Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10

Takehito Suzuki, Mayuko Uchida, Yuji Takeda, Chiemi Mori, Atsushi Onuki, Yoko Miyazaki, Ken Onda, Setsuo Ushikoshi, Kotaro Shitori, Kazuaki Tanaka, Hidetoshi Morita, Tatsuya Takizawa

Research output: Contribution to journalArticle

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 p22phox 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.

Original languageEnglish
Pages (from-to)1384-1389
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume77
Issue number7
DOIs
Publication statusPublished - 2013
Externally publishedYes

Fingerprint

Rhizopus
NF-kappa B
Rats
Hepatocytes
Nitric Oxide
Cells
Cell Line
Superoxides
RNA-Directed DNA Polymerase
Nitric Oxide Synthase Type II
Metabolites
Nitrites
Reverse Transcriptase Polymerase Chain Reaction
NADP
Ammonium Compounds
Cell culture
Gene expression
Culture Media
Lipopolysaccharides
Real-Time Polymerase Chain Reaction

Keywords

  • Ammonium pyrrolidine-1-carbodithioate (APDC)
  • Hepatocyte cell line
  • Nitric oxide (NO)
  • Nuclear factor-κB
  • Rhizopus oryzae

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

Cite this

Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10. / Suzuki, Takehito; Uchida, Mayuko; Takeda, Yuji; Mori, Chiemi; Onuki, Atsushi; Miyazaki, Yoko; Onda, Ken; Ushikoshi, Setsuo; Shitori, Kotaro; Tanaka, Kazuaki; Morita, Hidetoshi; Takizawa, Tatsuya.

In: Bioscience, Biotechnology and Biochemistry, Vol. 77, No. 7, 2013, p. 1384-1389.

Research output: Contribution to journalArticle

Suzuki, T, Uchida, M, Takeda, Y, Mori, C, Onuki, A, Miyazaki, Y, Onda, K, Ushikoshi, S, Shitori, K, Tanaka, K, Morita, H & Takizawa, T 2013, 'Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10', Bioscience, Biotechnology and Biochemistry, vol. 77, no. 7, pp. 1384-1389. https://doi.org/10.1271/bbb.120951
Suzuki, Takehito ; Uchida, Mayuko ; Takeda, Yuji ; Mori, Chiemi ; Onuki, Atsushi ; Miyazaki, Yoko ; Onda, Ken ; Ushikoshi, Setsuo ; Shitori, Kotaro ; Tanaka, Kazuaki ; Morita, Hidetoshi ; Takizawa, Tatsuya. / Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10. In: Bioscience, Biotechnology and Biochemistry. 2013 ; Vol. 77, No. 7. pp. 1384-1389.
@article{da0ac87c25e2487889399c3b7a01075d,
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 p22phox 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.",
keywords = "Ammonium pyrrolidine-1-carbodithioate (APDC), Hepatocyte cell line, Nitric oxide (NO), Nuclear factor-κB, Rhizopus oryzae",
author = "Takehito Suzuki and Mayuko Uchida and Yuji Takeda and Chiemi Mori and Atsushi Onuki and Yoko Miyazaki and Ken Onda and Setsuo Ushikoshi and Kotaro Shitori and Kazuaki Tanaka and Hidetoshi Morita and Tatsuya Takizawa",
year = "2013",
doi = "10.1271/bbb.120951",
language = "English",
volume = "77",
pages = "1384--1389",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "7",

}

TY - JOUR

T1 - Aqueous extracts of Rhizopus oryzae induced nitric oxide production in rat hepatocyte cell line RLN-10

AU - Suzuki, Takehito

AU - Uchida, Mayuko

AU - Takeda, Yuji

AU - Mori, Chiemi

AU - Onuki, Atsushi

AU - Miyazaki, Yoko

AU - Onda, Ken

AU - Ushikoshi, Setsuo

AU - Shitori, Kotaro

AU - Tanaka, Kazuaki

AU - Morita, Hidetoshi

AU - Takizawa, Tatsuya

PY - 2013

Y1 - 2013

N2 - 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 p22phox 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.

AB - 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 p22phox 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.

KW - Ammonium pyrrolidine-1-carbodithioate (APDC)

KW - Hepatocyte cell line

KW - Nitric oxide (NO)

KW - Nuclear factor-κB

KW - Rhizopus oryzae

UR - http://www.scopus.com/inward/record.url?scp=84882772475&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84882772475&partnerID=8YFLogxK

U2 - 10.1271/bbb.120951

DO - 10.1271/bbb.120951

M3 - Article

AN - SCOPUS:84882772475

VL - 77

SP - 1384

EP - 1389

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 7

ER -