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Inducible nitric oxide synthase (iNOS) has been detected in a number of pathologic conditions in the central nervous system. This study was investigated the patterns of iNOS expression in the neuronal PC12 cell and the effects of nitric oxide on the apoptosis of PC12 cells. Methods : The stimulating agents for induction of iNOS expression in PC12 cells were bacterial lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-α), and interferon- gamma (IFN-γ). Results : The expression iNOS mRNA and protein in PC12 cells stimulated with LPS/TNF-α/IFN-γ were profoundly increased. The expression of iNOS mRNA arose at 6 hours, peaked at 12 hours, and declined to 48 hours after LPS/TNF-α/IFN-γ treatment. iNOS protein was increased up to 24 hours in LPS/TNF-α/IFN-γ treated PC12 cells while the expression of nNOS was unaffected. Accumulation of NO derivatives in the culture media was markedly increased at least at up to 48 hours after LPS/TNF-α/IFNtreatment. The induction of iNOS expression and NO production in differentiated PC12 cells was correlated with apoptotic cell death judged by transmission electron microscopy and DNA fragmentation from the results of the Terminal deoxynucleotidyl-transferase-mediated dUDP biotin nick end-labeling (TUNEL) method. After treatment with NOS inhibitor, Nmonomethylarginine (NMMA), a profound decrease in NO production by LPS/TNF-α/IFN-γ treated PC12 cells was noted. And the LPS/TNF-α/IFN-γ induced apoptosis was prevented by the NMMA treatment. Conclusions : From the above results it is concluded that the expression of iNOS in differentiated PC12 cells is induced by the combined application of LPS, TNF-α, and IFN-γ . And the apoptosis of cultured PC12 cells is mediated by iNOSderived NO.

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