ENHANCEMENT OF NITRIC OXIDE SYNTHASE GENE EXPRESSION BY OLANZAPINE.

초록

Statement of the Study : Nitric oxide(NO) is considered as one of the major intercellular messengers in the brain. The purpose of this study is to investigate how olanzapine, an atypical antipsychotic, is related to the NO synthesis mechanism. Methods: Olanzapine was applied on the protein expression of neuronal nitric oxide synthase(nNOS) and brain derived neurotrophic factor(BDNF) was determined by Western-blot analysis. Luciferase reporter gene system and the activities were transfected to the human dopaminergic cell line(SH-SY5Y), and then their activities were determined by luminometer. Nuclear complex were extracted from mouse dopaminergic cell line(CATH.A), peripheral neuronal cell line(PC12), and SH-SY5Y, and then identified by electromobility shift assay(EMSA). Multiple DNA-protein complexes in CRE site of hyman nNos exon. If basal promoter and effects of three different protein kinase inhibitors on the complex-DNA binding activities in SH-SY5Y cells were assayed by using EMSA. Summary of Results: nNOS expression was increased in olanzapine group at low concentration(0.1μΜ, 1μΜ) significantly. The relative luciferase activities were dependent on the cyclic AMP response element(CRE) and nuclear factor kappa B-like activator protein(NFkB-like AP2) consensus sequences. Regardless of(GT)17A(TG)13 repeated sequences, the activities were more sensitive in NFkB-like AP2 site and CRE site. Conclusion: The results of this study imply that olanzapine increases NO synthesis, and increased NO synthesis may affect decrease in dopamine efflux. Two major transcriptional factors(CREB and NFkB-lie AP2) in 5-flanking region of human nNOS gene play an important role in NO synthesis. This suggests that CRE site can be influenced by olanzapine.