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Ischemic Preconditioning Mediates Cyclooxygenase-2 Expression Via Nuclear Factor-Kappa B Activation in Mixed Cortical Neuronal Cultures

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Abstract

Nuclear factor-kappaB (NF-κB) activation occurs following ischemic preconditioning (IPC) in brain. However, the upstream signaling messengers and down-stream targets of NF-κB required for induction of IPC remain undefined. In a previous study, we demonstrated that epsilon protein kinase c (εPKC) was a key mediator of IPC in brain. Activation of εPKC induced cyclooygenase-2 (COX-2) expression and conferred ischemic tolerance in the neuronal and hippocampal slice models. Here, we hypothesized that IPC-mediated COX-2 expression was mediated by NF-κB. We tested this hypothesis in mixed cortical neuron/astrocyte cell cultures. To simulate IPC or ischemia, cell cultures were exposed to 1 or 4 h of oxygen–glucose deprivation, respectively. Our results demonstrated translocation of p65 and p50 subunits of NF-κB into nucleus following IPC or εPKC activation. NF-κB inhibition with pyrrolidine dithiocarbamate (10 μM) abolished IPC or εPKC activator-mediated neuroprotection indicating that NF-κB activation was involved in ischemic tolerance. In parallel studies, inhibition of either εPKC or the extracellular signal-regulated kinase (ERK 1/2) pathway reduced IPC-induced NF-κB activation. Finally, inhibition of NF-κB blocked IPC-induced COX-2 expression. In conclusion, we demonstrated that IPC-signaling cascade comprises εPKC activation → ERK1/2 activation → NF-κB translocation to nucleus → COX-2 expression resulting in neuroprotection in mixed neuronal culture.

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Acknowledgments

This study was supported by PHS grants NS34773, NS054147, NS045676 (MAPP), AHA-National center # 0730089 N, the James and Esther King Biomedical Research Program, Florida Department of Health 07KN-10 (APR), and AHA Florida & Puerto Rico Affiliate grant 0525331B (MAPP/EJ).

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Authors and Affiliations

  1. Cerebral Vascular Disease Research Center, Department of Neurology and Neuroscience Program (D4-5), University of Miami Miller School of Medicine, P.O. Box 016960, Miami, FL, 33101, USA

    Eun Joo Kim, Ami P. Raval, Nina Hirsch & Miguel A. Perez-Pinzon

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  1. Eun Joo Kim
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  2. Ami P. Raval
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  3. Nina Hirsch
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  4. Miguel A. Perez-Pinzon
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Correspondence to Miguel A. Perez-Pinzon.

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Kim, E.J., Raval, A.P., Hirsch, N. et al. Ischemic Preconditioning Mediates Cyclooxygenase-2 Expression Via Nuclear Factor-Kappa B Activation in Mixed Cortical Neuronal Cultures. Transl. Stroke Res. 1, 40–47 (2010). https://doi.org/10.1007/s12975-009-0006-8

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