title: Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10.
abstract: Aqueous extracts of Rhizopus oryzae (Aq-ROU) have a broad range of physiological activity. Here we identified a new physiological effect of Aq-ROU in rat hepatocyte cell line RLN-10. Aq-ROU induced the accumulation of nitrite, a stable metabolite nitric oxide (NO), in cell culture medium and induced potent diaminofluorescein-FM diacetate staining in the cells. Real-time reverse transcriptase (RT)-PCR analysis showed marked inducible NO synthase gene expression. Additionally, markedly enhanced expression of p22(phox) and temporally increased expression of NADPH oxidase1 indicated that superoxide was produced. Nuclear translocation of nuclear factor-kappa (NF-κ) B p65 increased remarkably following Aq-ROU and following lipopolysaccharide treatment, a potent activator of NF-κB. Ammonium pyrrolidine-1-carbodithioate, an inhibitor of NF-κB, inhibited NO production following Aq-ROU treatment. Our data indicate that Aq-ROU induces NO production and potentially the production of superoxide, which may contribute to the broad range of physiological effects observed for Aq-ROU ingested by animals.
authors.affiliation: Graduate School of Veterinary Medicine, Azabu University, Sagamihara 252-5201, Japan.
authors.forename: Takehito
authors.initials: T
authors.lastname: Suzuki
authors.forename: Mayuko
authors.initials: M
authors.lastname: Uchida
authors.forename: Yuji
authors.initials: Y
authors.lastname: Takeda
authors.forename: Chiemi
authors.initials: C
authors.lastname: Mori
authors.forename: Atsushi
authors.initials: A
authors.lastname: Onuki
authors.forename: Yoko
authors.initials: Y
authors.lastname: Miyazaki
authors.forename: Ken
authors.initials: K
authors.lastname: Onda
authors.forename: Setsuo
authors.initials: S
authors.lastname: Ushikoshi
authors.forename: Kotaro
authors.initials: K
authors.lastname: Shitori
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Tanaka
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Morita
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Takizawa
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases / Q000235:genetics; D009569:Nitric Oxide / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics; D011759:Pyrrolidines / Q000494:pharmacology; D051381:Rats; D012233:Rhizopus / Q000737:chemistry*; D013481:Superoxides / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014867:Water / Q000737:chemistry*
chemical_list: D011759:Pyrrolidines; D051996:Transcription Factor RelA; D014867:Water; D013481:Superoxides; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D009247:NADH, NADPH Oxidoreductases; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases; C515768:Cyba protein, rat