Voisard R, Huber N, Baur R, Susa M, Ickrath O, Both A, Koenig W, Hombach V
Department of Internal Medicine II - Cardiology, University of Ulm, Robert-Kochstrasse 8, D-89081 Ulm, Germany.
BMC Mol Biol. 2001;2:7. doi: 10.1186/1471-2199-2-7. Epub 2001 Aug 8.
Activation of nuclear factor-kappaB (NF-kappaB) is one of the key events in early atherosclerosis and restenosis. We hypothesized that tumor necrosis factor-alpha (TNF-alpha) induced and NF-kappaB mediated expression of intercellular adhesion molecule-1 (ICAM-1) can be inhibited by antisense RelA p65 and NF-kappaB1 p50 oligonucleotides (RelA p65 and NF-kappaB1 p50).
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-alpha stimulus (20 ng/mL, 6 hrs) RelA p65 and NF-kappaB1 p50 (1, 2, 4, 10, 20, and 30 microM) 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-kappaB1 p50. No inhibitory effect was seen after incubation of HCMSMC with RelA p65 and NF-kappaB1 p50. A moderate inhibition of ICAM-1 expression was found after simultaneous addition of RelA p65 and NF-kappaB1 p50 to HCPSMC, no inhibitory effect was detected after individual addition of RelA p65 and NF-kappaB1 p50.
The data point out that differences exist in the NF-kappaB mediated expression of ICAM-1 between EC and SMC. Experimental antisense strategies directed against RelA p65 and NF-kappaB1 p50 in early atherosclerosis and restenosis are promising in HCAEC but will be confronted with redundant pathways in HCMSMC and HCPSMC.
核因子-κB（NF-κB）的激活是早期动脉粥样硬化和再狭窄的关键事件之一。我们推测，肿瘤坏死因子-α（TNF-α）诱导并由NF-κB介导的细胞间黏附分子-1（ICAM-1）的表达可被反义RelA p65和NF-κB1 p50寡核苷酸（RelA p65和NF-κB1 p50）抑制。
成功分离（HCPSMC、HUVEC）、鉴定并培养（HCPSMC、HCMSMC、HUVEC、HCAEC）了来自人类冠状动脉斑块材料的平滑肌细胞（HCPSMC，52例患者的斑块材料）、来自人类冠状动脉中膜的平滑肌细胞（HCMSMC）、来自脐静脉的人类内皮细胞（HUVEC）和人类冠状动脉内皮细胞（HCAEC）。在TNF-α刺激（20 ng/mL，6小时）前12小时加入RelA p65和NF-κB1 p50（1、2、4、10、20和30 μM）及对照，持续18小时。在HUVEC和HCAEC中，加入RelA p65和NF-κB1 p50后，ICAM-1表达呈剂量依赖性抑制。HCMSMC与RelA p65和NF-κB1 p50孵育后未见抑制作用。同时向HCPSMC加入RelA p65和NF-κB1 p50后，发现ICAM-1表达有中度抑制，单独加入RelA p65和NF-κB1 p50后未检测到抑制作用。
数据表明，内皮细胞和平滑肌细胞之间在NF-κB介导的ICAM-1表达上存在差异。针对早期动脉粥样硬化和再狭窄中RelA p65和NF-κB1 p50的实验性反义策略在HCAEC中有前景，但在HCMSMC和HCPSMC中将面临冗余途径。
