Ascorbic acid enhances the inhibitory effect of aspirin on neuronal cyclooxygenase-2-mediated prostaglandin E2 production.

Candelario-Jalil, Eduardo and Akundi, Ravi S. and Bhatia, Harsharan S. and Lieb, Klaus and Appel, Kurt and Munoz, Eduardo and Hull, Michael and Fiebich, Bernd L. (2006) Ascorbic acid enhances the inhibitory effect of aspirin on neuronal cyclooxygenase-2-mediated prostaglandin E2 production. [Journal (Paginated)]

Full text available as:



Inhibition of neuronal cyclooxygenase-2 (COX-2) and hence prostaglandin E2 (PGE2) synthesis by non-steroidal anti-inflammatory drugs has been suggested to protect neuronal cells in a variety of pathophysiological situations including Alzheimer's disease and ischemic stroke. Ascorbic acid (vitamin C) has also been shown to protect cerebral tissue in a variety of experimental conditions, which has been attributed to its antioxidant capacity. In the present study, we show that ascorbic acid dose-dependently inhibited interleukin-1beta (IL-1beta)-mediated PGE2 synthesis in the human neuronal cell line, SK-N-SH. Furthermore, in combination with aspirin, ascorbic acid augmented the inhibitory effect of aspirin on PGE2 synthesis. However, ascorbic acid had no synergistic effect along with other COX inhibitors (SC-58125 and indomethacin). The inhibition of IL-1beta-mediated PGE2 synthesis by ascorbic acid was not due to the inhibition of the expression of COX-2 or microsomal prostaglandin E synthase (mPGES-1). Rather, ascorbic acid dose-dependently (0.1-100 microM) produced a significant reduction in IL-1beta-mediated production of 8-iso-prostaglandin F2alpha (8-iso-PGF2alpha), a reliable indicator of free radical formation, suggesting that the effects of ascorbic acid on COX-2-mediated PGE2 biosynthesis may be the result of the maintenance of the neuronal redox status since COX activity is known to be enhanced by oxidative stress. Our results provide in vitro evidence that the neuroprotective effects of ascorbic acid may depend, at least in part, on its ability to reduce neuronal COX-2 activity and PGE2 synthesis, owing to its antioxidant properties. Further, these experiments suggest that a combination of aspirin with ascorbic acid constitutes a novel approach to render COX-2 more sensitive to inhibition by aspirin, allowing an anti-inflammatory therapy with lower doses of aspirin, thereby avoiding the side effects of the usually high dose aspirin treatment.

Item Type:Journal (Paginated)
Keywords:vitamin C, cyclooxygenase-2, oxidative stress, Alzheimer’s disease, stroke, reactive oxygen species, neuroinflammation, NSAIDs, 8-isoprostanes, prostaglandin E synthase
Subjects:Neuroscience > Neurochemistry
ID Code:5641
Deposited By: Candelario-Jalil, Dr Eduardo
Deposited On:03 Aug 2007
Last Modified:11 Mar 2011 08:56

References in Article

Select the SEEK icon to attempt to find the referenced article. If it does not appear to be in cogprints you will be forwarded to the paracite service. Poorly formated references will probably not work.

Abate, A., Yang, G., Dennery, P.A., Oberle, S., Schroder, H. 2000. Synergistic inhibition of cyclooxygenase-2 expression by vitamin E and aspirin. Free Radic. Biol. Med. 29, 1135-1142.

Akundi, R. S., Candelario-Jalil, E., Hess, S., Hüll, M., Lieb, K., Gebicke-Haerter, P. J., Fiebich, B. L. 2005. Signal transduction pathways regulating cyclooxygenase-2 in lipopolysaccharide-activated primary rat microglia. Glia 51, 199-208.

Andersen, J.K. 2004. Oxidative stress in neurodegeneration: cause or consequence? Nat. Med. 10 Suppl, S18-S25.

Basu, A., Krady, J.K., Levison, S.W. 2004. Interleukin-1: a master regulator of neuroinflammation. J. Neurosci. Res. 78, 151-156.

