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. 2014 Aug 29;19(3):500–516. doi: 10.2478/s11658-014-0209-1

The effect of resveratrol and its methylthio-derivatives on the Nrf2-ARE pathway in mouse epidermis and HaCaT keratinocytes

Violetta Krajka-Kuźniak 1, Hanna Szaefer 1, Tomasz Stefański 2, Stanisław Sobiak 2, Michał Cichocki 1, Wanda Baer-Dubowska 1,✉
PMCID: PMC6276002  PMID: 25169438

Abstract

Resveratrol is the most extensively studied stilbene derivative. We previously showed that methylthiostilbenes were more effective inhibitors of CYP1A1 and 1B1 activity than resveratrol. In this study, we investigated whether resveratrol and its methylthio-substituted derivatives, i.e. 3-M-4′-MTS (S2), 3,5-DM-4′-MTS (S5) and 3,4,5-TM-4′-MTS (S7) could activate Nrf2 signaling in the mouse epidermis and in human keratinocytes. Western blot analysis showed translocation of Nrf2 from the cytosol to the nucleus in both models. All of the tested stilbenes increased GST activity, but resveratrol was the most effective inducer. Moreover, only resveratrol increased the protein level of GSTP in the mouse epidermis. GSTM was enhanced in HaCaT cells after the treatment with derivatives S2 and S5. The same effect was observed for GSTP in the case of compound S2. Resveratrol and its derivatives reduced the NQO2 protein level in HaCaT cells. Thus, it is possible that increased expression of GSTP or GSTM and GST activity was linked with NQO2 inhibition in these cells. The results of this study indicate that resveratrol and its methylthioderivatives activate Nrf2 not only in the mouse epidermis, but also in human keratinocytes. Upregulating GST isozymes might be particularly important for deactivating chemical carcinogens, such as PAH.

Keywords: Nrf2, GST, NQO, Resveratrol, Methylthiostilbenes, HaCaT cells, Mouse epidermis

Full Text

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Abbreviations used

AhR

aryl hydrocarbon receptor

ARE

antioxidant response element

CDNB

1-chloro-2,4-dinitrobenzene

CYPs

cytochromes P450

DMEM

Dulbecco’s modified Eagle’s medium

DMF

dimethylformamide

DMSO

dimethyl sulfoxide

DTT

dithiothreitol

FBS

fetal bovine serum

GSH

glutathione

GST

glutathione-S-transferase

GSTA

glutathione-S-transferase A

GSTM

glutathione-Stransferase M

GSTP

glutathione-S-transferase P

HaCaT

spontaneously immortalized human keratinocyte cell line

Keap1

Kelch-like ECH-associated protein 1

MTT

3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide

Nrf2

nuclear factor erythroid 2-related factor 2

NQO

NAD(P)H: quinone oxidoreductase

PAHs

polycyclic aromatic hydrocarbons

Res

resveratrol

S2

3-methoxy-4′-methylthio-transstilbene (3-M-4′-MTS)

S5

3,5-dimethoxy-4′-methylthio-trans-stilbene (3,5-DM-4′-MTS)

S7

3,4,5-trimethoxy-4′-methylthio-trans-stilbene (3,4,5-TM-4′-MTS)

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