Neuroprotective effects of cyclooxygenase-2 inhibitor celecoxib against toxicity of LPS-stimulated macrophages toward motor neurons

Acta Pharmacol Sin. 2005 Aug;26(8):952-8. doi: 10.1111/j.1745-7254.2005.00136.x.


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-gamma (IFN-gamma) 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-gamma for 12 h or 24 h, the release of prostaglandin E2 (PGE2), nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) 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-alpha and IL-1beta 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-alpha and IL-1beta from macrophages were attenuated by pre-treatment with celecoxib. However, celecoxib showed no effect on the ROS levels upregulated by LPS+IFN-gamma in the macrophage supernatant. The mRNA levels of COX-2, iNOS, TNF-alpha and IL-1beta 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.

MeSH terms

  • Animals
  • Celecoxib
  • Cell Line
  • Cell Survival / drug effects
  • Cells, Cultured
  • Coculture Techniques
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 Inhibitors / pharmacology*
  • Dinoprostone / metabolism
  • Dose-Response Relationship, Drug
  • Female
  • Interferon-gamma / pharmacology
  • Interleukin-1 / genetics
  • Lipopolysaccharides / toxicity*
  • Macrophage Activation / drug effects
  • Macrophages, Peritoneal / cytology
  • Macrophages, Peritoneal / drug effects*
  • Macrophages, Peritoneal / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Motor Neurons / cytology
  • Motor Neurons / drug effects*
  • Neuroprotective Agents / pharmacology
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type II / genetics
  • Pyrazoles / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Radioimmunoassay
  • Reactive Oxygen Species / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sulfonamides / pharmacology*
  • Time Factors
  • Tumor Necrosis Factor-alpha / genetics


  • Cyclooxygenase 2 Inhibitors
  • Interleukin-1
  • Lipopolysaccharides
  • Neuroprotective Agents
  • Pyrazoles
  • RNA, Messenger
  • Reactive Oxygen Species
  • Sulfonamides
  • Tumor Necrosis Factor-alpha
  • Nitric Oxide
  • Interferon-gamma
  • Nitric Oxide Synthase Type II
  • Cyclooxygenase 2
  • Celecoxib
  • Dinoprostone