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Orphanin FQ/Nociceptin Activates Nuclear Factor Kappa B

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Abstract

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 EC50 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.

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Acknowledgments

The research results discussed in this publication were made possible, in part, by the OHRS award for project number HR08-152 from the Oklahoma Center for the Advancement of Science and Technology, grant number R01DA017380 from the National Institute on Drug Abuse, and the Department of the Army (W81XWH-09-1-0443; The U.S. Army Medical Research Acquisition Activity, 820 Chandler Street, Fort Detrick MD 21702-5014 is the awarding and administering acquisition office). The information and interpretations included in this manuscript do not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. The authors would like to thank Andrew Mansour and Ryan Webb for excellent technical and statistical assistance, respectively.

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Correspondence to Kelly M. Standifer.

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Guarantors: Courtney L. Donica and Kelly M. Standifer.

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Donica, C.L., Ramirez, V.I., Awwad, H.O. et al. Orphanin FQ/Nociceptin Activates Nuclear Factor Kappa B. J Neuroimmune Pharmacol 6, 617–625 (2011). https://doi.org/10.1007/s11481-011-9279-2

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