Becker, J.C., Grosser, N., Boknik, P., Schroder, H., Domschke, W., Pohle T. 2003. Gastroprotection by vitamin C-a heme oxygenase-1-dependent mechanism?. Biochem. Biophys. Res. Commun. 312, 507-512.

Beharka, A.A., Wu, D., Serafini, M., Meydani, S.N. 2002. Mechanism of vitamin E inhibition of cyclooxygenase activity in macrophages from old mice: role of peroxynitrite. Free Radic. Biol. Med. 32, 503-511.

Boutaud, O., Aronoff, D.M., Richardson, J.H., Marnett, L.J., Oates, J.A. 2002. Determinants of the cellular specificity of acetaminophen as an inhibitor of prostaglandin H(2) synthases. Proc. Natl. Acad. Sci. U. S. A 99, 7130-7135.

Braddock, M., Quinn, A. 2004. Targeting IL-1 in inflammatory disease: new opportunities for therapeutic intervention. Nat. Rev. Drug Discov. 3, 330-339.

Brzozowski, T., Kwiecien, S., Konturek, P.C., Konturek, S.J., Mitis-Musiol, M., Duda, A., Bielanski, W., Hahn, E.G. 2001. Comparison of nitric oxide-releasing NSAID and vitamin C with classic NSAID in healing of chronic gastric ulcers; involvement of reactive oxygen species. Medicinal Science Monitoring 7, 592-599.

Candelario-Jalil, E., Gonzalez-Falcon, A., Garcia-Cabrera, M., Alvarez, D., Al Dalain, S., Martinez, G., Leon, O.S., Springer, J.E. 2003a. Assessment of the relative contribution of COX-1 and COX-2 isoforms to ischemia-induced oxidative damage and neurodegeneration following transient global cerebral ischemia. J. Neurochem. 86, 545-555.

Candelario-Jalil, E., Alvarez, D., Merino, N., Leon, O.S. 2003b. Delayed treatment with nimesulide reduces measures of oxidative stress following global ischemic brain injury in gerbils. Neurosci. Res. 47, 245-253.

Candelario-Jalil, E., Gonzalez-Falcon, A., Garcia-Cabrera, M., Leon, O.S., Fiebich, B.L. 2004. Wide therapeutic time window for nimesulide neuroprotection in a model of transient focal cerebral

ischemia in the rat. Brain Res. 1007, 98-108.

Chang, J.W., Coleman, P.D., O'Banion, M.K. 1996. Prostaglandin G/H synthase-2 (cyclooxygenase-2) mRNA expression is decreased in Alzheimer's disease. Neurobiol. Aging 17, 801-808.

Chomczynski, P., Sacchi, N. 1987. Single-step method of RNA isolation by acid guanidinium thiocyanatephenol-chloroform extraction. Ann. Biochem. 162, 156-157.

Ciuffi, M., Tarlini, L., Mugnai, S., Franchi-Micheli, S., Zilletti, L. 1996. Hemoglobin affects lipid peroxidation and prostaglandin E2 formation in rat corticocerebral tissues in vitro. Biochem. Pharmacol. 52, 97-103.

Dammann, H.G., Saleki, M., Torz, M., Schulz, H.U., Krupp, S., Schurer, M., Timm, J., Gessner, U. 2004. Effects of buffered and plain acetylsalicylic acid formulations with and without ascorbic acid on gastric mucosa in healthy subjects. Aliment. Pharmacol. Ther. 19, 367-374.

Demasi, M., Caughey, G.E., James, M.J., Cleland, L.G. 2000. Assay of cyclooxygenase-1 and 2 in human monocytes. Inflamm. Res. 49, 737-743.

DeWitt, D.L. 1999. Cox-2-selective inhibitors: the new super aspirins. Mol. Pharmacol. 55, 625-631.

DeWitt, D.L., el-Harith, E.A., Kraemer, S.A., Andrews, M.J., Yao, E.F., Armstrong, R.L., Smith, W.L. 1990. The aspirin and heme-binding sites of ovine and murine prostaglandin endoperoxide synthases. J. Biol. Chem. 265, 5192-5198.

