Different effects of antisense RelA p65 and NF-κB1 p50 oligonucleotides on the nuclear factor-κB mediated expression of ICAM-1 in human coronary endothelial and smooth muscle cells

Smooth muscle cells (SMC) from human coronary plaque material (HCPSMC, plaque material of 52 patients), SMC from the human coronary media (HCMSMC), human endothelial cells (EC) from umbilical veins (HUVEC), and human coronary EC (HCAEC) were successfully isolated (HCPSMC, HUVEC), identified and cultured (HCPSMC, HCMSMC, HUVEC, HCAEC). 12 hrs prior to TNF-α stimulus (20 ng/mL, 6 hrs) RelA p65 and NF-κB1 p50 (1, 2, 4, 10, 20, and 30 μM) and controls were added for a period of 18 hrs. In HUVEC and HCAEC there was a dose dependent inhibition of ICAM-1 expression after adding of both RelA p65 and NF-κB1 p50. No inhibitory effect was seen after incubation of HCMSMC with RelA p65 and NF-κB1 p50. A moderate inhibition of ICAM-1 expression was found after simultaneous addition of RelA p65 and NF-κB1 p50 to HCPSMC, no inhibitory effect was detected after individual addition of RelA p65 and NF-κB1 p50.The data point out that differences exist in the NF-κB mediated expression of ICAM-1 between EC and SMC. Experimental antisense strategies directed against RelA p65 and NF-κB1 p50 in early atherosclerosis and restenosis are promising in HCAEC but will be confronted with redundant pathways in HCMSMC and HCPSMC.Atherosclerosis is currently considered to be an exaggerated response of the vessel wall to injury characterized by inflammation and fibrocellular proliferation [1]. This view is supported by the demonstration of abundant macrophages and T lymphocytes in atherosclerotic plaques that accumulate because of adhesion molecule expression [2-4]. Nuclear factor-κB (NF-κB) regulates a variety of genes coding for cytokines [5-9] and adhesion receptors [8], that mediate endothelium-leukocyte adhesion [10]. NF-κB-regulated gene products such as interleukin-lβ (IL-1β), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1) have been found in tissue specimens of atherosclerotic lesions [1]. Activated NF-κB was identifie