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Journal of Natural Medicines

, Volume 72, Issue 1, pp 181–191 | Cite as

Synergistic effect of baicalein, wogonin and oroxylin A mixture: multistep inhibition of the NF-κB signalling pathway contributes to an anti-inflammatory effect of Scutellaria root flavonoids

  • Tomofumi Shimizu
  • Nobuhiko Shibuya
  • Yuji Narukawa
  • Naohiro Oshima
  • Noriyasu Hada
  • Fumiyuki KiuchiEmail author
Original Paper

Abstract

Scutellaria root, the root of Scutellaria baicalensis Georgi, is a crude drug used for inflammatory diseases. In our previous report, the combination of flavonoids contained in Scutellaria root have been found to inhibit PGE2 production more strongly than individual flavonoids. Here, to investigate the mechanism of the synergistic effect, we examined the effects of an equimolar mixture (F-mix) of baicalein (1), wogonin (2) and oroxylin A (3) on the production of PGE2 in LPS-treated J774.1 cells. Although 1 and 3 inhibited COX-2 activity, the F-mix showed no synergistic effect on COX-2 inhibition. Therefore, we investigated the steps leading to the activation of COX-2 protein. Compounds 13 and F-mix inhibited the expression of COX-2 protein. However, only 2 inhibited the expression of COX-2 mRNA among the flavonoids, and the F-mix showed no synergistic effect. Only 1 inhibited NF-κB translocation into the nucleus, and the F-mix showed no synergistic effect. Although 2 did not affect NF-κB translocation, it strongly inhibited NF-κB-dependent transcriptional activity, and the F-mix inhibited the activity slightly more than 2. Compounds 13 also inhibited NO production, and the F-mix showed a synergistic effect. However, the effects of each flavonoid on the expression of iNOS mRNA were not consistent with those on COX-2 mRNA. Because the flavonoids inhibit different steps in the production of PGE2 and NO, and their mixture did not show apparent synergistic effects in each step, we conclude that the synergistic effect of the flavonoid mixture reflects the total effect of the flavonoids inhibiting different steps in the NF-κB signalling pathway.

Keywords

Scutellaria baicalensis Flavonoid Synergistic effect Prostaglandin E2 Inflammation Nuclear factor-kappa B 

Abbreviations

COX-2

Cyclooxygenase-2

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Foetal bovine serum

GAPDH

Glyceraldehyde 3-phosphate dehydrogenase

IFN-γ

Interferon gamma

iNOS

Inducible nitric oxide synthase

IRAK

Interleukin-1 receptor-associated kinase

IRF-3

Interferon regulatory factor 3

IκB

Inhibitor of nuclear factor-kappa B

JAK

Janus kinase

l-NMMA

N G-Monomethyl-l-arginine acetate

LPS

Lipopolysaccharide

MAPK

Mitogen-activated protein kinase

MTT

3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide

MyD88

Myeloid differentiation primary response gene 88

NF-κB

Nuclear factor-kappa B

NO

Nitric oxide

OPLS

Orthogonal projection to latent structure

PBS

Phosphate buffered saline

PCA

Principal component analysis

PG

Prostaglandin

PVDF

Polyvinylidene difluoride

STAT

Signal transducer and activator of transcription

TAK1

Transforming growth factor beta-activated kinase 1

TGF-β

Transforming growth factor beta

TLR

Toll-like receptor

TNF-α

Tumour necrosis factor alpha

TRAF6

Tumour necrosis factor receptor-associated factor 6

Notes

Acknowledgements

This work was supported by JSPS KAKENHI grant number JP26460130, Keio Gijuku Academic Development Funds and MEXT-Supported Program for the Strategic Research Foundation at Private Universities. The authors thank Dr. Megumi Funakoshi-Tago for the valuable discussions.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

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Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK 2017

Authors and Affiliations

  1. 1.Faculty of PharmacyKeio UniversityTokyoJapan
  2. 2.Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan

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