Engelhart, M.J., Geerlings, M.I., Ruitenberg, A., van Swieten, J.C., Hofman, A., Witteman, J.C., Breteler, M.M. 2002. Dietary intake of antioxidants and risk of Alzheimer disease. JAMA 287, 3223-3229.

Fiebich, B.L., Lieb, K., Hull, M., Aicher, B., van Ryn, J., Pairet, M., Engelhardt, G. 2000a. Effects of caffeine and paracetamol alone or in combination with acetylsalicylic acid on prostaglandin E(2) synthesis in rat microglial cells. Neuropharmacology 39, 2205-2213.

Fiebich, B.L., Lieb, K., Kammerer, N., Hüll, M. 2003. Synergistic inhibitory effect of ascorbic acid and acetylsalicylic acid on prostaglandin E2 release in primary rat microglia. J. Neurochem. 86, 173-178.

Fiebich, B.L., Mueksch, B., Boehringer, M., Hull, M. 2000b. Interleukin-1beta induces cyclooxygenase-2 and prostaglandin E(2) synthesis in human neuroblastoma cells: involvement of p38 mitogen-activated protein kinase and nuclear factor-kappaB. J. Neurochem. 75, 2020-2028.

Grunewald, R.A. 1993. Ascorbic acid in the brain. Brain Res. Brain Res. Rev. 18, 123-133.

Han, S.S., Kim, K., Hahm, E.R., Lee, S.J., Surh, Y.J., Park, H.K., Kim, W.S., Jung, C.W., Lee, M.H., Park, K., Yang, J.H., Yoon, S.S., Riordan, N.H., Riordan, H.D., Kimler, B.F., Park, C.H., Lee, J.H., Park, S. 2004. L-ascorbic acid represses constitutive activation of NF-kappaB and COX-2 expression in human acute myeloid leukemia, HL-60. J. Cell Biochem. 93, 257-270.

Harris, R.E., Namboodiri, K.K., Farrar, W.B. 1996. Nonsteroidal antiinflammatory drugs and breast cancer. Epidemiology 7, 203-205.

Hemler, M.E., Lands, W.E. 1980. Evidence for a peroxide-initiated free radical mechanism of prostaglandin biosynthesis. J. Biol. Chem. 255, 6253-6261.

Ho, L., Pieroni, C., Winger, D., Purohit, D.P., Aisen, P.S., Pasinetti, G.M. 1999. Regional distribution of cyclooxygenase-2 in the hippocampal formation in Alzheimer's disease. J. Neurosci. Res. 57, 295-303.

Hull, M., Lieb, K., Fiebich, B.L. 2002. Pathways of inflammatory activation in Alzheimer's disease: potential targets for disease modifying drugs. Curr. Med. Chem. 9, 83-88.

Iadecola, C., Niwa, K., Nogawa, S., Zhao, X., Nagayama, M., Araki, E., Morham, S., Ross, M.E. 2001. Reduced susceptibility to ischemic brain injury and N-methyl-D-aspartate-mediated neurotoxicity in cyclooxygenase-2-deficient mice. Proc. Natl. Acad. Sci. USA 98, 1294-1299.

Igwe, O.J., Murray, J.N., Moolwaney, A.S. 2001. Interleukin 1-induced cyclooxygenase and nitric oxide synthase gene expression in the rat dorsal root ganglia is modulated by antioxidants. Neuroscience 105, 971-985.

in t'Velt, B., Ruitenberg, A., Hofman, A., Launer, L.J., Van Duijn, C.M., Stijnen, T., Breteler, M.M., Stricker, B.H. 2001. Nonsteroidal antiinflammatory drugs and the risk of Alzheimer's disease. N. Engl. J. Med. 345, 1515-1521.

Jakobsson, P.J., Thoren, S., Morgenstern, R., Samuelsson, B. 1999. Identification of human prostaglandin E synthase: a microsomal, glutathione-dependent, inducible enzyme, constituting a potential novel drug target. Proc. Natl. Acad. Sci. U. S. A 96, 7220-7225.

Jiang, J., Borisenko, G.G., Osipov, A., Martin, I., Chen, R., Shvedova, A.A., Sorokin, A., Tyurina, Y.Y., Potapovich, A., Tyurin, V.A., Graham, S.H., Kagan, V.E. 2004. Arachidonic acid-induced carbon centered radicals and phospholipid peroxidation in cyclo-oxygenase-2-transfected PC12 cells. J. Neurochem. 90, 1036-1049.

