Therapeutic effect of in vivo transfection of transcription factor decoy to NF-κB on septic lung in mice
Therapeutic effect of in vivo transfection of transcription factor decoy to NF-κB on septic lung in mice
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DOI：
10.1152/ajplung.00164.2004
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发表时间：
2004-12-01
影响因子：
4.9
通讯作者：
Gando, S
中科院分区：
文献类型：
--
作者：
Matsuda, N;Hattori, Y;Gando, S
核因子-κ B（NF-κ B）在调节内毒素休克病理生理学中涉及的几个基因的表达中起关键作用。我们研究了在体内引入对NF-κ B结合位点具有高亲和力的合成双链DNA是否可以阻断介导肺血管渗透的基因的表达，从而为脓毒性肺衰竭提供有效的治疗。小鼠静脉注射大肠杆菌内毒素10 mg/kg，造成内毒素性休克。我们采用基因转移试剂盒，在内毒素休克后1小时体内导入NF-κ B诱饵寡核苷酸（ODN）。在10 h时，采集血样用于组胺测量和血气分析。分别通过北方和Western印迹分析测定靶分子的基因和蛋白表达水平。I-125标记的白蛋白的透肺流量被用作肺血管通透性的指标。给药内毒素引起血浆组胺和肺组织中组氨酸脱羧酶、组胺H-1受体和诱导型一氧化氮合酶的基因和蛋白表达显著增加。血气分析显示动脉PO 2、PCO 2和pH值同时降低。所有这些由内毒素诱导的事件均被转染NF-κ B诱饵ODN显著抑制，但不被其突变（乱序）形式（用作对照）抑制。我们的研究结果首次表明NF-κ B诱饵ODN在内毒素休克基因治疗中的潜在用途。
Nuclear factor-kappaB (NF-kappaB) plays a key role in regulating expression of several genes involved in the pathophysiology of endotoxic shock. We investigated whether in vivo introduction of synthetic double-stranded DNA with high affinity for the NF-kappaB binding site could block expression of genes mediating pulmonary vascular permeation and thereby provide effective therapy for septic lung failure. Endotoxic shock was induced by an intravenous injection of 10 mg/kg Escherichia coli endotoxin in mice. We introduced NF-kappaB decoy oligodeoxynucleotide (ODN) in vivo 1 h after endotoxic shock by using a gene transfer kit. At 10 h, blood samples were collected for measurement of histamine and for blood-gas analysis. Gene and protein expression levels of target molecules were determined by means of Northern and Western blot analyses, respectively. The transpulmonary flux of I-125-labeled albumin was used as an index of lung vascular permeability. Administration of endotoxin caused marked increases in plasma histamine and gene and protein expressions of histidine decarboxylase, histamine H-1 receptors, and inducible nitric oxide synthase in lung tissues. Elevated lung vascular permeability was also found. Blood-gas analysis showed concurrent decreases in arterial PO2, PCO2, and pH. All of these events induced by endotoxin were significantly inhibited by transfection of NF-kappaB decoy ODN but not by its mutated (scrambled) form (used as a control). Our results indicate for the first time the potential usefulness of NF-kappaB decoy ODN for gene therapy of endotoxic shock.