Zhang Na, Yang Xin, Xu Rong, Wang Zhen, Song Dan-Qing, Li Dian-Dong, Deng Hong-Bin
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Yao Xue Xue Bao. 2013 Jul;48(7):1113-8.
LPS stimulation of macrophages production of IFN-beta plays a key role in innate immunity defending the microbial invasion. In this study, the effect of S632A3 promoting LPS-induced IFN-beta production and the underlying mechanism were investigated, mRNA level was measured by real-time PCR, cytokine production was determined by ELISA, GSK-3beta activity was investigated by kinase assay, protein phosphorylation and expression were evaluated by Western blotting. The results revealed that S632A3 significantly augmented IFN-beta production by LPS-stimulated macrophages. S632A3 inhibition of the activation of GSK-3beta, reduced the threonine 239 phosphorylation of transcription factor c-Jun but increased the total level of c-Jun in LPS-stimulated macrophages. Moreover, small interfering RNA-mediated knockdown of c-Jun level abrogated the ability of S632A3 to augment IFN-beta. The study thus demonstrates S632A3 being a new anti-inflammation lead compound and provides a molecular mechanism by which S632A3 promoted LPS-induced IFN-beta production in macrophages through inhibiting the activation of GSK-3beta.
脂多糖刺激巨噬细胞产生干扰素-β在抵御微生物入侵的天然免疫中起关键作用。在本研究中，探讨了S632A3促进脂多糖诱导的干扰素-β产生的作用及潜在机制，通过实时聚合酶链反应检测信使核糖核酸水平，采用酶联免疫吸附测定法测定细胞因子产生，通过激酶测定法研究糖原合成酶激酶-3β活性，利用蛋白质印迹法评估蛋白质磷酸化和表达情况。结果显示，S632A3显著增强了脂多糖刺激的巨噬细胞产生干扰素-β的能力。S632A3抑制糖原合成酶激酶-3β的激活，降低了脂多糖刺激的巨噬细胞中转录因子c-Jun苏氨酸239位点的磷酸化水平，但增加了c-Jun的总水平。此外，小干扰核糖核酸介导的c-Jun水平敲低消除了S632A3增强干扰素-β的能力。因此，该研究证明S632A3是一种新型抗炎先导化合物，并提供了一种分子机制，即S632A3通过抑制糖原合成酶激酶-3β的激活促进脂多糖诱导的巨噬细胞产生干扰素-β。
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