Abstract
A number of drugs were assessed for their ability to inhibit stimulus-evoked prostanoid synthesis in cultured glia. These drugs included non-selective cyclooxygenase (COX) inhibitors and those considered to be selective for the inducible isoform of this enzyme (COX-2). Experiments were carried out on normal cultures and those which had been maintained in serum-free growth medium for four days then re-exposed to serum for a further seven days. All of the drugs tested elicited concentration-dependent inhibitions of arachidonic acid (AA)-stimulated thromboxane B2 (TXB2) accumulation in normal cultures with the following rank order of potency: indomethacin > piroxicam > nimesulide = NS398 > ibuprofen ≫ aspirin > paracetamol. In cultures which had been deprived of serum for four days, basal and AA-stimulated TXB2 production was considerably reduced, as was the amount of COX immunoreactivity determined by Western blotting. Basal and AA-stimulated TXB2 production together with COX immunoreactivity were restored to control levels by the re-addition of serum to serum-deprived cultures for 7 days. In these cultures, the rank order of potency was: indomethacin > piroxicam ≫ ibuprofen > nimesulide = NS398 ≫ aspirin > paracetamol; however, there were marked charges in the apparent IC50 values for particular drugs. Indomethacin, piroxicam and aspirin were very similar to control, but the potencies of ibuprofen (3-fold), NS398 (30-fold) and nimesulide (40-fold) were found to be decreased when compared to control. Paracetamol, on the other hand, was found to be almost 3-fold more potent under these conditions. Glia appear to express a COX with a novel sensitivity to particular inhibitors following serum deprivation and re-addition.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Breder, C. D., DeWitt, D. and Kraig, R. P. (1995). Characterization of inducible cyclooxygenase in rat brain, J. Comp. Neurol. 355, 296–315.
Buccellati, C., Folco, G. C., Sala, A., et al. (1998). Inhibition of prostanoid synthesis protects against neuronal damage induced by focal ischemia in rat brain, Neurosci. Lett. 257, 123–126.
Chandrasekharan, N. V., Dai, H., Roos, K. L. T., et al. (2002). COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and other analgesic/antipyretic drugs: cloning structure and function, Proc. Natl. Acad. Sci. USA 99, 13926–13931.
Chen, S. T., Hsu, C. Y., Hogan, E. L., et al. (1986). Thromboxane, prostacyclin and leukotrienes in cerebral ischemia, Neurology 36, 466–470.
Degi, R., Bari, F., Beasley, T. C., et al. (1998). Regional distribution of prostaglandin H synthase-2 and neuronal nitric oxide synthase in piglet brain, Pediat. Res. 43, 683–689.
Eddleston, M. and Mucke, L. (1993). Molecular profile of reactive astrocytes — implications for their role in neurologic disease, Neuroscience 54, 15–36.
Gebicke-Haerter, P. J., Bauer, J., Schobert, A., et al. (1989). Lipopolysaccharide-free conditions in primary astrocyte cultures allow growth and isolation of microglial cells, J. Neurosci. 9, 183–194.
Govoni, S., Masoero, E., Favalli, L., et al. (2001). The cyclooxygenase-2 inhibitor SC58236 is neuroprotective in an in vivo model of focal ischaemia in the rat, Neurosci. Lett. 303, 91–94.
Hirst, W. D., Young, K. A., Newton, R., et al. (1999). Expression of COX-2 by normal and reactive astrocytes in the adult rat central nervous system, Mol. Cell. Neurosci. 13, 57–68.
Huttemeier, P. C., Kamiyama, Y., Su, M., et al. (1993). Microdialysis measurements of PGD2, TXB2 and 6-keto-PGF1α in rat CA1 hippocampus during transient cerebral ischaemia, Prostaglandins 45, 177–187.
Iadecola, C., Forster, C., Nogawa, S., et al. (1995). Cyclooxygenase-2 immunoreactivity in the human brain following cerebral ischaemia, Acta Neuropathol. 98, 9–14.
Inoue, H., Yokoyama, C., Hara, S., et al. (1995). Transcriptional regulation of human prostaglandinendoperoxide synthase-2 gene by lipopolysaccharide and phorbol ester in vascular endothelial cells, J. Biol. Chem. 270, 24965–24971.
Jeremy, J., Murphy, S., Morrow, C., et al. (1987). Phorbol ester stimulation of prostanoid synthesis by cultured astrocytes, Brain Res. 419, 364–368.
Kawai, S., Nishida, S., Kato, M., et al. (1998). Comparison of cyclooxygenase-1 and-2 inhibitory activities of various non-steroidal anti-inflammatory drugs using human platelets and synovial cells, Eur. J. Pharmacol. 347, 87–94.
