Transfection of short-interfering RNA silences adhesion molecule expression on cardiac microvascular cells
1Clinic for Thoracic, Cardiac and Vascular Surgery, University of Tübingen, Tübingen, Germany. tobias.walker@med.uni-tuebingen.de
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.
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