Silencing of Fas-associated Death Domain Protects Mice from Septic Lung Inflammation and Apoptosis
Silencing of Fas-associated Death Domain Protects Mice from Septic Lung Inflammation and Apoptosis
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DOI：
10.1164/rccm.200804-534oc
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发表时间：
2009-05-01
影响因子：
24.7
通讯作者：
Hattori, Yuichi
中科院分区：
文献类型：
--
作者：
Matsuda, Naoyuki;Yamamoto, Seiji;Hattori, Yuichi
基本原理 迫切需要更好地了解脓毒症发病机制及其导致的器官衰竭的分子机制以及新的治疗方法和靶点。越来越多的证据表明细胞凋亡在脓毒症的病理生理学中发挥着重要作用，并且细胞凋亡可能对脓毒症急性肺损伤 (ALI) 有害。 目的：我们测试了这样的假设：全身施用靶向 Fas 相关死亡结构域 (FADD) 的小干扰 RNA (siRNA)，将 procaspase-8 募集到死亡诱导信号复合物中，可能具有保护作用。 方法：通过 BALB/c 小鼠盲肠结扎穿刺 (CLP) 诱导多种微生物败血症。 CLP后10小时使用转染试剂进行siRNA的体内递送。作为阴性对照，动物接受无义（乱序）siRNA。测量和主要结果：在CLP诱导的脓毒症小鼠中，死亡受体的表面表达上调，并且FADD高表达。 DNA 片段化梯和转移酶介导的 dUTP 缺口末端标记测定表明，FADD siRNA 治疗可抑制脓毒症肺中细胞凋亡的诱导。这种 siRNA 治疗阻止了 CLIP 小鼠的 ALI 发展，血气紊乱、组织学肺损伤和肺部炎症细胞增加的结果都得到了极大改善。最后，FADD siRNA 给药显着提高了 CLP 小鼠的存活率。结论：这些结果表明死亡受体凋亡途径（包括 FADD）在脓毒症 ALI 中的病理生理学意义，以及 FADD siRNA 对于脓毒症综合征基因治疗的潜在用途。
Rationale A better understanding of the molecular mechanisms involved in the pathogenesis of sepsis and its resultant organ failure and new therapeutic approaches and targets are urgently needed. Accumulating evidence suggests that apoptosis plays an important role in the pathophysiology of sepsis and that apoptosis may be detrimental in septic acute lung injury (ALI).Objectives: We tested the hypothesis that systemic administration of small interfering RNA (siRNA) targeting Fas-associated death domain (FADD), which recruits procaspase-8 into the death-inducing signaling complex, may be protective in septic ALI and mortality.Methods: Polymicrobial sepsis was induced by cecal ligation and puncture (CLP) in BALB/c mice. In vivo delivery of siRNA was performed by using a transfection reagent at 10 hours after CLP. As a negative control, animals received nonsense (scrambled) siRNA.Measurements and Main Results: In CLP-induced septic mice, surface expression of death receptors was up-regulated, and FADD was highly expressed. DNA fragmentation ladder and transferase-mediated dUTP nick end labeling assays showed that treatment with FADD siRNA suppressed apoptosis induction in septic lungs. This siRNA treatment prevented the ALI development in CLIP mice, as indicated by the findings that blood-gas derangements, histologic lung damage, and increased pulmonary inflammatory cells were greatly improved. Finally, FADD siRNA administration dramatically improved the survival of CLP mice.Conclusions: These results indicate the pathophysiologic significance of the death receptor apoptotic pathway, including FADD, in septic ALI and the potential usefulness of FADD siRNA for gene therapy of the septic syndrome.