Application of Centrifugal Partition Chromatography for Bioactivity-Guided Purification of Antioxidant-Response-Element-Inducing Constituents from Atractylodis Rhizoma Alba.

Myeong Kim,Chul Kim,Ahmed Syed,Kevin Choe,Ji Kim,Young-Mi Kim

doi:10.3390/molecules23092274

Myeong Kim, Chul Kim + Show 4 more

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https://doi.org/10.3390/molecules23092274

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Abstract

Activity-guided separation of antioxidant response element (ARE)-inducing constituents from the rhizomes of Atractylodis Rhizoma Alba was performed by the combination of centrifugal partition chromatography (CPC) and an ARE luciferase reporter assay. From 3 g of the active n-hexane fraction, one polyacetylene, (6E,12E)-tetradeca-6,12-dien-8,10-diyne-1,3-diyl diacetate (47.3 mg), and two sesquiterpenes, atractylenolide I (40.9 mg), and selina-4(14),7(11)-dien-8-one (6.0 mg) were successfully isolated by CPC with n-hexane–ethyl acetate–methanol–water (8:2:8:2, v/v). The chemical structures of the isolated compounds were determined by 1H- and 13C-NMR and ESI-MS. Among the isolated compounds, (6E,12E)-tetradeca-6,12-diene-8,10-diyne-1,3-diol diacetate and selina-4(14),7(11)-dien-8-one increased ARE activity 32.9-fold and 16.6-fold, respectively, without significant cytotoxicity, when 5 µM sulforaphane enhanced ARE activity 27.1-fold. However, atractylenolide I did not increase ARE activity at 100 µM, and showed cytotoxicity at concentrations over 10 µM.

Highlights

Oxidative stress is involved in various pathological conditions including ageing, cancer, inflammation, and chronic diseases [1]
Our results suggest that (6E,12E)-tetradeca-6,12-diene-8,10-diyne-1,3-diol diacetate (2) and selina-4(14),7(11)-dien-8-one (3) might reduce oxidative stress by enhancing cellular antioxidant systems without cytotoxicity and compounds 2 and 3 were major active molecules for antioxidant response element (ARE) induction activity in n-hexane extract of A. macrocephala
Bioactivity-guided isolation for ARE-inducing compounds was developed using a combination of centrifugal partition chromatography (CPC) and an ARE luciferase reporter assay

Summary

Introduction

Oxidative stress is involved in various pathological conditions including ageing, cancer, inflammation, and chronic diseases [1]. Reactive oxygen species can be scavenged by the use of antioxidants including vitamins, carotenoids, and polyphenolic compounds [2]. Induction of antioxidant signaling pathway via upregulation of endogenous antioxidant/phase II detoxifying enzymes may be an alternative defensive approach. 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway leads to the induction of antioxidant/phase II detoxifying enzymes including heme oxygenase 1, NAD(P)H: quinine oxidoreductase 1, and glutathione S-transferase. There is growing interest in molecules that activate the Nrf2-ARE signaling pathway [3].

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