin, Germany
- These authors contributed equally to this work.
Katharina Rose Luise Schmitt, MD
- Department of Congenital Heart Disease/Pediatric Cardiology, Deutsches Herzzentrum Berlin, Berlin, Germany
- These authors contributed equally to this work.
- Reprint requests: Katharina Rose Luise Schmitt, MD, Department of Congenital Heart Disease/Pediatric Cardiology, Deutsches Herzzentrum Berlin, Augustenburger Platz 1, 13353 Berlin, Germany. Telephone: 49-30-45-93-2800. Fax: 49-30-45-93-2900

Received 5 December 2008; received in revised form 10 March 2009; accepted 7 April 2009. published online 15 May 2009.

Background

Hypothermia is used to preserve organs for transplantation and is the oldest method to protect organs during complex pediatric cardiac surgery. Loss of tissue function and tissue edema are common complications in children undergoing corrective cardiac surgery and heart transplantation. The present study was designed to examine the effects of methylprednisolone and tacrolimus on endothelial cell function and morphology after deep hypothermia and rewarming.

Methods

Human umbilical vein endothelial cells were pre-treated with methylprednisolone or tacrolimus, or both, incubated within a specially designed bioreactor or in monolayers, and then exposed to a dynamic cooling and rewarming protocol. Immunocytochemistry, time-lapse video microscopy, cell permeability and adherence assays, and Western blot analysis were performed.

Results

Confluent endothelial cells exposed to hypothermia displayed elongated cell shapes with intercellular gap formation, increased endothelial cell-layer permeability, and loss in adherence. Upon rewarming, however, endothelial cell integrity was restored. Opening and closing of intercellular gaps was dependent on extracellular signal-regulated kinase 1 and 2 (ERK 1/2) activation and connexin 43 expression. The combined treatment with methylprednisolone and tacrolimus inhibited these hypothermia-induced changes.

Conclusions

These results suggest that methylprednisolone and tacrolimus inhibit hypothermia-induced endothelial gap formation by phosphorylated ERK 1/2 inhibition and connexin 43 stabilization. Application of combined drugs that affect multiple targets may therefore be considered as a possible new therapeutic strategy to prevent endothelial dysfunction after hypothermia and rewarming.