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Thorac Cardiovasc Surg 2011; 59(6): 322-328
DOI: 10.1055/s-0030-1271142
Original Basic Science

© Georg Thieme Verlag KG Stuttgart · New York

Transfection of Short-Interfering RNA Silences Adhesion Molecule Expression on Cardiac Microvascular Cells

T. Walker1 , E. Saup1 , A. Nolte2 , P. Simon3 , A. Kornberger1 , V. Steger1 , O. Adolph4 , H.-P. Wendel2
  • 1Clinic for Thoracic, Cardiac and Vascular Surgery, University of Tübingen, Tübingen, Germany
  • 2Div. of Congenital & Paediatric Cardiac Surgery, Clinical Research Laboratory, Children's Hospital, University of Tübingen, Tübingen, Germany
  • 3Department of Sports Medicine, Rehabilitation and Disease Prevention, University of Mainz, Mainz, Germany
  • 4Department of Anesthesiology, University Ulm, Ulm, Germany
Further Information

Publication History

received January 1, 2011 resubmitted April 1, 2011

accepted April 2, 2011

Publication Date:
20 June 2011 (online)

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Abstract

Background: Acute rejection reactions and the development of graft arterial disease are serious limitations after transplantation. Both are connected to the expression of adhesion molecules on the activated microvascular endothelium of the allograft. Methods: siRNA-mediated silencing of ESELE, ICAM-1, and VCAM-1 on human cardiac microvascular cells (HCMEC) was investigated in order to inhibit leukocyte-endothelial interactions. HCMEC were investigated for the time-dependent expression of ESELE, ICAM-1, and VCAM-1 after TNF-α stimulation and for siRNA mediated suppression using a nonviral transfecting approach. Furthermore, the effects of siRNA transfection on leukocyte binding to the endothelium were analyzed. Results: Transfection with siRNA induced a significant suppression of adhesion molecule expression, regardless of whether there had been a prior single or cocktail transfection of the sequences (p < 0.05). The quantity of attaching leukocytes was significantly reduced after an equal silencing of adhesion molecules (p < 0.05). Conclusions: This investigation demonstrates that liposomal transfection of HCMEC with specific siRNA sequences is capable of both repressing adhesion molecule expression and of reducing subsequent leukocyte-endothelial actions.

Key words

transplantation - heart - endothelium (includes related subject matter) - ischemia/reperfusion

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Dr. Tobias Walker

Clinic for Thoracic, Cardiac and Vascular Surgery
University of Tübingen

Hoppe-Seyler-Straße 3

72076 Tübingen

Germany

Phone: +49 7 07 12 98 32 88

Fax: +49 70 71 29 53 69

Email: tobias.walker@med.uni-tuebingen.de