Proanthocyanidins Activate Nrf2/ARE Signaling Pathway in Intestinal Epithelial Cells by Inhibiting the Ubiquitinated Degradation of Nrf2

Nrf2 plays a key role in the antioxidant system, and many antioxidants can activate the Nrf2/ARE signaling pathway and alleviate oxidative stress. However, the underlying mechanisms of antioxidants, such as proanthocyanidin- (PC-) induced Nrf2 activation, remain poorly understood. In this study, PC...

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Published in:BioMed research international 2022-08, Vol.2022, p.1-10
Main Authors: Shuhua, Yang, Lingqi, Meng, Yunlong, Dang, He, Tang, Yao, Shi, Peng, Li
Format: Article
Language:English
Subjects:
Analysis
Antibodies
Antioxidants
Biodegradation
Cell growth
Decomposition (Chemistry)
Degradation
Epithelial cells
Epithelium
Fluorescent antibody technique
Gene expression
Genetic aspects
Identification and classification
Immunofluorescence
Immunoprecipitation
Intestine
Methods
NQO1 protein
NRF2 protein
Oxidative stress
Physiology
Polyphenols
Proanthocyanidins
Properties
Protein expression
Proteins
Signal transduction
Signaling
Stability
Western blotting
Western immunoblotting
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Peng, Li
description Nrf2 plays a key role in the antioxidant system, and many antioxidants can activate the Nrf2/ARE signaling pathway and alleviate oxidative stress. However, the underlying mechanisms of antioxidants, such as proanthocyanidin- (PC-) induced Nrf2 activation, remain poorly understood. In this study, PC was used on MODE-K cells at different concentrations (0, 1, 2.5, and 5 μg/mL) and different times (0, 3, 6, 12, and 24 h); then, immunoprecipitation, immunofluorescence, and Western blotting were performed to test Nrf2, Bach1, Keap1, HO-1, and NQO1 protein expressions in MODE-K cells. Results showed that PC increased Nrf2, HO-1, and NQO1 protein expressions, decreased Keap1 and Bach1 protein expressions, and enhanced ARE gene activity. PC also decreased the ubiquitinated degradation of the Nrf2 protein, increased Nrf2 protein stability, and increased Nrf2 protein expression by inhibiting Keap1-dependent Nrf2 protein degradation, promoted Nrf2 entry into the nucleus, competed with Bach1, and activated ARE elements, which in turn initiated the Nrf2/ARE signaling pathway. Thus, we conclude that PC activates the Nrf2/ARE signaling pathway in intestinal epithelial cells by inhibiting the ubiquitinated degradation of Nrf2, increasing Nrf2 protein stability and expression, and then regulating key antioxidant enzymes such as HO-1 and NQO1 to initiate cytoprotective effects.
doi_str_mv 10.1155/2022/8562795
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However, the underlying mechanisms of antioxidants, such as proanthocyanidin- (PC-) induced Nrf2 activation, remain poorly understood. In this study, PC was used on MODE-K cells at different concentrations (0, 1, 2.5, and 5 μg/mL) and different times (0, 3, 6, 12, and 24 h); then, immunoprecipitation, immunofluorescence, and Western blotting were performed to test Nrf2, Bach1, Keap1, HO-1, and NQO1 protein expressions in MODE-K cells. Results showed that PC increased Nrf2, HO-1, and NQO1 protein expressions, decreased Keap1 and Bach1 protein expressions, and enhanced ARE gene activity. PC also decreased the ubiquitinated degradation of the Nrf2 protein, increased Nrf2 protein stability, and increased Nrf2 protein expression by inhibiting Keap1-dependent Nrf2 protein degradation, promoted Nrf2 entry into the nucleus, competed with Bach1, and activated ARE elements, which in turn initiated the Nrf2/ARE signaling pathway. Thus, we conclude that PC activates the Nrf2/ARE signaling pathway in intestinal epithelial cells by inhibiting the ubiquitinated degradation of Nrf2, increasing Nrf2 protein stability and expression, and then regulating key antioxidant enzymes such as HO-1 and NQO1 to initiate cytoprotective effects.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2022/8562795</identifier><identifier>PMID: 36033575</identifier><language>eng</language><publisher>New York: Hindawi</publisher><subject>Analysis ; Antibodies ; Antioxidants ; Biodegradation ; Cell growth ; Decomposition (Chemistry) ; Degradation ; Epithelial cells ; Epithelium ; Fluorescent antibody technique ; Gene expression ; Genetic aspects ; Identification and classification ; Immunofluorescence ; Immunoprecipitation ; Intestine ; Methods ; NQO1 protein ; NRF2 protein ; Oxidative stress ; Physiology ; Polyphenols ; Proanthocyanidins ; Properties ; Protein expression ; Proteins ; Signal transduction ; Signaling ; Stability ; Western blotting ; Western immunoblotting</subject><ispartof>BioMed research international, 2022-08, Vol.2022, p.1-10</ispartof><rights>Copyright © 2022 Yang Shuhua et al.</rights><rights>COPYRIGHT 2022 John Wiley &amp; Sons, Inc.</rights><rights>Copyright © 2022 Yang Shuhua et al. 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However, the underlying mechanisms of antioxidants, such as proanthocyanidin- (PC-) induced Nrf2 activation, remain poorly understood. In this study, PC was used on MODE-K cells at different concentrations (0, 1, 2.5, and 5 μg/mL) and different times (0, 3, 6, 12, and 24 h); then, immunoprecipitation, immunofluorescence, and Western blotting were performed to test Nrf2, Bach1, Keap1, HO-1, and NQO1 protein expressions in MODE-K cells. Results showed that PC increased Nrf2, HO-1, and NQO1 protein expressions, decreased Keap1 and Bach1 protein expressions, and enhanced ARE gene activity. PC also decreased the ubiquitinated degradation of the Nrf2 protein, increased Nrf2 protein stability, and increased Nrf2 protein expression by inhibiting Keap1-dependent Nrf2 protein degradation, promoted Nrf2 entry into the nucleus, competed with Bach1, and activated ARE elements, which in turn initiated the Nrf2/ARE signaling pathway. 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source Wiley-Blackwell Open Access Collection; Publicly Available Content Database
subjects Analysis
Antibodies
Antioxidants
Biodegradation
Cell growth
Decomposition (Chemistry)
Degradation
Epithelial cells
Epithelium
Fluorescent antibody technique
Gene expression
Genetic aspects
Identification and classification
Immunofluorescence
Immunoprecipitation
Intestine
Methods
NQO1 protein
NRF2 protein
Oxidative stress
Physiology
Polyphenols
Proanthocyanidins
Properties
Protein expression
Proteins
Signal transduction
Signaling
Stability
Western blotting
Western immunoblotting
title Proanthocyanidins Activate Nrf2/ARE Signaling Pathway in Intestinal Epithelial Cells by Inhibiting the Ubiquitinated Degradation of Nrf2
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