Abstract
Aim:
To establish an in vitro injured motor neuronal model and investigate the neuroprotective effects and possible mechanism of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on this model.
Methods:
After macrophages were stimulated with lipopolysaccharide (LPS)+interferon-γ (IFN-γ) in the presence or absence of celecoxib for 24 h, the cell-free supernatant of LPS-stimulated macrophages was transferred to the culture of NSC34 cells. Viability of NSC34 cells was assessed by MTT assay after a further 24 h and 72 h incubation. After macrophages were stimulated by LPS+IFN-γ for 12 h or 24 h, the release of prostaglandin E2 (PGE2), nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) from macrophages was measured by radioimmunoassay, Griess assay, fluorescence assay and enzyme-linked immunosorbent assay, respectively. The mRNA levels of COX-2, inducible nitric oxide synthase (iNOS), TNF-α and IL-1β in macrophages were determined by reverse transcription-polymerase chain reaction after macrophages were stimulated for 6 h and 12 h.
Results:
The supernatant of LPS-stimulated mouse macrophages induced the death of NSC34 cells and celecoxib protected the NSC34 cells against this toxicity. The LPS-induced increases in the release of PGE2, NO, TNF-α and IL-1β from macrophages were attenuated by pre-treatment with celecoxib. However, celecoxib showed no effect on the ROS levels upregulated by LPS+IFN-γ in the macrophage supernatant. The mRNA levels of COX-2, iNOS, TNF-α and IL-1β were increased in LPS-activated macrophages and, except COX-2, reduced by pre-treatment with celecoxib.
Conclusion:
An in vitro injured motor neuronal model was established by using the toxicity of LPS-stimulated mouse macrophages toward motor neuronal NSC34 cells. In this model, celecoxib exerted neuroprotective effects on motor neurons via an inhibition of the neurotoxic secretions from activated macrophages.
Similar content being viewed by others
Article PDF
References
Dib M . Amyotrophic lateral sclerosis progress and prospects for treatment. Drugs 2003; 63: 289–310.
McGeer PL, McGeer EG . Inflammatory processes in amyotrophic lateral sclerosis. Muscle Nerve 2002; 26: 459–70.
Alexianu ME, Kozovska ME, Appel SH . Immune reactivity in a mouse model of familial ALS correlates with disease progression. Neurology 2001; 57: 1282–9.
Kawamata T, Akiyama H, Yamada T, McGeer PL . Immunologic reactions in amyotrophic lateral sclerosis brain and spinal cord tissue. Am J Pathol 1992; 140: 691–707.
Troost D, Van den Oord JJ, Vianney de Jong JM . Immunohistochemical characterization of the inflammatory infiltrate in amyotrophic lateral sclerosis. Neuropathol Appl Neurobiol 1990; 16: 401–10.
Abe K, Abe Y, Saito H . Agmatine suppresses nitric oxide production in microglia. Brain Res 2000; 872: 141–8.
Piani D, Frei K, Do KQ, Cuenod M, Fontana A . Murine brain macrophages induce NMDA receptor mediated neurotoxicity in vitro by secreting glutamate. Neurosci Lett 1991; 133: 159–62.
Chao CC, Hu S, Peterson PK . Glia, cytokines, and neurotoxicity. Crit Rev Neurobiol 1995; 9: 189–205.
Levi G, Minghetti L, Aloisi F . Regulation of prostanoid synthesis in microglial cells and effects of prostacyclin E2 on microglial functions. Biochimie 1998; 80: 899–904.
Almer G, Guegan C, Teismann P, Naini A, Rosoklija G, Hays AP, et al. Increased expression of the pro–inflammatory enzyme cyclooxygenase–2 in amyotrophic lateral sclerosis. Ann Neurol 2001; 49: 176–85.
Yasojima K, Tourtellotte WW, McGeer EG, McGeer PL . Marked increase in cyclooxygenase-2 in ALS spinal cord: implications for therapy. Neurology 2001; 576: 952–6.
Maihofner C, Probst-Cousin S, Bergmann M, Neuhuber W, Neundorfer B, Heuss D . Expression and localization of cyclooxygenase-1 and -2 in human sporadic amyotrophic lateral sclerosis. Eur J Neurosci 2003; 18: 1527–34.
Drachman DB, Rothstein JD . Inhibition of cyclooxygenase-2 protects motor neurons in an organotypic model of amyotrophic lateral sclerosis. Ann Neurol 2000; 48: 792–5.
Seibert K, Zhang Y, Leahy K, Hauser S, Masferrer J, Perkins W, et al. Pharmacological and biochemical demonstration of the role of cyclooxygenase-2 in inflammation and pain. Proc Natl Acad Sci USA 1994; 91: 12013–7.
Drachman DB, Frank K, Dykes-Hoberg M, Teismann P, Almer G, Przedborski S, et al. Cyclooxygenase 2 inhibition protects motor neurons and prolongs survival in a transgenic mouse model of ALS. Ann Neurol 2002; 52: 771–8.
Thomas WE . Brain marcrophages: evaluation of microglia and their functions. Brain Res Brain Res Rev 1992; 17: 61–74.
Portegies P, Cohen ES . Possible etiological role retroviruses and enteroviruses in the development of amyotrophic lateral sclerosis. Ned Tijdschr Geneeskd 2002; 146: 1398–400.
Cashman NR, Durham HD, Blusztajn JK, Oda K, Tabira T, Shaw IT, et al. Neuroblastoma X spinal cord (NSC) hybrid cell lines resemble developing motor neurons. Dev Dyn 1992; 194: 209–21.
