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Sakuranetin downregulates inducible nitric oxide synthase expression by affecting interleukin-1 receptor and CCAAT/enhancer-binding protein β

  • Yuko Yamauchi
  • Tetsuya Okuyama
  • Toshinari Ishii
  • Tadayoshi Okumura
  • Yukinobu Ikeya
  • Mikio Nishizawa
Original Paper
  • 77 Downloads

Abstract

Pruni Cortex is a herbal drug from the bark of the Japanese flowering cherries, Prunus jamasakura or Prunus verecunda, and is included in the traditional Japanese herbal (Kampo) formula Jumihaidokuto, which is administered orally to patients suffering from inflammatory skin diseases. The flavanones contained in Pruni Cortex (e.g., sakuranetin and naringenin) have potent anti-inflammatory, anti-allergic, and anti-microbial activities. Although the effects of Pruni Cortex on skin disease have been well studied, reports regarding its pharmacological effects on the liver are limited. In this study, we extracted the bark of Prunus jamasakura and purified it to isolate the pharmacologically active constituents by monitoring nitric oxide (NO) production in rat hepatocytes that were treated with the pro-inflammatory cytokine, interleukin (IL)-1β. Sakuranetin and (−)-naringenin, which were present in an ethyl acetate-soluble fraction of the bark extract, significantly inhibited NO induction and inducible nitric oxide synthase (iNOS) expression. These two flavanones decreased the expression of type 1 IL-1 receptor gene and phosphorylation of Akt, also known as protein kinase B, which is regulated by phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K). Furthermore, sakuranetin decreased the phosphorylation of the activator isoforms of CCAAT/enhancer-binding protein β (C/EBPβ), which synergistically activates the transcription of the iNOS gene with nuclear factor κB (NF-κB). Therefore, sakuranetin inhibited the co-activating activity of C/EBPβ with NF-κB, leading to the suppression of iNOS gene expression in hepatocytes. Taken together, sakuranetin in Pruni Cortex downregulated the iNOS gene by inhibiting PI3K/Akt signal transduction and the phosphorylation of C/EBPβ. These results imply that sakuranetin may be primarily responsible for the anti-inflammatory effects of Pruni Cortex in the liver.

Keywords

Cherry bark Nitric oxide Kampo medicine Interleukin 1 receptor CCAAT/enhancer-binding protein β Nuclear factor κB 

Abbreviations

PI3K

Phosphatidylinositol-3-kinase

NF-κB

Nuclear factor-κB

C/EBPβ

CCAAT/enhancer-binding protein β

NO

Nitric oxide

IL

Interleukin

iNOS

Inducible nitric oxide synthase

IL1R1

Interleukin 1 receptor, type 1

NMR

Nuclear magnetic resonance

qRT-PCR

Quantitative reverse transcription–polymerase chain reaction

EF

Elongation factor 1α

AABS

A activator-binding site

CMV

Cytomegalovirus

PC

Pruni Cortex

asRNA

Antisense transcript

LAP

Liver-enriched activator protein

Notes

Acknowledgements

We thank Dr. Yuji Hasegawa for mass spectra analyses, Ms. Yuki Nakano for her technical assistance, and Ms. Noriko Kanazawa for her secretarial assistance. This work was supported in part by the Asia-Japan Research Institute of Ritsumeikan Asia-Japan Research Organization, Ritsumeikan University.

Compliance with ethical standards

Conflict of interest

Y. Yamauchi and T. Ishii performed this study as graduate students of the Graduate School of Life Sciences, Ritsumeikan University.

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Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biomedical Sciences, College of Life SciencesRitsumeikan UniversityKusatsuJapan
  2. 2.Research Organization of Science and TechnologyRitsumeikan UniversityKusatsuJapan
  3. 3.Department of SurgeryKansai Medical UniversityHirakataJapan
  4. 4.Center for Supporting Pharmaceutical EducationDaiichi University of PharmacyFukuokaJapan

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