Interleukin (IL)-10 inhibits IL-6 production in microglia by preventing activation of NF-κB
Interleukin (IL)-10 inhibits IL-6 production in microglia by preventing activation of NF-κB
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DOI：
10.1016/s0169-328x(00)00042-5
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发表时间：
2000-04-14
期刊：
影响因子：
--
通讯作者：
Johnson, RW
中科院分区：
文献类型：
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作者：
Heyen, JRR;Ye, SM;Johnson, RW
本研究的目的是确定白细胞介素（IL）-10是否通过抑制核因子-κ B（NF-κ B）的活化来抑制脂多糖（LPS）诱导的小胶质细胞中IL-6的产生。N13小神经胶质细胞（鼠小神经胶质细胞系）和来自新生小鼠的原代小神经胶质细胞在存在或不存在LPS和增加量的鼠IL-10的情况下培养24小时。正如所预测的，LPS处理增加了N13和原代小胶质细胞培养物中上清液IL-6的浓度。然而，在两种培养系统中，用IL-10预处理以剂量依赖性方式降低LPS诱导的IL-6分泌。同样，核糖核酸酶保护试验表明，LPS增加稳态IL-6 mRNA水平，但与IL-10预处理阻断LPS诱导的IL-6 mRNA的增加。由于NF-κ B是响应炎症刺激而负责IL-6转录的主要转录因子，因此假设IL-10通过阻止NF-κ B的核转位来抑制IL-6产生。与此一致，LPS增加NF-κ B B的核转位，如通过凝胶迁移率变动分析所评估的。超移位实验和免疫细胞化学染色显示，在LPS刺激下，NF-κ B B的p50和p65亚基均从细胞质移位到细胞核。然而，用IL-10预处理抑制LPS诱导的NF-κ B活化。此外，用甲苯磺酰基-Phe-氯-甲基酮（一种防止NF-κ B-I κ B复合物降解的丝氨酸蛋白酶抑制剂）抑制NF-κ B活性完全阻断了LPS诱导的IL-6产生。这些数据表明，IL-10通过降低NF-κ B的活性来抑制小胶质细胞中IL-6的产生，因此，扩展了对中枢神经系统中抗炎和炎性细胞因子之间的复杂关系知之甚少的内容。(C)2000 Elsevier Science B. V.保留所有权利。
The purpose of this study was to determine if interleukin (IL)-10 inhibits lipopolysaccharide (LPS)-induced IL-6 production in microglia by inhibiting activation of nuclear factor-kappa B (NF-kappa B). N13 microglia (a murine microglial cell line) and primary microglia from neonatal mice were cultured in the presence or absence of LPS and increasing amounts of murine IL-10 for 24 h. As predicted, LPS treatment increased supernatant IL-6 concentration in both N13 and primary microglia cultures. Pretreatment with IL-10, however, decreased LPS-induced IL-6 secretion in a dose-dependent manner in both culture systems. Likewise, ribonuclease protection assays showed that LPS increased steady-state IL-6 mRNA levels, but that pretreatment with IL-10 blocked the LPS-induced increase in IL-6 mRNA. Because NF-kappa B is the predominant transcription factor responsible for IL-6 transcription in response to inflammatory stimuli, it was hypothesized that IL-10 inhibited IL-6 production by preventing nuclear translocation of NF-kappa B. Consistent with this idea, LPS increased nuclear translocation of NF-kappa B as assessed by gel mobility shift assay. Supershift assays and immunocytochemical staining showed that both the p50 and p65 subunits of NF-kappa B translocated from the cytoplasm to the nucleus upon LPS stimulation. Pretreatment with IL-10, however, inhibited LPS-induced activation of NF-kappa B. Furthermore, inhibition of NF-kappa B activity with tosyl-Phe-chloro-methlyketone (a serine protease inhibitor that prevents degradation of the NF-kappa B-I kappa B complex), completely blocked LPS-induced IL-6 production. These data suggest that IL-10 inhibited IL-6 production in microglia by decreasing the activity of NF-kappa B and, therefore, extend what little is known of the intricate relationship between anti-inflammatory and inflammatory cytokines in the central nervous system. (C) 2000 Elsevier Science B.V. All rights reserved.