The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-α and interleukin-10 production in macrophages via the cAMP-PKA-CREB pathway in a GTP-dependent manner
The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-α and interleukin-10 production in macrophages via the cAMP-PKA-CREB pathway in a GTP-dependent manner
复制标题
DOI：
10.1111/j.1365-2567.2009.03188.x
复制
发表时间：
2010-03-01
期刊：
影响因子：
6.4
通讯作者：
Zor, Tsaffrir
中科院分区：
文献类型：
--
作者：
Avni, Dorit;Philosoph, Amir;Zor, Tsaffrir
在脂多糖（LPS）刺激的巨噬细胞中，合成的磷酸神经酰胺类似物-1（PCERA-1）下调促炎细胞因子肿瘤坏死因子-α（TNF-α）的产生，并上调抗炎细胞因子白细胞介素-10（IL-10）的产生。我们以前曾报道过PCERA-1增加环磷酸腺苷（cAMP）水平。本研究的目的是描述从PCERA-1通过cAMP调节TNF-α和IL-10产生的信号通路。我们在这里表明，PCERA-1提高细胞内cAMP水平在一个鸟苷三磷酸依赖性的方式在RAW 264.7巨噬细胞。细胞可渗透的二丁酰cAMP能够模拟PCERA-1对细胞因子产生的影响，而8-氯-苯硫基-甲基腺苷-cAMP，其特异性地激活由cAMP直接激活的交换蛋白（EPAC），但不激活蛋白激酶A（PKA），未能模拟PCERA-1的活动。一致地，PKA抑制剂H89有效地阻断PCERA-1驱动的细胞因子调节以及PCERA-1刺激的cAMP反应元件结合蛋白（CREB）在Ser-133上的磷酸化。最后，PCERA-1与磷酸二酯酶（PDE）-4抑制剂咯利普兰协同作用，激活了荧光素酶报告基因的cAMP响应性转录。我们的研究结果表明，PCERA-1激活G（s）蛋白偶联受体，导致cAMP升高，cAMP通过PKA-CREB途径促进LPS刺激的巨噬细胞中TNF-α抑制和IL-10诱导。PCERA-1受体的鉴定有望为开发新型抗炎药物建立新的靶点。
P>The synthetic phospho-ceramide analogue-1 (PCERA-1) down-regulates production of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) and up-regulates production of the anti-inflammatory cytokine interleukin-10 (IL-10) in lipopolysaccharide (LPS) -stimulated macrophages. We have previously reported that PCERA-1 increases cyclic adenosine monophosphate (cAMP) levels. The objective of this study was to delineate the signalling pathway leading from PCERA-1 via cAMP to modulation of TNF-alpha and IL-10 production. We show here that PCERA-1 elevates intra-cellular cAMP level in a guanosine triphosphate-dependent manner in RAW264.7 macrophages. The cell-permeable dibutyryl cAMP was able to mimic the effects of PCERA-1 on cytokine production, whereas 8-chloro-phenylthio-methyladenosine-cAMP, which specifically activates the exchange protein directly activated by cAMP (EPAC) but not protein kinase A (PKA), failed to mimic PCERA-1 activities. Consistently, the PKA inhibitor H89 efficiently blocked PCERA-1-driven cytokine modulation as well as PCERA-1-stimulated phosphorylation of cAMP response element binding protein (CREB) on Ser-133. Finally, PCERA-1 activated cAMP-responsive transcription of a luciferase reporter, in synergism with the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results suggest that PCERA-1 activates a G(s) protein-coupled receptor, leading to elevation of cAMP, which acts via the PKA-CREB pathway to promote TNF-alpha suppression and IL-10 induction in LPS-stimulated macrophages. Identification of the PCERA-1 receptor is expected to set up a new target for development of novel anti-inflammatory drugs.