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Activation of Nrf2 plays a role in the S‐equol‐mediated attenuation of oxidative stress‐induced endothelial cell injury (1146.3)
Author(s) -
Zhang Ting,
Mi Mantian,
Liang Xinyu,
Shi Linying
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1146.3
Subject(s) - equol , oxidative stress , chemistry , downregulation and upregulation , viability assay , western blot , umbilical vein , messenger rna , microbiology and biotechnology , cell , andrology , biochemistry , endocrinology , biology , medicine , in vitro , gene , daidzein , genistein
Objective To study the effect of S‐equol on the activation of Nrf2/ARE signaling pathway in human umbilical vein endothelial cells (HUVECs). Method HUVECs were used in this study. Dual luciferase reporter gene assay was used to determine the effect of S‐equol on Nrf2‐ARE‐Luc activity. Total and nuclear protein were extracted, western blot assay was used to determine the effect of S‐equol on Nrf2 and its downstream antioxidant gene HO‐1 protein expression. Real‐time PCR assay was used to detect the effect of S‐equol on Nrf2 and HO‐1 mRNA levels. Cells were pretreated with Znpp, the specific antagonist of HO‐1 and Nrf2‐siRNA, CCK‐8 assay was used to determine the effect of S‐equol on cell viability under oxidative stress. Results S‐equol (1 nM, 10 nM and 100 nM) dose‐dependently increased the activity of Nrf2‐ARE‐Luc. S‐equol (100 nM) up‐regulated Nrf2 and HO‐1 protein levels, which in a dose and time‐dependent in HO‐1 but no in Nrf2. S‐equol treatment induced Nrf2 nuclear translocation. S‐equol almost had no efffect on Nrf2 mRNA level, but a dose‐dependent upregulation of HO‐1 mRNA levels after treatment with S‐equol. Pretreatment of Znpp, the specific blocker of HO‐1 and Nrf2‐siRNA attenuated the protective effect of S‐equol under oxidative stress in HUVECs. Conclusion Physiological concentration (nM) of S‐equol activated Nrf2/ARE signaling pathway, the main antioxidant mechanism, in HUVECs, which plays a role in attenuation of S‐equol in oxidative stress‐induced cell injury. Grant Funding Source : Supported by The National Natural Science Foundation of China (81102129)