Inhibition of adhesion molecule expression on human venous endothelial cells by non-viral siRNA transfection
| Autor: | Hans Peter Wendel, Claudia Raabe, Liane Tetzloff, Gerhard Ziemer, Perikles Simon, Olaf Heidenreich, Albertus M. Scheule, Tobias Walker |
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| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
Small interfering RNA
Neuroscience Review Series Endothelium medicine.medical_treatment Intercellular Adhesion Molecule-1 venous graft disease coronary bypass grafting Biology Transfection Veins medicine Humans Saphenous Vein adhesion molecules RNA Small Interfering Cells Cultured Fluorescent Dyes Cell adhesion molecule Tumor Necrosis Factor-alpha Soluble cell adhesion molecules Endothelial Cells Cell Biology Adhesion Articles Molecular biology medicine.anatomical_structure Cytokine RNAi Molecular Medicine Endothelium Vascular Cell Adhesion Molecules Fluorescein-5-isothiocyanate |
| Zdroj: | Journal of Cellular and Molecular Medicine |
| ISSN: | 1582-4934 1582-1838 |
| Popis: | Objective: Expression of adhesion molecule receptors on venous endothelial cells crucially influences the fate of venous grafts by mediating leukocyte-endothelium interactions. These interactions include adhesion of leuko-cytes to the endothelium, followed by transendothelial migration, leading to neointimal hyperplasia (NIH) and finally graft occlusion. Therefore, inhibition of adhesion molecule expression may be a promising strategy to improve the quality of venous grafts.We tested the efficiency of non-viral transfection of human venous endothe-lial cells (HVEC) with short interfering RNA (siRNA) to specifically down-regulate adhesion molecule expression. Methods: Primary cultures of HVEC were examined for expression of the adhesion molecules ICAM1, VCAM1 and E-selectin (SELE) after non viral siRNA transfection. Adhesion molecule expression was measured by flow cytom-etry, real-time polymerase chain reaction and immunoblotting after stimulation with TNF-α, an inflammatory cytokine. Results: Non-transfected cells showed a strong increase of adhesion molecule expression following cytokine stimulation (P < 0.01). Upon transfection with specific siRNAs a sixfold decrease in ICAM1 (P < 0.001) and SELE expression and cell positivity (P < 0.05) and a twofold decrease in VCAM1 expression and cell positiv-ity (P < 0.01) Pcould be observed. SiRNA-mediated gene suppression of adhesion molecules was also reflected by corresponding decreases in adhesion protein and transcript levels. Conclusions: The expression of adhesion molecules on HVECs can be effectively inhibited by specific siRNAs using a safe, non-viral transfection approach. This is a promising tool to pre-condition venous bypass grafts in order to interfere with endothelium-leukocyte interactions and to prohibit neointima thickening or ath-erosclerosis, which are regarded to be the most important causes of venous graft failure. |
| Databáze: | OpenAIRE |
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