Evaluation of Gs protein regulation in septic heart of gene deficiency mice

基因缺陷小鼠脓毒症心脏中 Gs 蛋白调控的评价

基本信息

批准号：
13470316
负责人：
GANDO Satoshi
金额：
$ 4.1万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470316/
关键词：
sepsis heart Gs-α nuclear factor-κB gene therapy knockout mice 心収縮性 Gsα蛋白遺伝子欠損動物 Toll-like受容体 macrophage migration inhibitory factor nuclear factor κB G蛋白 TNF-a 敗血症性ショック心筋陽性変力作用盲腸結紮モデル細胞内情報伝達

项目摘要

Sepsis can cause a direct change of intracellular signaling in the heart, which includes reduction of positive inotropic effects by diminished Gsα proteins. The decrease of Gsα was investigated in this study by using gene targeting therapy and caecum legation and puncture (CLP) and injection of lipopolysaccharide (LPS).Cardiac Gsα was decreased to about 45% from 36 h by death time in the CLP mice. Anti TNF-α antibody, anti IL-1β antibody, anti IL-6 antibody and iNOS selective inhibitor, FR26U330, cannot reverse the decrease of Gsα, in CLP. Some knockout mice of inflammatory cytokine, such as iNOS, showed no effects of the regulation of Gsα in sepsis. As a result of CLP mice, gene therapies of nuclear factor-xB (NF-κB) were easily uptaken in the lung and the atrium and were able to inhibit the production of inflammatory cytokines, such as iNOS. However, a gene therapy of nuclear factor -κB (NF-κB) cannot improve Gsα reduction in the heart of CLP mice. Even in septic rabbits by injection of LPS, the, diminished amounts of Gsα in the atrium were not significantly improved by NF-κB gene therapy. In this study, the regulation of Gsα and intracelluer signaling via histamine remained to be diminished in septic atrium.Imrnunohistochemistry revealed that the expression of Toll-like receptor 4 (TLR4) appeared in the atrium, which can induce inflammatory cytokines by NF-κB activation in gel-mobility shift assay and northern blot analysis. Knockout mice of macrophage migration inhibitory factor (MTF) inhibited TLR4 expression in the atrium and significantly improved the depression of the amount of Gsα in Western blot analysis.We conclude that sepsis cannot reduce Gsα in the heart by activation of transcriptional factor NF-κB but MAP kinase via stimulation of TLR4.

败血症会导致心脏中细胞内信号的直接变化，其中包括减少GSα蛋白的阳性肌力作用。在这项研究中，通过使用基因靶向治疗，CAECUM CRIGATION和PUNCTURE（CLP）和注射脂多糖（LPS）来研究GSα的降低。从CLP小鼠死亡时，降低了脂多糖（LPS）。抗TNF-α抗体，抗IL-1β抗体，抗IL-6抗体和iNOS选择性抑制剂FR26U330不能反转CLP中GSα的降低。一些炎症细胞因子的敲除小鼠，例如iNOS，显示败血症中GSα的调节没有影响。由于CLP小鼠，核因子-XB（NF-κB）的基因疗法很容易在肺和中庭中吸收，并能够抑制炎性细胞因子（例如Inos）的产生。但是，核因子-κB（NF -κB）的基因疗法不能改善CLP小鼠心脏的GSα降低。即使通过注射LPS的化粪池兔子，NF-κB基因疗法也无法显着改善中庭的GSα量减少。在这项研究中，通过组胺对GSα和细胞内信号的调节仍有望减少。IMRNUNOO组织表明，在中庭中出现了类似TOLL样受体4（TLR4）的表达，可以通过NF-κB激活性诱导炎症性细胞因子在凝胶型转移中诱导炎症性细胞因子，并分析凝胶型分析。巨噬细胞迁移抑制因子（MTF）的敲除小鼠抑制了中庭中TLR4的表达，并显着改善了蛋白质印迹分析中GSα量的抑郁症。我们包括，败血症不能通过激活转录因子NF-κB来降低心脏中的GSα，但是通过刺激TLR4的刺激来激活NF-κB。