c-Jun NH2-terminal kinase inhibitor anthra(1,9-cd)pyrazol-6(2H)-one reduces inducible nitric-oxide synthase expression by destabilizing mRNA in activated macrophages
c-Jun NH2-terminal kinase inhibitor anthra(1,9-cd)pyrazol-6(2H)-one reduces inducible nitric-oxide synthase expression by destabilizing mRNA in activated macrophages
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DOI：
10.1124/mol.64.2.308
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发表时间：
2003-08-01
影响因子：
3.6
通讯作者：
Moilanen, E
中科院分区：
文献类型：
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作者：
Lahti, A;Jalonen, U;Moilanen, E
在这项研究中，我们调查的c-Jun氨基末端激酶（JNK），有丝分裂原活化蛋白激酶（MAPK）家族的成员，在脂多糖（LPS）刺激诱导型一氧化氮合酶（iNOS）的表达和一氧化氮（NO）的生产在J774小鼠巨噬细胞的作用。蒽（1，9-cd）吡唑-6（2 H）-酮（SP 600125）是JNK的药理学抑制剂，可抑制c-Jun的磷酸化，IC 50为5 - 10 μ M。在相同浓度下，SP 600125抑制LPS诱导的iNOS蛋白表达和NO产生。SP 600125对核因子kappaB的活化没有影响，而kappaB是iNOS表达的重要转录因子。如果在LPS后4 h测量，则SP 600125对iNOSmRNA水平没有显著影响。相比之下，当在LPS后8小时测量时，SP 600125使iNOSmRNA水平降低>90%。这些数据表明，SP 600125降低了iNOS mRNA的稳定性，这在使用放线菌素D的mRNA降解试验中得到了证实，其中SP 600125将iNOS mRNA半衰期从5小时降低至2小时。这些结果表明，JNK通路参与了LPS诱导的iNOS表达和NO产生的上调，其机制与稳定iNOS mRNA有关。
In this study, we investigated the role of c-Jun NH2-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family, in lipopolysaccharide (LPS)-stimulated inducible nitric-oxide synthase (iNOS) expression and nitric oxide (NO) production in J774 murine macrophages. Anthra(1,9-cd)pyrazol-6(2H)-one (SP600125), a pharmacological inhibitor of JNK, inhibited phosphorylation of c-Jun with an IC50 of 5 to 10 muM. At the same concentrations, SP600125 inhibited LPS-induced iNOS protein expression and NO production. SP600125 had no effect on the activation of nuclear factor kappaB, which is an important transcription factor for iNOS expression. SP600125 had no significant effect on iNOS mRNA levels if measured 4 h after LPS. In contrast, SP600125 reduced iNOS mRNA levels >90% when measured 8 h after LPS. These data suggest that SP600125 reduced iNOS mRNA stability, and this was confirmed in the mRNA degradation assay using actinomycin D, in which SP600125 reduced the iNOS mRNA half-life from 5 to 2 h. These results show that the JNK pathway is involved in the up-regulation of LPS-induced iNOS expression and NO production by a mechanism related to the stabilization of iNOS mRNA.