以TCF-DNA誘捕轉錄因子調升受內毒素處理的人類主動脈內皮細胞及單核細胞性白血病細胞之存活率

Increased survival of lipopolysaccharide-activated endothelial and THP-1 cells by TCF-DNA decoy

劉宛竹(Wan-tsu Liu)

指導教授 ： 黃光策

國立中正大學/工學院/化學工程學系/碩士(2013年)

摘要

敗血症患者會呈現全身性發炎、低血壓、凝血功能失衡及器官官能障礙，雖然研究指出活性蛋白質C可降低嚴重敗血症病患的死亡率由30.8%降低至24.7%，但目前對於活性蛋白質C在受內毒素感染的血管內皮及白血球影響的機制尚未完全清楚。由實驗室先前結果得知，活性蛋白質C 可以降低受內毒素感染的人類主動脈內皮細胞中的β-catenin含量，因而假設其影響細胞存活率是藉由阻斷β-catenin-TCF複合體所調控的基因表現。本研究設計cyclin D1及fra1基因調節區之TCF的鍵結序列之為誘捕 DNA用來降低細胞核內可作用的TCF含量對受內毒素感染內皮細胞及THP-1細胞(monocyte cell line)存活率及細胞釋放的細胞激素(cytokines)的影響，其結果表示誘捕轉率因子寡核酸可以提高內皮細胞及單核白血球存活率，且也降低細胞激素(cytokines)表現量。

關鍵字

細胞激素 ； 內毒素

並列摘要

Septic shock is caused mainly by lipopolysaccharide (LPS). The signaling of LPS is via Toll-like receptor 4, resulting in activation of endothelial and mononuclear cells and production inflammation cytokines such as IL-1β and TNF-α. The high mortality of septic shock is due to multiple organ failure. Activated protein C (APC) is effective to reduce mortality from 30.8％ to 24.7％ in the patient with severe sepsis appearing inflammation and organ dysfunction. The reduction of apoptosis of LPS-treated endothelium and mononuclear cells is contributed to the decreased mortality of severe sepsis by APC. However, the underlying mechanism of APC on the survival of cells remains unclear. Here, we hypothesized that the effect of APC on cell survival and pro-inflammatory cytokine production may be through blocking the β-catenin-TCF-mediated gene expression. To decrease the TCF-mediated gene expression, the cells were transfected with the TCF-4 binding sequences of cyclin D1 and fra1 using non-viral vector methods to reduce availability of TCF. We found that TCF DNA decoys increase survival of lipopolysaccharide-treated endothelial and THP-1 cells and decrease the production of TNF-α and IL-1β in THP-1 cells.

並列關鍵字

TCF

參考文獻

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