Three Arachidonoylamide Derivatives Inhibit Pro-Inflammatory Genes Expression by Modulating NF-κB and AP1 Activities
Three Arachidonoylamide Derivatives Inhibit Pro-Inflammatory Genes Expression by Modulating NF-κB and AP1 Activities
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DOI：
10.2174/1573406412666160502154936
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发表时间：
2016-01-01
影响因子：
2.3
通讯作者：
Menegazzi, Marta
中科院分区：
文献类型：
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作者：
Gregorelli, Alex;Sgarbossa, Anna;Menegazzi, Marta
背景：由于花生四烯酸衍生物的抗炎活性已有报道，我们合成了三种新的花生四烯酸酰胺衍生物（AA-Ds），并在体外皮肤炎症模型上测试了它们的抗炎作用。我们的研究目的是寻找能够下调炎症信号转导途径的天然化合物衍生物。方法：在AA-Ds存在的情况下，采用细胞因子诱导培养人角化细胞（HaCaT）。细胞因子管理引起NF-kappa B和STAT-1激活介导的炎症反应，诱导促炎基因表达。结果：通过实时荧光定量PCR发现，诱导24小时后，所有AA-Ds均能显著抑制诱导型一氧化氮合酶（iNOS）、TNF α、nf - κ B抑制剂α、趋化因子（C-X-C基序）配体9和10个基因的表达。我们分析了它们对iNOS基因表达的分子效应。由于AA-Ds处理没有改变iNOS转录半衰期，因此我们排除了该基因转录后调控的突出作用，并将注意力集中在其转录调控上。从细胞因子诱导后3 - 5小时开始，用每种化合物预处理的HaCaT细胞显示NF-kappa B dna结合和NF-kappa B p65-Ser536磷酸化均受到抑制。只有AA-D4衍生物能抑制STAT1的激活。为了解释为什么诱导后iNOS表达的抑制开始较晚，我们分析了其他关键转录因子的活性。AA-Ds处理引起AP1 DNA结合以及c-Jun、c-Fos和Fra-1 mRNA水平的早期升高。我们的数据与先前在其他细胞系统中描述的AP1对人类iNOS启动子的抑制作用一致（Kleinert et al.）。结论：AA-Ds是几种促炎基因诱导的良好候选抑制剂，本研究为其作为新型抗炎药物提供了适应症。
Background: Since the anti-inflammatory activity of arachidonic acid derivatives was previously reported, we synthesized three new amide derivatives of arachidonic acid (AA-Ds) and tested their anti-inflammatory effects on an in vitro skin inflammation model. Aim of our study was to find derivatives of natural compounds able to down regulate inflammatory signal transduction pathway.Methods: Human keratinocytes cell line (HaCaT) was cultured and induced by cytokines in the presence of AA-Ds. Cytokines administration elicited an inflammatory response mediated by NF-kappa B and STAT-1 activation that induced proinflammatory genes expression.Results: By real time PCR we found that 24 hours after induction all AA-Ds significantly inhibit inducible Nitric Oxide Synthase (iNOS), TNF alpha, Inhibitor alpha of NF-kappa B, chemokine (C-X-C motif) ligand 9 and 10 genes expression. We analyzed their molecular effects in particular on the iNOS gene expression. Since iNOS transcript half-life did not change with AA-Ds treatment, we excluded a prominent role of post-transcriptional regulation for this gene and focused our attention on its transcriptional regulation. Starting three-five hours after cytokines induction, HaCaT cells, pre-treated with each compound, showed inhibition of both NF-kappa B DNA-binding and NF-kappa B p65-Ser536 phosphorylation. STAT1 activation was inhibited only by AA-D4 derivative. To explain why the inhibition of iNOS expression began late after induction we analyzed activities of others key transcription factors. AA-Ds treatment elicited early increases of AP1 DNA binding as well as c-Jun, c-Fos and Fra-1 mRNA levels. Our data agree with the repressing effects of AP1 on human iNOS promoter previously described in others cell systems (Kleinert et al.).Conclusion: AA-Ds shown to be good candidates as inhibitors of several pro-inflammatory genes induction and our study provides indications for their possible use as new antiinflammatory drugs.