Erkel G, Wisser G, Anke T
Department of Biotechnology, University of Kaiserslautern, Paul-Ehrlich-Str. 23, D-67663 Kaiserslautern, Germany.
Int Immunopharmacol. 2007 May;7(5):612-24. doi: 10.1016/j.intimp.2007.01.001. Epub 2007 Jan 29.
The fungal secondary metabolite panepoxydone has been recently described as an inhibitor of NF-kappaB activation which is a pivotal regulator of the inflammatory and immune response. These findings have led to propose that panepoxydone may be useful as anti-inflammatory agent. In this study we investigated for the first time the effects of panepoxydone on inflammatory gene expression in the monocytic cell line MonoMac6, stimulated with lipopolysaccharide (LPS) and the phorbolester 12-O-tetradecanoylphorbol-13-acetate (TPA). DNA microarray analysis of 110 human genes known to be strongly regulated during inflammation, combined with reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) revealed that low micromolar concentrations (12-24 microM) of panepoxydone strongly inhibited the expression of thirty-three NF-kappaB dependent pro-inflammatory genes such as the chemokines CCL3, CCL4, CCL8; CXCL8, CXCL10, CXCL20, the cytokines IL-1, IL-6, TNF-alpha, pro-inflammatory enzymes like COX-2, and components of the REL/NF-kappaB/IkappaB family without significant effects on the expression of house-keeping genes. Panepoxydone strongly inhibited hTNF-alpha, IL-8 and NF-kappaB promoter activity in LPS/TPA stimulated MonoMac6 cells with IC(50) values of 0.5-1 microg/ml by blocking the phosphorylation of IkappaB and subsequent binding of the activated NF-kappaB transcription factor to the DNA. From our data, panepoxydone may serve as lead structure for the development of transcription-based inhibitors of pro-inflammatory gene expression.
真菌次级代谢产物泛环氧薁最近被描述为一种核因子-κB（NF-κB）激活抑制剂，NF-κB是炎症和免疫反应的关键调节因子。这些发现提示泛环氧薁可能作为抗炎剂发挥作用。在本研究中，我们首次研究了泛环氧薁对脂多糖（LPS）和佛波酯12-O-十四酰佛波醇-13-乙酸酯（TPA）刺激的单核细胞系MonoMac6中炎症基因表达的影响。对110个已知在炎症过程中受到强烈调控的人类基因进行DNA微阵列分析，并结合逆转录定量实时聚合酶链反应（RT-qPCR），结果显示低微摩尔浓度（12 - 24 μM）的泛环氧薁强烈抑制33种NF-κB依赖性促炎基因的表达，如趋化因子CCL3、CCL4、CCL8；CXCL8、CXCL10、CXCL20，细胞因子IL-1、IL-6、TNF-α，促炎酶如COX-2，以及REL/NF-κB/IκB家族的成员，而对管家基因的表达无显著影响。泛环氧薁通过阻断IκB的磷酸化以及随后活化的NF-κB转录因子与DNA的结合，强烈抑制LPS/TPA刺激的MonoMac6细胞中hTNF-α、IL-8和NF-κB启动子活性，IC50值为0.5 - 1 μg/ml。根据我们的数据，泛环氧薁可能作为开发基于转录的促炎基因表达抑制剂的先导结构。
