Gille J, Paxton L L, Lawley T J, Caughman S W, Swerlick R A
Department of Dermatology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
J Clin Invest. 1997 Feb 1;99(3):492-500. doi: 10.1172/JCI119184.
The regulated expression of cell adhesion molecules (CAM) on endothelial cells is central to the pathogenesis of various inflammatory processes. Retinoic acid and synthetic derivatives have been demonstrated to exert antiinflammatory effects in cutaneous diseases. To determine modes of retinoid action in the modulation of inflammatory responses, we explored effects of all-trans-retinoic acid (t-RA) on the TNFalpha-induced expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin in cultured human dermal microvascular endothelial cells. Pretreatment with t-RA specifically prevented TNFalpha-induced VCAM-1 expression, but not ICAM-1 and E-selectin induction. t-RA significantly reduced VCAM-1-dependent T cell binding to TNFalpha-treated human dermal microvascular endothelial cells as well. This differential modulation of TNFalpha-induced CAM expression by t-RA was reflected at steady state mRNA levels and in nuclear run-on studies. In transcriptional activation studies, the TNFalpha-mediated activation of the human VCAM-1 promoter was inhibited after t-RA treatment, while the ICAM-1 promoter activation was unaffected, indicating that the selective inhibition of CAM expression is regulated in part at the level of gene transcription. Furthermore, the transcriptional inhibition by t-RA appears to be mediated by its effects upon the activation of NF-kappaB-dependent complex formation. Analysis of protein-DNA binding assays revealed marked inhibition of specific NF-kappaB-dependent binding to the tandem NF-KB sites of the VCAM-1 promoter, but not to the functional NF-kappaB motif of the ICAM-1 promoter. The specific inhibition of cytokine-mediated VCAM-1 gene expression in vitro may provide a potential basis by which retinoids exert their biological effects at sites of inflammation in vivo.
内皮细胞上细胞黏附分子(CAM)的调控表达是各种炎症过程发病机制的核心。维甲酸及其合成衍生物已被证明在皮肤病中发挥抗炎作用。为了确定类维生素A在调节炎症反应中的作用方式,我们研究了全反式维甲酸(t-RA)对肿瘤坏死因子α(TNFα)诱导的人真皮微血管内皮细胞中血管细胞黏附分子-1(VCAM-1)、细胞间黏附分子-1(ICAM-1)和E-选择素表达的影响。t-RA预处理可特异性阻止TNFα诱导的VCAM-1表达,但不能阻止ICAM-1和E-选择素的诱导。t-RA也显著降低了依赖VCAM-1的T细胞与TNFα处理的人真皮微血管内皮细胞的结合。t-RA对TNFα诱导的CAM表达的这种差异调节在稳态mRNA水平和核转录实验中均有体现。在转录激活实验中,t-RA处理后TNFα介导的人VCAM-1启动子激活受到抑制,而ICAM-1启动子激活未受影响,这表明CAM表达的选择性抑制部分是在基因转录水平上调控的。此外,t-RA的转录抑制作用似乎是通过其对NF-κB依赖性复合物形成激活的影响介导的。蛋白质-DNA结合实验分析显示,VCAM-1启动子串联NF-κB位点的特异性NF-κB依赖性结合受到显著抑制,但ICAM-1启动子的功能性NF-κB基序不受影响。体外细胞因子介导的VCAM-1基因表达的特异性抑制可能为类维生素A在体内炎症部位发挥生物学作用提供潜在基础。