利用TCF誘捕DNA提昇內毒素感染之人類主動脈內皮細胞Cyclic AMP之相關訊號傳遞

敗血症被定義為因感染所造成的全身性發炎反應綜合症群(systemic inflammatory response syndrome；SIRS)，嚴重的敗血症患者會產生急性呼吸窘迫症(respiratory distress syndrome；ARDS)、多重器官官能障礙症候群(multiple organ failure；MOF)等因而導致死亡。在臨床研究上，活性蛋白質C(Active protein C；APC)用於治療嚴重性敗血症患者能減少百分之6.1的死亡率，然而雖已被證實具有療效，但關於其於受內毒素(lipopolysaccharide；LPS)感染的內皮細胞(endothelial cells；ECs)藥理機制尚未完全清楚。由先前實驗室結果得知，活性蛋白質C能降低內皮細胞質中β-catenine的含量，因此我們使用非病毒式載體轉染TCF誘捕DNA (T cell factor DNA decoy)。並假設細胞內的Cyclic AMP (cAMP)會因為TCF誘捕DNA阻止細胞週期相關的cylin D1基因轉錄進而增加。實驗結果得知，TCF誘捕DNA可抑制受內毒素感染的內皮細胞凋亡，其藥效的作用之一是提升LPS感染的細胞內cAMP濃度。

關鍵字

環腺苷酸；內毒素； TCF誘捕DNA

並列摘要

Sepsis is defined as being a systemic inflammatory response syndrome (SIRS). The critically ill patients appear acute respiratory distress syndrome (ARDS), and multiple organ failure/dysfunction syndrome (MOF/MODS); and these developments remain the leading cause of death. In a previous study, the administration of activated protein C (APC) reduced the rate of death in patients with a clinical diagnosis of severe sepsis by 6.1%. Although APC is effective to reduce the mortality in sepsis patients, the underlying mechanism of APC on the survival of lipopolysaccharide-treated endothelial cells (ECs) remains unclear. In our previous study, we found that APC is able to reduce the level of cytosolic β-catenin. Therefore we transfected the cells with several DNA decoys derived from the T cell factor (TCF) binding sequences using non-viral vector methods. We hypothesized that the effect of APC on cell survival may be due to blocking the β-catenin-TCF-mediated gene expression including Cyclin D1 which was related to cell cycle. We also hypothesized that the level of cyclic-AMP (cAMP) might be increased due to the inhibition of cell cycle by the TCF decoy. Our study found that TCF DNA decoys increased the intracellular cAMP production. This would activate cAMP response-element binding protein (CREB) and finally negatively regulate expression of NF-κB. These results demonstrated the therapeutic potential of TCF DNA decoys on treatment of severe septic shock and would bypass the side effect of APC on bleeding.

並列關鍵字

LPS ； TCF DNA decoy ； Cyclic AMP

參考文獻

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[2] Hattori Y, Takano K, Teramae H, Yamamoto S, Yokoo H, Matsuda N. "Insights into sepsis therapeutic design based on the apoptotic death pathway". J Pharmacol Sci. 2010,114(4):354-65.

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國際替代計量

利用TCF誘捕DNA提昇內毒素感染之人類主動脈內皮細胞Cyclic AMP之相關訊號傳遞

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