NF-kappaB对ICAM-1表达的调节作用

Role of NF-kappa B in regulating expression of ICAM-1

目的:研究NF-kappaB对粘附分子ICAM-1表达的调控作用。方法:原代培养HUVECs,用LPS(100 ng.ml-1)刺激不同时间(1、4、8、24 h)后,其中于4 h处预先用NF-κB抑制剂PDTC(1 mmol.L-1)处理15 min,再分别提取核蛋白进行凝胶电泳迁移改变分析(Electrophoretical Mobility Shift Assay EMSA)法测定NF-kappaB活性及提取总RNA进行RT-PCR检测ICAM-1mRNA表达。结果:NF-kappaB的活性变化与ICAM-1的表达有着明显的一致性。用PDTC(NF-kappaB抑制剂)处理内皮细胞,发现ICAM-1的表达随着NF-kappaB活性的降低而下调(2.40±0.11vs0.76±0.09,P<0.01)。结论:NF-kappaB对ICAM-1基因表达起着重要的调控作用。

AIM： To study the role of NF-kappa B in regulating the expression of adhesion molecule ICAM-1. METHODS： HUVECs were isolated from human umbilical veins by 0.25% trypsin and then cultured in RPMI1640, exposed on LPS （100 ng·ml^-1） for different time （ 1, 4, 8, and 24 h）, while pretreated by NF-kappa B inhibitor pyrrolidine dithiocarbamate （PDTC） at the 4th hour. Nucleus protein and total RNA were extracted. The activity of NF-kappa B and the expression of ICAM-1 were measured respectively by the electrophoretic mobility shift assay （EMSA） and reverse transcriptase-PCR （RT- PCR）. RESULTS： The activity of NF-kappaB altered in correlate with ICAM-1. Treating HUVECs with PDTC,the expression of ICAM-lmRNA was significantly down- regulated with the decline activity of NF-kappa B （2.40 ±0.11 vs 0.76±0.09, P 〈0.01）. CONCLUSION： NF-kappa B plays an important role in regulating the expression of ICAM- 1Mrna.