Jiang, Q., Elson-Schwab, I., Courtemanche, C., Ames, B.N. 2000. gamma-tocopherol and its major metabolite, in contrast to alpha-tocopherol, inhibit cyclooxygenase activity in macrophages and epithelial cells. Proc. Natl. Acad. Sci. U. S. A 97, 11494-11499.

Juni, P., Nartey, L., Reichenbach, S., Sterchi, R., Dieppe, P.A., Egger, M. 2004. Risk of cardiovascular events and rofecoxib: cumulative meta-analysis. Lancet 364, 2021-2029.

Kaufmann, W.E., Worley, P.F., Pegg, J., Bremer, M., Isakson, P. 1996. COX-2, a synaptically induced enzyme, is expressed by excitatory neurons at postsynaptic sites in rat cerebral cortex. Proc. Natl. Acad. Sci. U. S. A. 93, 2317-2321.

Kelley, K.A., Ho, L., Winger, D., Freire-Moar, J., Borelli, C.B., Aisen, P.S., Pasinetti, G.M. 1999. Potentiation of excitotoxicity in transgenic mice overexpressing neuronal cyclooxygenase-2. Am. J. Pathol. 155, 995-1004.

Kim, H.J., Kim, K.W., Yu, B.P., Chung, H.Y. 2000. The effect of age on cyclooxygenase-2 gene expression: NF-kappaB activation and IkappaBalpha degradation. Free Radic. Biol. Med. 28, 683-692.

Konturek, P.C., Kania, J., Gessner, U., Konturek, S.J., Hahn, E.G., Konturek, J.W. 2004. Effect of vitamin C releasing acetylsalicylic acid on gastric mucosal damage before and after Helicobacter pylori eradication therapy. Eur. J. Pharmacol. 506, 169-177.

Kukreja, R.C., Kontos, H.A., Hess, M.L., Ellis, E.F. 1986. PGH synthase and lipoxygenase generate superoxide in the presence of NADH or NADPH. Circ. Res. 59, 612-619.

Kulmacz, R.J., Lands, W.E. 1983. Requirements for hydroperoxide by the cyclooxygenase and peroxidase activities of prostaglandin H synthase. Prostaglandins 25, 531-540.

Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227, 680-685.

Lecomte, M., Laneuville, O., Ji, C., DeWitt, D.L., Smith, W.L. 1994. Acetylation of human prostaglandin endoperoxide synthase-2 (cyclooxygenase-2) by aspirin. J. Biol. Chem. 269, 13207-13215.

Levine, M., Conry-Cantilena, C., Wang, Y., Welch, R.W., Washko, P.W., Dhariwal, K.R., Park, J.B., Lazarev, A., Graumlich, J.F., King, J., Cantilena, L.R. 1996. Vitamin C pharmacokinetics in healthy

volunteers: evidence for a recommended dietary allowance. Proc. Natl. Acad. Sci. USA 93, 3704-3709.

Luong, C., Miller, A., Barnett, J., Chow, J., Ramesha, C., Browner, M.F. 1996. Flexibility of the NSAID binding site in the structure of human cyclooxygenase-2. Nature Struct. Biol. 3, 927-933.

Madrigal, J.L., Moro, M.A., Lizasoain, I., Lorenzo, P., Fernandez, A.P., Rodrigo, J., Bosca, L., Leza, J.C. 2003. Induction of cyclooxygenase-2 accounts for restraint stress-induced oxidative status in rat brain. Neuropsychopharmacology 28, 1579-1588.

Mancini, J.A., Blood, K., Guay, J., Gordon, R., Claveau, D., Chan, C.C., Riendeau, D. 2001. Cloning, expression, and up-regulation of inducible rat prostaglandin E synthase during lipopolysaccharide-induced pyresis and adjuvant-induced arthritis. J. Biol. Chem. 276, 4469-4475.

