Open AccessNeuroprotective effects of cyclooxygenase‐2 inhibitor celecoxib against toxicity of LPS‐stimulated macrophages toward motor neurons
Author(s) -
HUANG Yong,
LIU Jing,
WANG Lizhen,
ZHANG Weiyu,
ZHU Xingzu
Publication year - 2005
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2005.00136.x
Subject(s) - lipopolysaccharide , nitric oxide , prostaglandin e2 , nitric oxide synthase , chemistry , cyclooxygenase , tumor necrosis factor alpha , neuroprotection , microbiology and biotechnology , celecoxib , macrophage , viability assay , prostaglandin e , pharmacology , biochemistry , in vitro , biology , enzyme , immunology , endocrinology , organic chemistry
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 E 2 (PGE 2 ), 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 inLPS‐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.