Van Kampen C, Mallard B A
Department of Pathobiology, University of Guelph, Ont., N1G 2W1, Guelph, Canada.
Vet Immunol Immunopathol. 2001 May 10;79(1-2):129-38. doi: 10.1016/s0165-2427(01)00251-3.
Endothelial cell adhesion molecules (AM) intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) are important mediators of cell migration from blood into tissue. The kinetics of ICAM-1 mRNA and VCAM-1 protein expression in bovine aortic endothelial cells (BAEC) were determined using quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) and flow cytometric analysis, respectively. Stimulation of BAEC with recombinant bovine tumor necrosis factor alpha (rbTNF-alpha) resulted in protein expression of VCAM-1 on less than 5% of all cultured BAECs at 1h post-stimulation, followed by a significant increase at 3h that was maintained until 48h when the proportion of VCAM-1 positive (+) cells decreased significantly, but not to baseline proportions. The expression kinetics for VCAM-1 were similar on cells stimulated with lipopolysaccharide (LPS) except at 24h, when there was a significantly higher proportion of BAEC expressing VCAM-1 than at any other time. The expression of ICAM-1 mRNA differed significantly between stimuli. Expression of ICAM-1 mRNA peaked at 12-18h and then diminished but remained at amounts above baseline up to 72h after stimulation. Stimulation with LPS induced a significant increase in ICAM-1 mRNA expression between 1 and 12h after which the amounts rapidly decreased to baseline. In summary, different stimuli produced similar expression kinetics of VCAM-1 surface protein but different kinetics of ICAM-1 mRNA expression.
内皮细胞黏附分子(AM)细胞间黏附分子1(ICAM - 1)和血管细胞黏附分子1(VCAM - 1)是细胞从血液迁移至组织的重要介质。分别使用定量逆转录聚合酶链反应(Q - RT - PCR)和流式细胞术分析,测定了牛主动脉内皮细胞(BAEC)中ICAM - 1 mRNA和VCAM - 1蛋白的表达动力学。用重组牛肿瘤坏死因子α(rbTNF - α)刺激BAEC后,在刺激后1小时,所有培养的BAEC中不到5%的细胞表达VCAM - 1蛋白,随后在3小时显著增加,并持续至48小时,此时VCAM - 1阳性(+)细胞比例显著下降,但未降至基线比例。用脂多糖(LPS)刺激的细胞上VCAM - 1的表达动力学相似,除了在24小时时,表达VCAM - 1的BAEC比例显著高于其他任何时间。不同刺激下ICAM - 1 mRNA的表达差异显著。ICAM - 1 mRNA的表达在12 - 18小时达到峰值,然后下降,但在刺激后72小时内仍高于基线水平。用LPS刺激在1至12小时之间诱导ICAM - 1 mRNA表达显著增加,之后其含量迅速降至基线。总之,不同刺激产生了相似的VCAM - 1表面蛋白表达动力学,但ICAM - 1 mRNA表达动力学不同。