Mancini, J.A., Vickers, P.J., O'Neill, G.P., Boily, C., Falgueyret, J.P., Riendeau, D. 1997. Altered sensitivity of aspirin-acetylated prostaglandin G/H synthase-2 to inhibition by nonsteroidal anti-inflammatory drugs. Mol. Pharmacol. 51, 52-60.

May, J.M. 2002. Recycling of vitamin C by mammalian thioredoxin reductase. Methods Enzymol. 347, 327-332.

May, J.M., Mendiratta, S., Hill, K.E., Burk, R.F. 1997. Reduction of dehydroascorbate to ascorbate by the selenoenzyme thioredoxin reductase. J. Biol. Chem. 272, 22607-22610.

Miele, M., Fillenz, M. 1996. In vivo determination of extracellular brain ascorbate. J. Neurosci. Methods 70, 15-19.

Miettinen, S., Fusco, F.R., Yrjänheikki, J., Keinänen, R., Hirovonen, T., Roivainen, R., Närhi, M., Hökfelt, T., Koistinaho, J. 1997. Spreading depression and focal brain ischemia induce cyclooxygenase-2 in cortical neurons through N-methyl-D- aspartic acid-receptors and phospholipase A2. Proc. Natl. Acad. Sci. U. S. A. 94, 6500-6505.

Murakami, M., Nakatani, Y., Tanioka, T., Kudo, I. 2002. Prostaglandin E synthase. Prostaglandins Other Lipid Mediat. 68-69, 383-399.

Murakami, M., Naraba, H., Tanioka, T., Semmyo, N., Nakatani, Y., Kojima, F., Ikeda, T., Fueki, M., Ueno, A., Oh, S., Kudo, I. 2000. Regulation of prostaglandin E2 biosynthesis by inducible membraneassociated prostaglandin E2 synthase that acts in concert with cyclooxygenase-2. J. Biol. Chem. 275, 32783-32792.

Ohtsuki, T., Kitagawa, K., Yamagata, K., Mandai, K., Mabuchi, T., Matsushita, K., Yanagihara, T., Matsumoto, M. 1996. Induction of cyclooxygenase-2 mRNA in gerbil hippocampal neurons after

transient forebrain ischemia. Brain Res. 736, 353-356.

Oka, A., Takashima, S. 1997. Induction of cyclo-oxygenase 2 in brains of patients with Down's syndrome and dementia of Alzheimer type: specific localization in affected neurones and axons. Neuroreport. 8, 1161-1164.

Pairet, M., Engelhardt, G. 1996. Distinct isoforms (COX-1 and COX-2) of cyclooxygenase: possible physiological and therapeutic implications. Fundam. Clin. Pharmacol. 10, 1-17.

Pasinetti, G.M. 2001. Cyclooxygenase and Alzheimer's disease: implications for preventive initiatives to slow the progression of clinical dementia. Arch. Gerontol. Geriatr. 33, 13-28.

Pasinetti, G.M., Aisen, P.S. 1998. Cyclooxygenase-2 expression is increased in frontal cortex of Alzheimer's disease brain. Neuroscience 87, 319-324.

Pepicelli, O., Fedele, E., Bonanno, G., Raiteri, M., Ajmone-Cat, M.A., Greco, A., Levi, G., Minghetti, L. 2002. In vivo activation of N-methyl-D-aspartate receptors in the rat hippocampus increases

prostaglandin E(2) extracellular levels and triggers lipid peroxidation through cyclooxygenase-mediated mechanisms. J. Neurochem. 81, 1028-1034.

Pohle, T., Brzozowski, T., Becker, J.C., Van der Voort, I.R., Markmann, A., Konturek, S.J., Moniczewski, A., Domschke, W., Konturek, J.W. 2001. Role of reactive oxygen metabolites in aspirin-induced gastric damage in humans: gastroprotection by vitamin C. Aliment. Pharmacol. Ther. 15, 677-687.

Pratico, D., Barry, O.P., Lawson, J.A., Adiyaman, M., Hwang, S.W., Khanapure, S.P., Iuliano, L., Rokach, J., Fitzgerald, G.A. 1998. IPF2alpha-I: an index of lipid peroxidation in humans. Proc. Natl. Acad. Sci. U. S. A 95, 3449-3454.

