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In Vitro Anti-inflammatory and Antioxidant Activities of 3,5,4′-Trihydroxy-6,7-Methylenedioxyflavone-O-Glycosides and Their Aglycone from Leaves of Polygonum tinctorium Lour

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Abstract

Polygonum tinctorium Lour (indigo plant) has been regarded as a useful medicinal plant for traditional herbal medicine. The polyphenolic fraction of indigo leaves exhibited anti-inflammatory activities as determined by the suppressed synthesis of nitric oxide (NO) in cultured RAW264 macrophage cells. The acid hydrolysate of the fraction showed much more potent effect than the unhydrolyzed one. In sharp contrast, those fractions of indigo stems had almost no effect. 3,5,4′-Trihydroxy-6,7-methylenedioxyflavone (TMF)-O-glycosides and tryptanthrin were detected exclusively in the extracts of the leaves. The isolated flavonol species were furthermore tested for their anti-inflammatory activities against the synthesis of NO and prostaglandin E2 in the cultured macrophage cells. More potent anti-inflammatory effects were recognized with different aglycones of flavonols than their flavonol O-glycosides. Although the inhibitory effects of TMF were less effective than those of tryptanthrin, the levels of flavonol O-glycosides with TMF were much more abundant than those of tryptanthrin in the leaves. Oral administration of the fraction containing flavonol O-glycosides with TMF into mice revealed the detection of free TMF in the blood circulation, indicating that the aglycone moiety can be cleaved by digestive enzymes and absorbed in the gut. Alternatively, the assay of hydrophilic oxygen radical absorbance capacity revealed that the isolated species of flavonol O-glycosides with TMF and their aglycone had appreciable antioxidant activities. Taken together, our findings suggest that the predominant flavonol O-glycosides with TMF as an aglycone could be promising natural agents for the application to herbal medicine, nutraceuticals, and food additives.

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Abbreviations

TMF:

3,5,4′-Trihydroxy-6,7-methylenedioxyflavone

NO:

Nitric oxide

LPS:

Lipopolysaccharide

PG:

Prostaglandin

Trolox:

6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid

HPLC:

High-performance liquid chromatography

AAPH:

2,2′-Azobis(2-amidinopropane)dihydrochloride

MEM:

Minimal essential medium

D-PBS (−):

Dulbecco’s phosphate-buffered saline without Ca2+ and Mg2+ ions

FBS:

Fetal bovine serum

UPLC:

Ultra-performance liquid chromatography

ESI-TOF/MSE :

Electrospray ionization time-of-flight mass spectrometryE

PDA:

Photodiode array

GAE:

Gallic acid equivalent

H-ORAC:

Hydrophilic oxygen radical absorbance capacity

TE:

Trolox equivalent

ANOVA:

Analysis of variance

DW:

Dry weight

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Acknowledgements

We are very grateful to Dr. Yuko Nakamura at Applied Biotechnology, Food Research Institute Local Independent Administrative Institution Tottori Institute Industrial Technology, Sakaiminato, Tottori for instructing us the methods for the assays of NO synthesis and H-ORAC.

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Authors and Affiliations

  1. Department of Research and Development, Kotobuki Seika Co., Ltd., 2028 Hatagasaki, Yonago, 683-0845, Japan

    Shota Tokuyama-Nakai, Hideto Kimura & Tomoe Ishihara

  2. The United Graduate School of Agricultural Sciences, Tottori University, 4-101 Koyama-Minami, Tottori, 680-8553, Japan

    Shota Tokuyama-Nakai, Mitsuo Jisaka & Kazushige Yokota

  3. Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu-Cho, Matsue, Shimane, 690-8504, Japan

    Mitsuo Jisaka & Kazushige Yokota

Authors
  1. Shota Tokuyama-Nakai
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  2. Hideto Kimura
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  3. Tomoe Ishihara
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Correspondence to Kazushige Yokota.

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Tokuyama-Nakai, S., Kimura, H., Ishihara, T. et al. In Vitro Anti-inflammatory and Antioxidant Activities of 3,5,4′-Trihydroxy-6,7-Methylenedioxyflavone-O-Glycosides and Their Aglycone from Leaves of Polygonum tinctorium Lour. Appl Biochem Biotechnol 184, 414–431 (2018). https://doi.org/10.1007/s12010-017-2555-8

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  • DOI: https://doi.org/10.1007/s12010-017-2555-8

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