The p38 mitogen-activated kinase pathway regulates the human interleukin-10 promoter via the activation of Sp1 transcription factor in lipopolysaccharide-stimulated human macrophages
The p38 mitogen-activated kinase pathway regulates the human interleukin-10 promoter via the activation of Sp1 transcription factor in lipopolysaccharide-stimulated human macrophages
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DOI：
10.1074/jbc.m011157200
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发表时间：
2001-04-27
影响因子：
4.8
通讯作者：
Kumar, A
中科院分区：
文献类型：
--
作者：
Ma, W;Lim, W;Kumar, A
白介素10(IL-10)是一种抑制炎症反应和细胞免疫反应的多效性细胞因子，由多种细胞类型产生，包括T、B细胞和单核/巨噬细胞。促炎和抗炎细胞因子的调节被认为涉及不同的信号通路。在本研究中，我们研究了人IL-10(hIL-10)启动子在脂多糖(LPS)激活后对人单核细胞系THP-1的调控。对hIL-10启动子序列的分析表明，位于-652和-571之间的DNA序列是IL-10转录所必需的。对碱基对-652和-571之间的启动子序列的计算机分析表明，Sp1、PEA1、YY1和Epstein-Parr病毒特异性核抗原-2(EBNA-2)样转录因子存在共同序列。而含有PEA1、YY1和EBNA-2类转录因子序列的载体不影响hIL-10的启动子活性。为了了解Sp1激活的上游事件，我们通过使用p38和细胞外信号调节的丝裂原激活的蛋白激酶的特异性抑制剂来研究它们的作用。P38特异性抑制剂SB202190和SB203580可抑制内毒素诱导的IL-10的产生。相比之下，PD98059，一种细胞外信号调节激酶的特异性抑制剂，未能调节IL-10的产生。此外，SB203580抑制了脂多糖诱导的Sp1的激活，以及在含有Sp1共同序列的质粒中的启动子活性。这些结果表明，p38丝裂原活化蛋白激酶调节脂多糖诱导的Sp1的激活，而Sp1又调节hIL-10基因的转录。
Interleukin-10 (IL-10), a pleiotropic cytokine that inhibits inflammatory and cell-mediated immune responses, is produced by a wide variety of cell types including T and B cells and monocytes/macrophages. Regulation of pro- and anti-inflammatory cytokines has been suggested to involve distinct signaling pathways. In this study, we investigated the regulation of the human IL-10 (hIL-10) promoter in the human monocytic cell line THP-1 following activation with lipopolysaccharide (LPS). Analysis of hIL-10 promoter sequences revealed that DNA sequences located between base pairs -652 and -571 are necessary for IL-10 transcription. A computer analysis of the promoter sequence between base pairs -652 and -571 revealed the existence of consensus sequences for Sp1, PEA1, YY1, and Epstein-Parr virus-specific nuclear antigen-2 (EBNA-2)-like transcription factors. THP-1 cells transfected with a plasmid containing mutant Sp1 abrogated the promoter activity, whereas plasmids containing the sequences for PEA1, YY1, and EBNA-2-like transcription factors did not influence hIL-10 promoter activity. To understand the events upstream of Sp1 activation, we investigated the role of p38 and extracellular signal-regulated kinase mitogen-activated protein kinases by using their specific inhibitors. SB202190 and SB203580, the p38-specific inhibitors, inhibited LPS-induced IL-10 production. In contrast, PD98059, a specific inhibitor of extracellular signal-regulated kinase kinases, failed to modulate IL-10 production. Furthermore, SB203580 inhibited LPS-induced activation of Sp1, as well as the promoter activity in cells transfected with a plasmid containing the Sp1 consensus sequence. These results suggest that p38 mitogen-activated protein kinase regulates LPS-induced activation of Sp1, which in turn regulates transcription of the hIL-10 gene.