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

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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.

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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

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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.

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Authors and Affiliations

  1. Department of Biomedical Sciences, College of Life Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan

    Yuko Yamauchi, Tetsuya Okuyama, Toshinari Ishii & Mikio Nishizawa

  2. Research Organization of Science and Technology, Ritsumeikan University, Kusatsu, Shiga, Japan

    Tadayoshi Okumura

  3. Department of Surgery, Kansai Medical University, Hirakata, Osaka, Japan

    Tadayoshi Okumura

  4. Center for Supporting Pharmaceutical Education, Daiichi University of Pharmacy, 22-1 Tamagawa-cho, Minami-ku, Fukuoka, 815-8511, Japan

    Yukinobu Ikeya

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  1. Yuko Yamauchi
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  2. Tetsuya Okuyama
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  3. Toshinari Ishii
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  4. Tadayoshi Okumura
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  5. Yukinobu Ikeya
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Correspondence to Yukinobu Ikeya.

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

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Yamauchi, Y., Okuyama, T., Ishii, T. et al. Sakuranetin downregulates inducible nitric oxide synthase expression by affecting interleukin-1 receptor and CCAAT/enhancer-binding protein β. J Nat Med 73, 353–368 (2019). https://doi.org/10.1007/s11418-018-1267-x

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  • DOI: https://doi.org/10.1007/s11418-018-1267-x

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