The transcription factor AP-1 as a molecular target in sepsis therapy

转录因子 AP-1 作为脓毒症治疗的分子靶点

基本信息

批准号：
20590250
负责人：
HATTORI Yuichi
金额：
$ 3.08万
依托单位：
University of Toyama
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

项目摘要

Polymicrobial sepsis was induced by cecal ligation and puncture (CLP) in BALB/c mice (8-12 wk of age). The DNA binding activity of AP-1, as assessed by electrophoretic mobility shift assay, was time-dependently increased in lung tissues from mice after the onset of CLP-induced sepsis. This increase was effectively eliminated by in vivo transfection of AP-1 decoy oligonucleotides (ODNs). Sepsis induction significantly increased surface expression of death receptors, such as TNF-R1, Fas, DR4, and DR5, in lung and aortic tissues after sepsis. Furthermore, the gene and protein levels of FADD, which recruits procaspase-8 into the death-inducing signaling complex, were increased after sepsis induction. These sepsis-induced changes were eliminated by systemic application of AP-1 decoy ODNs. TUNEL assays revealed that the significant appearance of cell apoptosis in lung and aortic tissue sections from septic mice was prevented by systemic treatment with AP-1 decoy ODNs. When endotoxic shock was induced by an intravenous injection of 10mg/kg lipopolysaccharide (LPS) in mice, expression levels of inflammatory molecules, including IL-1R, IL-6R, HMGB-1, and gp130, were highly increased, which was significantly inhibited by AP-1 decoy ODN treatment. All animals which received LPS died within 48h, and the animals that were treated with AP-1 decoy ODNs after LPS exhibited a striking improvement of survival. Our results suggest that AP-1 decoy ODN therapy represent an effective strategy in the treatment of sepsis. In addition, systemic administration of siRNA targeting FADD, which was found to be transactivated by AP-1, prevented the development of acute lung injury in CLP mice, and improved their survival. These findings indicate the potential usefulness of FADD siRNA for gene therapy of the septic syndrome.

BALB/c 小鼠（8-12 周龄）通过盲肠结扎穿刺 (CLP) 诱导多种微生物败血症。通过电泳迁移率变动测定评估，在 CLP 诱导的脓毒症发作后，小鼠肺组织中 AP-1 的 DNA 结合活性呈时间依赖性增加。通过体内转染 AP-1 诱饵寡核苷酸 (ODN) 可以有效消除这种增加。脓毒症诱导显着增加了脓毒症后肺和主动脉组织中死亡受体的表面表达，例如 TNF-R1、Fas、DR4 和 DR5。此外，FADD（将 procaspase-8 招募到死亡诱导信号复合物中）的基因和蛋白质水平在脓毒症诱导后增加。这些脓毒症引起的变化通过全身应用 AP-1 诱饵 ODN 得以消除。 TUNEL 分析显示，脓毒症小鼠肺和主动脉组织切片中细胞凋亡的显着出现可以通过 AP-1 诱饵 ODN 的全身治疗来预防。小鼠静脉注射10mg/kg脂多糖（LPS）诱导内毒素休克时，IL-1R、IL-6R、HMGB-1、gp130等炎症分子表达水平大幅升高，而炎症分子表达水平明显受到抑制通过AP-1 decoy ODN处理。所有接受LPS的动物均在48小时内死亡，并且在LPS后用AP-1诱饵ODN处理的动物表现出显着的生存改善。我们的结果表明 AP-1 诱饵 ODN 疗法是治疗脓毒症的有效策略。此外，全身注射针对 FADD（被发现可被 AP-1 反式激活）的 siRNA，可以防止 CLP 小鼠发生急性肺损伤，并提高其存活率。这些发现表明 FADD siRNA 对于脓毒症综合征基因治疗的潜在用途。