Molecular analysis of cyclooxigenase gene expression and its application for new designs of anti-inflammatory drug

环加氧酶基因表达的分子分析及其在抗炎药物新设计中的应用

基本信息

批准号：
11670450
负责人：
OTSUKA Takeshi
金额：
$ 1.86万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

(monocytes/macrophages)CD40 expression of human peripheral blood monocytes (PBMs) was enhanced in the presence of IFN-g, and further induction was achieved by GM-CSF.So far, signaling cascades of CD40 have been examined by using the cells expressing CD40L on their cell membrane. We used anti CD40 antibody as a stimulant, and it was strong enough to induce PBMNs productions of both TNFa and PGE2. These results indicated that CD40 signaling does not need an interaction of other costimulatory molecules. Induced PGE2 production was accompanied by the induction of COX-2 on the level of gene expression. Both MAP Kinases and NFkBwere activated by CD40L-CD40 interaction, although their activation profiles were different from those stimulated by LPS.(neutrophils)In LPS-stimulated human neutrophils, the activation of ERK and p38 MAPK is independently involved in the induction of COX-2 protein, and in PGE2 synthesis. LPS induces the phosphorylation and activation of ERK2 and p38 MAPK in neutrophi … More ls.Both IL-4 and IL-10 completely inhibited LPS-induced COX-2 expression and PGE2 synthesis in neutrophils. Neither of these cytokines alone induced the phosphorylation or activation of these Kinases. Interestingly, IL-10 partly inhibited LPS-induced phosphorylation and activation of p38 MAPK, although IL-4 did not affect LPS-induced phosphorylation or activation of p38 MAPK.In addition, LPS-induced phosphorylation and activation of ERK2 was not affected at all by these cytokines. These results suggest that LPS-induced phosphorylation and activation of p38 MAPK was partly regulated by IL-10, but not by IL-4.Activation of peripheral blood monocytes and neutrophils of patients with rheumatoid arthritis (RA) should be different from those stimulated with LPS in vitro. PGE2 production by RA neutrophils was higher than that by normal neutrophils, and it was chiefly carried out by cyclooxygenase (COX)-2. By using inhibitors for MAP kinase cascade, RA neutrophils showed the activation both of ERK2 and p38 MAPK.IL-4, IL-10 or dexamethasone were able to suppress LPS-induced but not constitutively elevated PGE2 production in RA neutrophils. These results addressed the question that activated condition of RA neutrophils is different from that of neutrophils stimulated by LPS. Less

（单核细胞/巨噬细胞）在IFN-G存在下增强了人外周血单核细胞（PBM）的CD40表达，并且通过在其细胞膜上使用表达CD40L的细胞来检查CD40的信号级联的CD40的信号级联。我们使用抗CD40抗体作为刺激剂，并且足以诱导TNFA和PGE2的PBMNS生产。这些结果表明，CD40信号传导不需要其他共刺激分子的相互作用。通过在基因表达水平上引入COX-2来实现诱导的PGE2产生。 MAP激酶和NFKBWERE都被CD40L-CD40相互作用激活，尽管它们的激活曲线与LPS刺激的激活曲线不同。（中性粒细胞）在LPS刺激的人类嗜中性粒细胞中，ERK和p38 MAPK的激活与Cox-2蛋白质和PGE2综合诱导诱导了Cox-2蛋白质。 LPS在中性粒细胞中诱导ERK2和p38 MAPK的磷酸化和激活……更多的LSS IL-4和IL-10完全抑制了LPS LPS诱导的Cox-COX-COX-COX-COX-COX-2的表达和PGE2中性粒细胞中的PGE2合成。这些细胞因子都没有诱导这些激酶的磷酸化或激活。有趣的是，尽管IL-4不影响LPS诱导的磷酸化或p38 MAPK的激活，但IL-10部分抑制了LPS诱导的p38 MAPK的磷酸化和激活。这些结果表明，LPS诱导的p38 MAPK的磷酸化和激活部分受IL-10的调节，但不通过IL-10调节。4。类风湿关节炎患者的外周血单核细胞和中性粒细胞的激活应与LPS在VITO中刺激的LPS的患者（RA）不同。 RA嗜中性粒细胞产生的PGE2高于正常嗜中性粒细胞的PGE2，首先是通过环氧合酶（COX）-2进行的。通过将抑制剂用于MAP激酶级联反应，RA中性粒细胞显示ERK2和p38 MAPK.IL-4，IL-10或地塞米松都能够抑制LPS诱导的RA中性粒细胞中LPS诱导的PGE2产生。这些结果解决了一个问题，即在LPS刺激的嗜中性粒细胞的RA中性粒细胞的激活状况不同。较少的