Inhibition of peroxisome-proliferator-activated receptor (PPAR)α by MK886
Inhibition of peroxisome-proliferator-activated receptor (PPAR)α by MK886
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DOI：
10.1042/0264-6021:3560899
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发表时间：
2001-06-15
影响因子：
4.1
通讯作者：
Heuvel, JPV
中科院分区：
文献类型：
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作者：
Kehrer, JP;Biswal, SS;Heuvel, JPV
尽管MK 886最初被鉴定为5-脂氧合酶活化蛋白（FLAP）的抑制剂，但最近的数据表明，这种活性并不构成其诱导细胞凋亡的能力的基础[Datta，Biswal和Kehrer（1999）Biochem，J.340，371-375]。由于FLAP是脂肪酸结合蛋白，因此可以想象MK 886可能影响其他此类蛋白。被脂肪酸及其代谢物激活的核受体家族。过氧化物酶体增殖物激活受体（PPARs）。已经涉及细胞凋亡，并且可能代表MK 886的靶点。使用报告分析系统评估MK 886抑制PPAR-alpha、-beta和-gamma活性的能力（过氧化物酶体增殖物反应元件-荧光素酶），在猴肾成纤维细胞CV-1细胞、小鼠角质形成细胞308细胞和人肺腺癌A549细胞中使用瞬时转染系统，10-20 μ M MK 886抑制了约80%的PPAR α的Wy 14，643活化。在CV-1细胞中，用稳定的转染报告系统观察到MK 886对PPAR α的类似抑制。仅观察到对PPAR β和PPAR γ的最小抑制作用。MK 886通过非竞争性机制抑制PPAR α，如其对花生四烯酸与PPARs蛋白结合的影响以及使用COS-1细胞中瞬时转染报告基因试验的剂量反应研究所示。一项评估PPAR配体-受体相互作用的试验表明，MK 886阻止了活性复合物形成所需的构象变化。在小鼠原代角质形成细胞培养物中，MK 886降低了角蛋白-1（一种由PPAR α反应基因编码的蛋白质）的表达，这表明PPAR抑制对正常细胞具有功能影响。虽然Jurkat细胞表达所有的过氧化物酶体增殖物激活受体亚型，各种过氧化物酶体增殖物激活受体α和γ激动剂不能防止MK 886诱导的细胞凋亡。这与MK 886作为PPARa的非竞争性抑制剂起作用一致，但也可能表明PPARa不直接参与MK 886诱导的细胞凋亡。虽然已经鉴定了许多PPAR激活剂，但结果表明MK 886可以抑制PPAR α，使其成为第一个被鉴定具有这种作用的化合物。
Although MK886 was originally identified as an inhibitor of 5-lipoxygenase activating protein (FLAP), recent data demonstrate that this activity does not underlie its ability to induce apoptosis [Datta, Biswal and Kehrer (1999) Biochem, J. 340, 371-375], Since FLAP is a fatty-acid binding protein, it is conceivable that MK886 may affect other such proteins. A family of nuclear receptors that are activated by fatty acids and their metabolites. the peroxisome-proliferator-activated receptors (PPARs). have been implicated in apoptosis and may represent a target for MK886, The ability of MK886 to inhibit PPAR-alpha, -beta and -gamma activity was assessed using reporter assay systems (peroxisomeproliferator response element-luciferase), Using a transient transfection system in monkey kidney fibroblast CV-1 cells, mouse keratinocyte 308 cells and human lung adenocarcinoma A549 cells, 10-20 muM MK886 inhibited Wy14,643 activation of PPAR alpha by approximately 80%, Similar inhibition of PPAR alpha by MK886 was observed with a stable transfection reporter system in CV-1 cells. Only minimal inhibitory effects were seen on PPAR beta and PPAR gamma. MK886 inhibited PPAR alpha by a noncompetitive mechanism as shown by its effects on the binding of arachidonic acid to PPARs protein, and a dose-response study using a transient transfection reporter assay in COS-1 cells. An assay assessing PPAR ligand-receptor interactions showed that MK886 prevents the conformational change necessary for active-complex formation. The expression of keratin-1, a protein encoded by a PPAR alpha -responsive gene, was reduced by MK886 in a culture of mouse primary keratinocytes, suggesting that PPAR inhibition has functional consequences in normal cells. Although Jurkat cells express all PPAR isoforms, various PPAR alpha and PPAR gamma agonists were unable to prevent MK886-induced apoptosis. This is consistent with MK886 functioning as a non-competitive inhibitor of PPARa, but may also indicate that PPAR alpha is not directly involved in MK886-induced apoptosis. Although numerous PPAR activators have been identified, the results show that MK886 can inhibit PPAR alpha, making it the first compound identified to have such an effect.