Rebec, G.V., Pierce, R.C. 1994. A vitamin as neuromodulator: ascorbate release into the extracellular fluid of the brain regulates dopaminergic and glutamatergic transmission. Prog. Neurobiol. 43, 537-565.

Rice, M.E., Russo-Menna, I. 1998. Differential compartmentalization of brain ascorbate and glutathione between neurons and glia. Neuroscience 82, 1213-1223.

Rogers, J., Kirby, L.C., Hempelman, S.R., Berry, D.L., McGeer, P.L., Kaszniak, A.W., Zalinski, J., Cofield, M., Mansukhani, L., Wilson, P. 1993. Clinical trial of indomethacin in Alzheimer's disease.

Neurology 43, 1609-1611.

Rothwell, N.J., Luheshi, G.N. 2000. Interleukin 1 in the brain: biology, pathology and therapeutic target. Trends Neurosci. 23, 618-625.

Sanchez-Moreno, C., Paniagua, M., Madrid, A., Martin, A. 2003. Protective effect of vitamin C against the ethanol mediated toxic effects on human brain glial cells. J. Nutr. Biochem. 14, 606-613.

Seibert, K., Zhang, Y., Leahy, K., Hauser, S., Masferrer, J., Isakson, P. 1997. Distribution of COX-1 and COX-2 in normal and inflamed tissues. Adv. Exp. Med. Biol. 400A, 167-170.

Smith, W.L., DeWitt, D.L., Garavito, R.M. 2000. Cyclooxygenases: structural, cellular, and molecular biology. Annu. Rev. Biochem. 69, 145-182.

Smith, W.L., Marnett, L.J. 1991. Prostaglandin endoperoxide synthase: structure and catalysis. Biochim. Biophys. Acta 1083, 1-17.

Svensson, L., Wu, C., Hulthe, P., Johannessen, K., Engel, J.A. 1993. Effect of ageing on extracellular ascorbate concentration in rat brain. Brain Res. 609, 36-40.

Takeguchi, C., Sih, C.J. 1972. A rapid spectrophotometric assay for prostaglandin synthetase: application to the study of non-steroidal antiinflammatory agents. Prostaglandins 2, 169-184.

Touzani, O., Boutin, H., LeFeuvre, R., Parker, L., Miller, A., Luheshi, G., Rothwell, N. 2002. Interleukin-1 influences ischemic brain damage in the mouse independently of the interleukin-1 type I receptor. J. Neurosci. 22, 38-43.

Tyurin, V.A., Tyurina, Y.Y., Borisenko, G.G., Sokolova, T.V., Ritov, V.B., Quinn, P.J., Rose, M., Kochanek, P., Graham, S.H., Kagan, V.E. 2000. Oxidative stress following traumatic brain injury in rats:

quantitation of biomarkers and detection of free radical intermediates. J. Neurochem. 75, 2178-2189.

Vanderhoek, J.Y., Lands, W.E. 1973. The inhibition of the fatty acid oxygenase of sheep vesicular gland by antioxidants. Biochim. Biophys. Acta 296, 382-385.

Vane, J.R., Bakhle, Y.S., Botting, R.M. 1998. Cyclooxygenases 1 and 2. Annual Review of Pharmacology & Toxicology 38, 97-120.

Wells, W.W., Xu, D.P. 1994. Dehydroascorbate reduction. J. Bioenerg. Biomembr. 26, 369-377.

Winkler, B.S., Orselli, S.M., Rex, T.S. 1994. The redox couple between glutathione and ascorbic acid: a chemical and physiological perspective. Free Radic. Biol. Med. 17, 333-349.

Xiao, G., Tsai, A.L., Palmer, G., Boyar, W.C., Marshall, P.J., Kulmacz, R.J. 1997. Analysis of hydroperoxide-induced tyrosyl radicals and lipoxygenase activity in aspirin-treated human

prostaglandin H synthase-2. Biochemistry 36, 1836-1845.

Yamagata, K., Andreasson, K.I., Kaufmann, W.E., Barnes, C.A., Worley, P.F. 1993. Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 11, 371-386.


Repository Staff Only: item control page