He BP, Wen WY, Strong MJ . Activated microglia (BV-2) facilitation of TNF-α-mediated motor neuron death in vitro. J Neuroimmunol 2002; 128: 31–8.
Klegeris A, McGeer PL . Cyclooxygenase and 5–lipoxygenase inhibitors protect against mononuclear phagocyte neurotoxicity. Neurobiol Aging 2002; 237: 787–94.
Hansen MB, Nielsen SE, Berg K . Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Meth 1989; 119: 203–10.
Eigler A, Moeller J, Endres S . Exogenous and endogenous nitric oxide attenuates tumor necrosis factor synthesis in ht murine macrophage cell line RAW 264.7. J Immunol 1995; 154: 4048–54.
Gunasekar PG, Borowitz JL, Isom GE . Cyanide-induced generation of oxidative species: involvement of nitric oxide synthase and cyclooxygenase-2. J Pharmacol Exp Ther 1998; 285: 236–41.
Hewett SJ, Uliasz TF, Vidwans AS, Hewett JA . Cyclooxygenase-2 contributes to N-methyl-D-aspartate-mediated neuronal cell death in primary cortical cell culture. J Pharmacol Exp Ther 2000; 293: 417–25.
Jeon HK, Jung NP, Choi IH, Oh YK, Shin HC, Gwag BJ . Substance P augments nitric oxide production and gene expression in murine macrophages. Immunopharmacology 1999; 41: 219–26.
Suzuki T, Ogata A, Tashiro K, Nagashima K, Tamura M, Yasui K, et al. Japanese encephalitis virus up-regulates expression of macrophage migration inhibitory factor (MIF) mRNA in the mouse brain. Biochim Biophys Acta 2000; 1517: 100–6.
Choi CY, Kim JY, Kim YS, Chung YC, Hahm KS, Jeong HG . Augmentation of macrophage functions by an aqueous extract isolated from Platycodon grandiflorum. Can Lett 2001; 166: 17–25.
Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔC T method. Methods 2001; 25: 402–8.
Klegeris A, Walker DG, McGeer PL . Toxicity of human THP-1 monocytic cells towards neuron-like cells is reduced by non-steroidal anti-inflammatory drugs (NSAIDS). Neuropharmacology 1999; 38: 1017–25.
Callejas NA, Castrillo A, Bosca L, Marin-Sanz P . Inhibition of prostaglandin synthesis up-regulates cyclooxygenase-2 induced by lipopolysaccharide and peroxisomal proliferators. J Pharmacol Exp Ther 1999; 288: 1235–41.
Nathan C . Nitric oxide as a secretory product of mammalian cells. FASEB J 1992; 6: 3051–64.
Heneka MT, Feinstein DL . Expression and function of inducible nitric oxide synthase in neurons. J Neuroimmunol 2001; 114: 8–18.
Kim EJ, Kwon KJ, Park JY, Lee SH, Moon CH, Baik EJ . Neuroprotective effects of prostaglandin E2 or cAMP against microglia and neuronal free radical mediated toxicity associated with inflammation. J Neurosci Res 2002: 70: 97–107.
Wang T, Qin L, Liu B, Liu Y, Wilson B, Eling TE, et al. Role of reactive oxygen species in LPS-induced production of prostaglandin E2 in microglia. J Neurochem 2004; 88: 939–47.
Klegeris A, McGeer PL . Inflammatory cytokine levels are influenced by interactions between THP-1 monocytic, U-373 MG astrocytic, and SH-SY5Y neuronal cell lines of human origin. Neurosci Lett 2001; 31: 341–4.
Casolini P, Catalani A, Zuena AR, Angelucci L . Inhibition of COX-2 reduces the age-dependent increase of hippocampal inflammatory markers, corticosterone secretion, and behavioral impairments in the rat. J Neurosci Res 2002; 68: 337–43.
Chou SM, Wang HS, Komai K . Colocalization of NOS and SOD1 in neurofilament accumulation within motor neurons of amyotrophic lateral sclerosis: an immunohistochemical study. J Chem Neuroanat 1996; 10: 249.
Sasaki S, Shibata N, Komori T, Iwata M . iNOS and nitro tyrosine immunoreactivity in amyotrophic lateral sclerosis. Neurosci Lett 2000; 291: 44.
Almer G, Vukosavic S, Romero N, Przedborski S . Inducible nitric oxide synthase upregulation in a transgenic mouse model of familial amyotrophic lateral sclerosis. J Neurochem 1999; 72: 2415–25.
Poloni M, Facchetti D, Mai R, Micheli A, Agnoletti L, Francolini G, et al. Circulating levels of tumour necrosis factor-alpha and its soluble receptors are increased in the blood of patients with amyotrophic lateral sclerosis. Neurosci Lett 2000; 287: 211–4.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Huang, Y., Liu, J., Wang, Lz. et al. Neuroprotective effects of cyclooxygenase-2 inhibitor celecoxib against toxicity of LPS-stimulated macrophages toward motor neurons. Acta Pharmacol Sin 26, 952–958 (2005). https://doi.org/10.1111/j.1745-7254.2005.00136.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2005.00136.x
Keywords
This article is cited by
-
Celecoxib Inhibits Prion Protein 90-231-Mediated Pro-inflammatory Responses in Microglial Cells
Molecular Neurobiology (2016)
-
COX‐2‐mediated Inflammation in Fat Is Crucial for Obesity‐linked Insulin Resistance and Fatty Liver
Obesity (2009)
-
Neuronal Cyclooxygenase-2 Activity and Prostaglandins PGE2, PGD2, and PGF2α Exacerbate Hypoxic Neuronal Injury in Neuron-enriched Primary Culture
Neurochemical Research (2008)