Dynamic changes of RNA and protein synthesis by neutrophils during the course of acute inflammation in rabbits.

家兔急性炎症过程中中性粒细胞RNA和蛋白质合成的动态变化。

基本信息

批准号：
03670177
负责人：
OHKAWARA Susumu
金额：
$ 1.28万
依托单位：
KUMAMOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

Neutrophils have long been considered to be terminally differentiated end cells and to possess little RNA and protein synthesis capability. Using a casein-induced rabbit peritonitis, we found that neutrophils synthesize interleukin 1beta(IL-1beta)de novo only at early stages of inflammation. This observation suggested that protein synthesis of neutrophils is involved with inflammatory reaction. With the focus on RNA/protein synthesis capability by neutrophils and its dynamic change during the inflammatory process, we carried out gene library subtraction studies. We performed gene subtraction with the ss cDNA library hybridization method and the resulted clones of 5 hr-minus 24 hr-neutrophils were further confirmed by differential hybridization. The cDNA from candidate clones were converted to sense- and anti-sense-RNA and using these RNA probes, we performed Northern blot and cytoplasmic slot blot analyses.Immune activation gene 2(Act-2), MIF related protein-8(MRP-8), MRP-14, fMLP rece … More ptor (fMLPR) and gamma-actin homologues were isolated as genes representing neutrophils at an early inflammatory stage. Ferritin light(L)chain cDNA was cloned as the gene representing neutrophils at a late inflammatory stage. In addition, mRNA expression of rabbit neutrophil attractant/activation protein-1/interleukin 8(NAP-1/IL-8), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor alpha(TNFalpha)and ferritin heavy(H)chain mRNASs expression during inflammation were examined by cytoplasmic dot blot analysis. The patterns of mRNAs of Act-2, NAP-1/IL-8, MRP-14 and gamma-actin homologues were induced immediately after the onset of inflammation. This pattern is similar to that of IL-1beta mRNA expression. Pattern2 : mRNA expression of the ferritin H chain in neutrophils reached a peak at intermediate periods (12hr)of inflammation, with a monophagic pattern. Pattern3 : the level of ferritin L chain mRNA increased with progression of the inflammatory process. The mRNA expressions of TNFalpha and MCP-1 was not significant. These data suggest that neutrophils contribute to the defense reactions by synthesizing various proteins at the site of inflammation. Even in a later stage of inflammation, neutrophils were viable and retain protein synthetic capability, although they are generally believed to have a very short life-span and may die at a different site of inflammation. Less

长期以来，嗜中性粒细胞被认为是末端分化的末端细胞，几乎没有RNA和蛋白质合成能力。使用病例诱导的兔腹膜炎，我们发现中性粒细胞合成白介素1beta（IL-1Beta）仅在炎症的早期阶段。该观察结果表明，中性粒细胞的蛋白质合成与炎症反应有关。伴随着嗜中性粒细胞的RNA/蛋白质合成能力及其在炎症过程中的动态变化，我们进行了基因文库减法研究。我们使用SS cDNA文库杂交方法进行了基因减法，并通过差异杂交进一步证实了5小时24 hr-中性基质的克隆。将来自候选克隆的cDNA转化为感官和抗渗透性RNA，并使用这些RNA问题，我们进行了北印迹和细胞质插槽斑点分析。免疫激活基因2（ACT-2），MIF相关蛋白质-8（MRP-8），MRP-14，MRP-14，MRP-14，MRP-14，FMLP CORES INSTING ISS INSTING ISLACTINIS ISLACTING IN sIMATICTING INTIC ISLACTINGIN（FMLAN-ways）（fmla toct）（fmla）（fmla）（fma）中性粒细胞在早期炎症阶段。铁蛋白光（L）链cDNA被克隆为在炎症阶段代表中性粒细胞的基因。 In addition, mRNA expression of rabbit neutrophil attractant/activation protein-1/interleukin 8(NAP-1/IL-8), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor alpha(TNFalpha) and ferritin heavy(H)chain mRNAS expression during inflammation were examined by cytoplasmic dot blot analysis.炎症发作后，立即诱导了ACT-2，NAP-1/IL-8，MRP-14和γ-肌动蛋白同源物的mRNA的模式。该模式类似于IL-1Beta mRNA表达的模式。模式2：中性粒细胞中铁蛋白H链的mRNA表达在炎症的中间时期（12hr）达到峰值，具有单性模式。模式3：铁蛋白L链mRNA的水平随炎症过程的进展而增加。 TNFALPHA和MCP-1的mRNA表达不显着。这些数据表明，中性粒细胞通过在炎症部位合成各种蛋白质来促进防御反应。即使在炎症的后期，中性粒细胞也是可行的，并且保留了蛋白质合成能力，尽管通常认为它们的寿命很短，并且可能在不同的炎症部位死亡。较少的