Kis, B., Snipes, J. A., Isse, T., et al. (2003). Putative cyclooxygenase-3 expression in rat brain cells, J. Cereb. Blood Flow Metab. 23, 1287–1292.
Luo, J., Lang, J. A. and Miller, M. W. (1998). Transforming growth factor β1 regulates the expression of cyclooxygenase in cultured cortical astrocytes and neurons, J. Neurochem. 71, 526–534.
Maslinska, D., Wozniak, R., Kaliszek, A., et al. (1997). Expression of cyclooxygenase-2 in astrocytes of human brain after global ischemia, Folia Neuropathol. 37, 75–79.
Miettinen, S., Fusco, F. R., Yrjanheikki, J., et al. (1997). Spreading depression and focal brain ischaemia induce cyclooxygenase-2 in cortical neurons through N-methyl-d-aspartic acid receptors and phospholipase A2, Proc. Natl. Acad. Sci. USA 94, 6500–6505.
Mitchell, J. A., Larkin, S. and Williams, T. J. (1995). Cyclooxygenase-2: Regulation and relevance in inflammation, Biochem. Pharmacol. 50, 1535–1542.
Murphy, S., Pearce, B., Jeremy, J. Y., et al. (1988). Astrocytes as eicosanoid-producing cells, Glia 1, 241–245.
Nakayama, M., Uchimura, K., Zhu, R. L., et al. (1998). Cyclooxygenase-2 inhibition prevents delayed death of CA1 hippocampal neurons following global ischaemia, Proc. Natl. Acad. Sci. USA 95, 10954–10959.
Nogawa, S., Zhang, F., Ross, M. E., et al. (1997). Cyclooxygenase-2 gene expression in neurons contributes to ischaemic brain damage, J. Neurosci. 17, 2746–2755.
O’Banion, M. K., Miller, J. C., Chang, J. W., et al. (1996). Interleukin-1β induces prostaglandin G/H synthase-2 (cyclooxygenase-2) in primary murine astrocyte cultures, J. Neurochem. 66, 2532–2540.
Rich, G., Yoder, E. J. and Moore, S. A. (1998). Regulation of prostaglandin H synthase-2 expression in cerebromicrovascular smooth muscle by serum and epidermal growth factor, J. Cell Physiol. 176, 495–505.
Sairanen, T., Ristimaki, A., Karjalainen-Lindsberg, M. L., et al. (1998). Cyclooxygenase-2 is induced globally in infarcted human brain, Ann. Neurol. 43, 738–747.
Sandhya, T. L., Ong, W. Y., Horrocks, L. A., et al. (1998). A light and electron microscopic study of cytoplasmic phospholipase A2 and cyclooxygenase-2 in the hippocampus after kainate lesions, Brain Res. 788, 223–231.
Shaftel, S. S., Olschowka, J. A., Hurley, S. D., et al. (2003). COX-3: a splice variant of cyclooxygenase-1 in mouse neural tissue and cells, Mol. Brain Res. 119, 213–215.
Simmons, D. L., Botting, R. M., Robertson, P. M., et al. (1999). Induction of an acetaminophensensitive cyclooxygenase with reduced sensitivity to non-steroid anti-inflammatory drugs, Proc. Natl. Acad. Sci. USA 96, 3275–3280.
Stoll, G., Jander, S. and Schroeter, M. (1998). Inflammation and glial responses in ischaemic brain lesions, Prog. Neurobiol. 56, 149–171.
Tomimoto, H., Akiguchi, I., Wakita, H., et al. (2000). Cyclooxygenase-2 is induced in microglia during chronic cerebral ischaemia, Acta Neuropathol. 99, 26–30.
Vane, J. R., Bakhle, Y. S. and Botting, R. M. (1998). Cyclooxygenases 1 and 2, Annu. Rev. Pharmacol. Toxicol. 38, 97–120.
Warner, T. D. and Mitchell, J. A. (2002). Cyclooxygenase-3 (COX-3): Filling in the gaps towards a COX continuum?, Proc. Natl. Acad. Sci. USA 99, 13371–13373.
Warner, T. D., Giuliano, F., Vojnovic, I., et al. (1999). Nonsteroid drug selectivities for cyclo-oxygenase-1 rather than cyclo-oxygenase-2 are associated with human gastrointestinal toxicity: a full in vitro analysis, Proc. Natl. Acad. Sci. USA 96, 7563–7568.
Xie, W. and Herschman, H. R. (1996). Transcriptional regulation of prostaglandin synthase 2 gene expression by platelet-derived growth factor and serum, J. Biol. Chem. 271, 31742–31748.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Phillips, J., Pearce, B. Serum deprivation and re-addition: effects on cyclooxygenase inhibitor sensitivity in cultured glia. Inflammopharmacol 13, 431–439 (2005). https://doi.org/10.1163/156856005774649368
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1163/156856005774649368