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Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin

Fisetin, a dietary flavonoid, is reported to have cellular antioxidant activity with an unclear mechanism. In this study, we investigated the effect of fisetin on the nuclear factor, erythroid 2-like 2 (Nrf2) signaling pathway in HepG2 cells to explore the cellular antioxidant mechanism. Fisetin upr...

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Autores principales: Zhang, Huihui, Zheng, Wan, Feng, Xiangling, Yang, Fei, Qin, Hong, Wu, Shusong, Hou, De-Xing, Chen, Jihua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6413105/
https://www.ncbi.nlm.nih.gov/pubmed/30781396
http://dx.doi.org/10.3390/molecules24040708
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author Zhang, Huihui
Zheng, Wan
Feng, Xiangling
Yang, Fei
Qin, Hong
Wu, Shusong
Hou, De-Xing
Chen, Jihua
author_facet Zhang, Huihui
Zheng, Wan
Feng, Xiangling
Yang, Fei
Qin, Hong
Wu, Shusong
Hou, De-Xing
Chen, Jihua
author_sort Zhang, Huihui
collection PubMed
description Fisetin, a dietary flavonoid, is reported to have cellular antioxidant activity with an unclear mechanism. In this study, we investigated the effect of fisetin on the nuclear factor, erythroid 2-like 2 (Nrf2) signaling pathway in HepG2 cells to explore the cellular antioxidant mechanism. Fisetin upregulated the mRNA expression of heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), and NAD(P)H quinone oxidoreductase-1 (NQO1), and induced the protein of HO-1 but had no significant effect on the protein of GCLC, GCLM and NQO1. Moreover, nuclear accumulation of Nrf2 was clearly observed by immunofluorescence analysis and western blotting after fisetin treatment, and an enhanced luciferase activity of antioxidant response element (ARE)-regulated transactivation was obtained by dual-luciferase reporter gene assays. In addition, fisetin upregulated the protein level of Nrf2 and downregulated the protein level of Kelch-like ECH-associated protein 1 (Keap1). However, fisetin had no significant effect on Nrf2 mRNA expression. When protein synthesis was inhibited with cycloheximide (CHX), fisetin prolonged the half-life of Nrf2 from 15 min to 45 min. When blocking Nrf2 degradation with proteasome inhibitor MG132, ubiquitinated proteins were enhanced, and fisetin reduced ubiquitination of Nrf2. Taken together, fisetin translocated Nrf2 into the nucleus and upregulated the expression of downstream HO-1 gene by inhibiting the degradation of Nrf2 at the post-transcriptional level. These data provide the molecular mechanism to understand the cellular antioxidant activity of fisetin.
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spelling pubmed-64131052019-03-29 Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin Zhang, Huihui Zheng, Wan Feng, Xiangling Yang, Fei Qin, Hong Wu, Shusong Hou, De-Xing Chen, Jihua Molecules Article Fisetin, a dietary flavonoid, is reported to have cellular antioxidant activity with an unclear mechanism. In this study, we investigated the effect of fisetin on the nuclear factor, erythroid 2-like 2 (Nrf2) signaling pathway in HepG2 cells to explore the cellular antioxidant mechanism. Fisetin upregulated the mRNA expression of heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), and NAD(P)H quinone oxidoreductase-1 (NQO1), and induced the protein of HO-1 but had no significant effect on the protein of GCLC, GCLM and NQO1. Moreover, nuclear accumulation of Nrf2 was clearly observed by immunofluorescence analysis and western blotting after fisetin treatment, and an enhanced luciferase activity of antioxidant response element (ARE)-regulated transactivation was obtained by dual-luciferase reporter gene assays. In addition, fisetin upregulated the protein level of Nrf2 and downregulated the protein level of Kelch-like ECH-associated protein 1 (Keap1). However, fisetin had no significant effect on Nrf2 mRNA expression. When protein synthesis was inhibited with cycloheximide (CHX), fisetin prolonged the half-life of Nrf2 from 15 min to 45 min. When blocking Nrf2 degradation with proteasome inhibitor MG132, ubiquitinated proteins were enhanced, and fisetin reduced ubiquitination of Nrf2. Taken together, fisetin translocated Nrf2 into the nucleus and upregulated the expression of downstream HO-1 gene by inhibiting the degradation of Nrf2 at the post-transcriptional level. These data provide the molecular mechanism to understand the cellular antioxidant activity of fisetin. MDPI 2019-02-15 /pmc/articles/PMC6413105/ /pubmed/30781396 http://dx.doi.org/10.3390/molecules24040708 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Huihui
Zheng, Wan
Feng, Xiangling
Yang, Fei
Qin, Hong
Wu, Shusong
Hou, De-Xing
Chen, Jihua
Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title_full Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title_fullStr Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title_full_unstemmed Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title_short Nrf2–ARE Signaling Acts as Master Pathway for the Cellular Antioxidant Activity of Fisetin
title_sort nrf2–are signaling acts as master pathway for the cellular antioxidant activity of fisetin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6413105/
https://www.ncbi.nlm.nih.gov/pubmed/30781396
http://dx.doi.org/10.3390/molecules24040708
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