Gene expression profile of antithrombotic protein C defines new mechanisms modulating inflammation and apoptosis
Gene expression profile of antithrombotic protein C defines new mechanisms modulating inflammation and apoptosis
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DOI：
10.1074/jbc.c100017200
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发表时间：
2001-04-06
影响因子：
4.8
通讯作者：
Grinnell, BW
中科院分区：
文献类型：
--
作者：
Joyce, DE;Gelbert, L;Grinnell, BW
人蛋白 C 是一种天然抗凝因子，该分子的重组激活形式 (rhAPC) 正在完成治疗严重脓毒症的临床评估。由于内皮功能障碍在严重炎症性疾病和脓毒症中的病理生理学作用，我们探索了rhAPC可能直接调节内皮功能的可能性，而不依赖于其抗凝活性。通过广泛的转录分析，我们发现 rhAPC 直接调节内皮细胞基因表达模式，聚类成抗炎和细胞存活途径，rhAPC 直接抑制 p50 和 p52 NF kappaB 亚基的表达，导致靶位点 NF kappaB 结合的功能性降低。此外，rhAPC 阻断肿瘤坏死因子 α 诱导后下游 NF kappaB 调节基因的表达，包括剂量依赖性抑制细胞粘附表达以及细胞内粘附分子 1、血管细胞粘附分子 1 和 E-选择素的功能性结合。此外，rhAPC 调节内皮细胞凋亡途径中的多个基因，包括 Bcl-2 同源蛋白和凋亡蛋白抑制剂。这些途径的变化导致 rhAPC 能够抑制强效诱导剂星形孢菌素诱导的细胞凋亡。这种对内皮调节和肿瘤坏死因子诱导的内皮功能障碍的调节的新机制理解在凝血、炎症和细胞死亡之间建立了新的联系，并为 APC 在全身炎症和脓毒症中的功效提供了分子基础的见解。
Human protein C is a natural anticoagulant factor, and a recombinant activated form of the molecule (rhAPC) is completing clinical evaluation for treatment of severe sepsis. Because of the pathophysiologic role of endothelial dysfunction in severe inflammatory disease and sepsis, we explored the possibility that rhAPC might directly modulate endothelial function, independent of its anticoagulant activity. Using broad transcriptional profiling, we show that rhAPC directly modulates patterns of endothelial cell gene expression clustering into anti-inflammatory and cell survival pathways, rhAPC directly suppressed expression of p50 and p52 NF kappaB subunits, resulting in a functional decrease in NF kappaB binding at target sites. Further, rhAPC blocked expression of downstream NF kappaB regulated genes following tumor necrosis factor alpha induction, including dose-dependent suppression of cell adhesion expression and functional binding of intracellular adhesion molecule 1, vascular cell adhesion molecule 1, and E-selectin. Further, rhAPC modulated several genes in the endothelial apoptosis pathway, including the Bcl-2 homologue protein and inhibitor of apoptosis protein. These pathway changes resulted in the ability of rhAPC to inhibit the induction of apoptosis by the potent inducer, staurosporine. This new mechanistic understanding of endothelial regulation and the modulation of tumor necrosis factor-induced endothelial dysfunction creates a novel link between coagulation, inflammation, and cell death and provides insight into the molecular basis for the efficacy of APC in systemic inflammation and sepsis.