Original Article

Exp Neurobiol 2011; 20(2): 100-109

Published online June 30, 2011

© The Korean Society for Brain and Neural Sciences

Protective Effect of Coriolus versicolor Cultivated in Citrus Extract Against Nitric Oxide-Induced Apoptosis in Human Neuroblastoma SK-N-MC Cells

Byung-Chul Kim1, Youn-Sub Kim1, Jin-Woo Lee2, Jin-Hee Seo2, Eun-Sang Ji 2, Hyejung Lee3, Yong-Il Park4 and Chang-Ju Kim2*

1Department of Anatomy-Pointology, College of Oriental Medicine, Kyungwon University, Seongnam 461-701, 2Department of Physiology, College of Medicine, Kyung Hee University, Seoul 130-701, 3Acupuncture and Meridian Science Research Center, Kyung Hee University, Seoul 130-701, 4Division of Biotechnology and Collaborative Institute of Science and Technology, The Catholic University of Korea, Bucheon 420-717, Korea

Correspondence to: *To whom correspondence should be addressed.
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Nitric oxide (NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive NO is believed to be a mediator of neurotoxicity. The medicinal plant Coriolus versicolor is known to possess anti-tumor and immune-potentiating activities. In this study, we investigated whether Coriolus versicolor possesses a protective effect against NO donor sodium nitroprusside (SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. We utilized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-MC cells. MTT assay showed that SNP treatment significantly reduces the viability of cells, and the viabilities of cells pre-treated with the aqueous extract of Coriolus versicolor cultivated in citrus extract (CVEcitrus) was increased. However, aqueous extract of Coriolus versicolor cultivated in synthetic medium (CVEsynthetic) showed no protective effect and aqueous citrus extract (CE) had a little protective effect. The cell treated with SNP exhibited several apoptotic features, while those pre-treated for 1 h with CVEcitrus prior to SNP expose showed reduced apoptotic features. The cells pre-treated for 1 h with CVEcitrus prior to SNP expose inhibited p53 and Bax expressions and caspase-3 enzyme activity up-regulated by SNP. We showed that CVEcitrus exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells. Our study suggests that CVEcitrus has therapeutic value in the treatment of a variety of NO-induced brain diseases.

Keywords: Coriolus versicolor, nitric oxide, apoptosis, human neuroblastoma