Lauth M, Wagner A H, Cattaruzza M, Orzechowski H D, Paul M, Hecker M
Department of Cardiovascular Physiology, University of Göttingen, Germany.
J Mol Med (Berl). 2000;78(8):441-50. doi: 10.1007/s001090000129.
Deformation-induced synthesis of endothelin-1 (ET-1) in endothelial cells exposed to high blood pressure may play an important role in vein graft disease and in restenosis following percutaneous transluminal angioplasty. Effective inhibitors of preproendothelin ET-1 (ppET-1) processing to ET-1 are not available, and blockade of ppET-1 expression may therefore emerge as an alternative therapeutic approach. To evaluate this, we investigated deformation-sensitive transcription factors controlling ppET-1 expression in both native (rabbit carotid artery and jugular vein) and cultured endothelial cells (EC; porcine aorta and human umbilical vein). Deformation of both native and cultured endothelial cells for 6 h resulted in a marked increase in ET-1 synthesis which was preceded by a transient (30-60 min) activation of transcription factors activator protein-1 (AP-1) and CCAAT/enhancer-binding protein (C/EBP) beta and/or delta. A decoy oligodeoxynucleotide directed against AP-1 inhibited deformation-induced ppET-1 expression in the rabbit jugular vein as well as in porcine aorta EC and human umbilical vein EC but not in the rabbit carotid artery. Subsequent reporter gene analyses with different rat ppET-1 promoter-luciferase constructs transiently transfected into porcine aorta EC identified a single AP-1 binding site at -110 to -100 bp as the primary response element for deformation-induced ppET-1 expression. Moreover, a C/EBP-specific decoy oligodeoxynucleotide abolished ppET-1 expression in the endothelium of the rabbit carotid artery, but not in the jugular vein where basal ET-1 synthesis was greatly enhanced instead. These findings suggest that the key transcription factors controlling deformation-induced ppET-1 expression in endothelial cells are blood vessel rather than species-specific. In humans, adjunct treatment with an AP-1-specific decoy oligodeoxynucleotide may prove be an interesting gene therapeutic option for the above cardiovascular interventions.
在暴露于高血压的内皮细胞中，变形诱导的内皮素 -1（ET-1）合成可能在静脉移植物疾病以及经皮腔内血管成形术后再狭窄中起重要作用。目前尚无有效的前内皮素原ET-1（ppET-1）加工成ET-1的抑制剂，因此阻断ppET-1表达可能成为一种替代治疗方法。为评估这一点，我们研究了在天然（兔颈动脉和颈静脉）和培养的内皮细胞（EC；猪主动脉和人脐静脉）中控制ppET-1表达的变形敏感转录因子。天然和培养的内皮细胞变形6小时导致ET-1合成显著增加，这之前转录因子激活蛋白-1（AP-1）和CCAAT/增强子结合蛋白（C/EBP）β和/或δ短暂（30 - 60分钟）激活。针对AP-1的诱饵寡脱氧核苷酸抑制了兔颈静脉以及猪主动脉EC和人脐静脉EC中变形诱导的ppET-1表达，但在兔颈动脉中未起作用。随后用不同的大鼠ppET-1启动子 - 荧光素酶构建体瞬时转染猪主动脉EC进行报告基因分析，确定在 -110至 -100 bp处的单个AP-1结合位点是变形诱导的ppET-1表达的主要反应元件。此外，C/EBP特异性诱饵寡脱氧核苷酸消除了兔颈动脉内皮中的ppET-1表达，但在颈静脉中未起作用，反而基础ET-1合成大大增强。这些发现表明，控制内皮细胞中变形诱导的ppET-1表达的关键转录因子是血管特异性而非物种特异性。在人类中，用AP-1特异性诱饵寡脱氧核苷酸进行辅助治疗可能被证明是上述心血管干预的一种有趣的基因治疗选择。
