Transcriptional regulation of the human μ-opioid receptor gene by interleukin-6
Transcriptional regulation of the human μ-opioid receptor gene by interleukin-6
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DOI：
10.1124/mol.104.003806
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发表时间：
2004-12-01
影响因子：
3.6
通讯作者：
Höllt, V
中科院分区：
文献类型：
--
作者：
Börner, C;Kraus, J;Höllt, V
炎症性疼痛是由一些生理过程抵消的。例如，存在于感觉神经元外周末梢的阿片受体被内源性阿片激活，这些阿片从免疫细胞迁移到炎症组织中释放出来。早期的数据表明，白细胞介素-6有助于这种炎症诱导的镇痛。在本报告中，我们证明了白细胞介素-6在人神经母细胞瘤细胞系SH SY5Y中强烈诱导mu-阿片受体mRNA，而delta-阿片受体mRNA水平不受影响。在这些细胞中mRNA的增加伴随着mu-阿片受体特异性结合的增加。利用转录因子诱骗寡核苷酸，直接证明完整细胞中mu-阿片受体mRNA的上调依赖于转录因子信号转导和转录激活因子1 （STAT1）和STAT3，而其他转录因子，如激活蛋白1、核因子(NF)-kappaB或NF-白细胞介素-6不参与。瞬态转染实验、电泳迁移率转移实验和转录因子诱饵寡核苷酸显示，STAT1和STAT3结合在人类μ -阿片受体基因启动子上的核苷酸- 1583位点上。对5'-TTCATGGAA-3' STAT1/3元件（回文下划线）进行突变分析，以确定STAT1和STAT3结合所需的核苷酸残基。这表明，只有回文半边和两个相邻的中心核苷酸是必需的。回文外核苷酸的突变和中心核苷酸的突变都不影响STAT1/3的结合。
Inflammatory pain is counteracted by a number of physiological processes. For example, opioid receptors, which are present on peripheral terminals of sensory neurons, are activated by endogenous opioids, which are released from immune cells migrating to the inflamed tissue. Earlier data demonstrated that interleukin-6 contributes to such inflammation-induced analgesia. In this report, we demonstrated that interleukin-6 strongly induces mu-opioid receptor mRNA in the human neuroblastoma cell line SH SY5Y, whereas delta-opioid receptor mRNA levels are not influenced. The mRNA increase in these cells is followed by an increase in mu-opioid receptor-specific binding. Using transcription factor decoy oligonucleotides, direct evidence was provided that the up-regulation of mu-opioid receptor mRNA in intact cells is dependent on the transcription factors signal transducers and activators of transcription 1 (STAT1) and STAT3, whereas other transcription factors, such as activator protein-1, nuclear factor (NF)-kappaB, or NF-interleukin-6 are not involved. STAT1 and STAT3 bound to a site located at nucleotide - 1583 on the promoter of the human mu-opioid receptor gene, as shown by transient transfection experiments, electrophoretic mobility shift assays, and transcription factor decoy oligonucleotides. A mutation analysis of the 5'-TTCATGGAA-3' STAT1/3 element ( palindrome underlined) was performed to determine nucleotide residues that are necessary for the binding of STAT1 and STAT3. It suggested that only the palindromic half sides and the two adjacent central nucleotides are required. Neither mutation of the nucleotides outside the palindrome nor mutation of the central nucleotide affected STAT1/3 binding.