Morphine Negatively Regulates Interferon-γ Promoter Activity in Activated Murine T Cells through Two Distinct Cyclic AMP-dependent Pathways*
Morphine Negatively Regulates Interferon-γ Promoter Activity in Activated Murine T Cells through Two Distinct Cyclic AMP-dependent Pathways*
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DOI：
10.1074/jbc.m301224200
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发表时间：
2003-09
影响因子：
4.8
通讯作者：
Jinghua Wang;R. Barke;R. Charboneau;H. Loh;Sabita Roy
中科院分区：
文献类型：
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作者：
Jinghua Wang;R. Barke;R. Charboneau;H. Loh;Sabita Roy
为了探讨吗啡促进HIV感染的机制，我们研究了吗啡对野生型和μ-阿片受体敲除小鼠活化T细胞中γ-干扰素（IFN-γ）启动子的调节作用。我们的研究结果表明，吗啡抑制抗CD 3/CD 28刺激的IFN-γ启动子活性，并呈剂量依赖性。慢性吗啡处理的T细胞增加细胞内cAMP。为探讨cAMP在吗啡调节功能中的作用，研究了二丁酰环腺苷酸（cAMP）和毛喉素（forskolin）的作用。二丁酰环腺苷酸和毛喉素处理抑制IFN-γ启动子活性。用百日咳毒素治疗，而不是用蛋白激酶A抑制剂，拮抗吗啡的抑制作用。吗啡抑制ERK 1/2和p38 MAPK的磷酸化;此外，在ERK 1/2或p38 MAPK抑制剂（PD 98059或SB 203580）存在下的吗啡处理导致IFN-γ启动子活性的叠加抑制。吗啡对转录因子激活蛋白-1、NF-κB和活化T细胞核因子（NFAT）的表达有负调节作用。NF-κB p65的过表达可解除吗啡对IFN-γ启动子活性的抑制作用。但只有当NFATc 1与c-fos共过表达时，吗啡对IFN-γ启动子的抑制作用才被抵消。在μ-阿片受体敲除小鼠获得的T细胞中没有观察到吗啡的抑制作用，这表明吗啡对IFN-γ启动子活性的调节是通过μ-阿片受体介导的。总之，我们的数据表明，吗啡调节IFN-γ启动子活性是通过两个不同的cAMP依赖性途径介导的，NF-κB信号通路和ERK 1/2，p38 MAPK，AP-1/NFAT通路。
To explore the mechanism by which morphine promotes the incidence of HIV infection, we evaluated the regulatory role of morphine on the interferon-γ (IFN-γ) promoter in activated T cells from wild type and μ-opioid receptor knockout mice. Our results show that morphine inhibited anti-CD3/CD28-stimulated IFN-γ promoter activity in a dose-dependent manner. Chronic morphine treatment of T cells increased intracellular cAMP. To evaluate the role of cAMP in morphine's modulatory function, the effects of dibutyryl cyclic AMP and forskolin were investigated. Both dibutyryl cyclic AMP and forskolin treatment inhibited IFN-γ promoter activity. Treatment with pertussis toxin, but not with a protein kinase A inhibitor, antagonized morphine's inhibitory effects. Morphine inhibited phosphorylation of ERK1/2 and p38 MAPK; in addition, morphine treatment in the presence of either ERK1/2 or p38 MAPK inhibitor (PD98059 or SB203580) resulted in an additive inhibition of IFN-γ promoter activity. The transcription factor activator protein-1, NF-κB, and nuclear factor of activated T cells (NFAT) were negatively regulated by morphine. Overexpression of NF-κB p65 rescued the inhibitory effect of morphine on IFN-γ promoter activity. However, only when NFATc1 was co-overexpressed with c-fos was the inhibitory effect of morphine on IFN-γ promoter counteracted. The inhibitory effects of morphine were not observed in T cells obtained from μ-opioid receptor knockout mice, suggesting that morphine modulation of IFN-γ promoter activity is mediated through the μ-opioid receptor. In summary, our data indicate that morphine modulation of IFN-γ promoter activity is mediated through two distinct cAMP-dependent pathways, the NF-κB signaling pathway and the ERK1/2, p38 MAPK, AP-1/NFAT pathway.