Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, 1110 N. Stonewall Ave Suite 323, Oklahoma City, OK 73117, USA.
J Neuroimmune Pharmacol. 2011 Dec;6(4):617-25. doi: 10.1007/s11481-011-9279-2. Epub 2011 May 6.
Endogenous neuropeptide orphanin FQ/nociceptin (OFQ/N) and its receptor, nociceptin orphanin FQ peptide receptor (NOPr), play a modulatory role throughout the body including nociceptive sensitivity, motor function, spatial learning, and the immune system. NOPr is an inhibitory G protein coupled receptor (GPCR) that modulates expression and release of inflammatory mediators from immune cells and in the CNS. Inhibitory GPCRs have been shown to activate the immune and central nervous system regulator, nuclear factor kappa B (NFκB), whose family consists of several subunits. When activated, NFκB translocates to the nucleus and can modify transcription. To determine if OFQ/N modulates NFκB activity, SH-SY5Y human neuroblastoma cells were treated with OFQ/N and assessed for changes in nuclear accumulation, DNA binding, and transcriptional activation. For the first time, we show that OFQ/N increases the nuclear accumulation (1.9-2.8-fold) and the DNA binding of NFκB (2.9-fold) by 2 h as determined by immunoblotting and electromobility shift assay, respectively. OFQ/N induction of NFκB binding to DNA is protein kinase C-dependent and NOPr-specific. OFQ/N stimulated binding of both NFκB p50 and p65 subunits to their consensus binding site on DNA. OFQ/N also induces transcriptional activation of an NFκB reporter gene 2.2-fold by 2 h with an EC(50) of 6.3 nM. This activation of NFκB by OFQ/N suggests a likely mechanism for its modulation of the central nervous and immune systems.
内源性神经肽孤啡肽（OFQ/N）及其受体孤啡肽 N 端前体（NOPr）在全身发挥着调节作用，包括痛觉敏感性、运动功能、空间学习和免疫系统。NOPr 是一种抑制性 G 蛋白偶联受体（GPCR），可调节免疫细胞和中枢神经系统中炎症介质的表达和释放。已表明抑制性 GPCR 可激活免疫和中枢神经系统调节剂核因子 kappa B（NFκB），其家族由几个亚基组成。当被激活时，NFκB 易位到细胞核并可以修饰转录。为了确定 OFQ/N 是否调节 NFκB 活性，用 OFQ/N 处理 SH-SY5Y 人神经母细胞瘤细胞，并评估核积累、DNA 结合和转录激活的变化。我们首次表明，OFQ/N 在 2 小时内通过免疫印迹和电泳迁移率变动分析分别将 NFκB 的核积累（增加 1.9-2.8 倍）和 DNA 结合（增加 2.9 倍）。OFQ/N 诱导的 NFκB 与 DNA 的结合依赖蛋白激酶 C 和 NOPr 特异性。OFQ/N 刺激 NFκB p50 和 p65 亚基与它们在 DNA 上的共有结合位点的结合。OFQ/N 还在 2 小时内通过 NFκB 报告基因诱导转录激活增加 2.2 倍，EC（50）为 6.3 nM。OFQ/N 对 NFκB 的这种激活表明了其对中枢神经系统和免疫系统调节的可能机制。
