title: Withaferin A induces heme oxygenase (HO-1) expression in endothelial cells via activation of the Keap1/Nrf2 pathway.
abstract: Withaferin A (WA), a natural phytochemical derived from the plant Withania somnifera, is a well-studied bioactive compound exerting a broad spectrum of health promoting effects. To gain better insight in the potential therapeutic capacity of WA, we evaluated the transcriptional effects of WA on primary human umbilical vein endothelial cells (HUVECs) and an endothelial cell line (EA.hy926). RNA microarray analysis of WA treated HUVEC cells demonstrated increased expression of the antioxidant gene heme oxygenase (HO-1). Transcriptional regulation of this gene is strongly dependent on the transcription factor NF-E2-related factor 2 (Nrf2), which senses chemical changes in the cell and coordinates transcriptional responses to maintain chemical homeostasis via expression of antioxidant genes and cytoprotective Phase II detoxifying enzymes. Under normal conditions, Nrf2 is kept in the cytoplasm by Kelch-like ECH-associated protein 1 (Keap1), an adaptor protein controlling the half-life of Nrf2 via constant proteasomal degradation. In this study we demonstrate that WA time- and concentration-dependently induces HO-1 expression in endothelial cells via upregulation and increased nuclear translocation of Nrf2. According to the crucial negative regulatory role of Keap1 in Nrf2 expression levels, a direct interaction of WA with Keap1 could be demonstrated. In vitro and in silico evaluations suggest that specific cysteine residues in Keap1 might be involved in the interaction with WA.
authors.affiliation: Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST), Department of Biochemistry and Microbiology, Ghent University, Proeftuinstraat 86, Gent, Belgium. Electronic address: karen.heyninck@ugent.be.
authors.forename: Karen
authors.initials: K
authors.lastname: Heyninck
authors.affiliation: Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST), Department of Biochemistry and Microbiology, Ghent University, Proeftuinstraat 86, Gent, Belgium.
authors.forename: Linde
authors.initials: L
authors.lastname: Sabbe
authors.affiliation: Laboratory of Protein Science, Proteomics and Epigenetic Signaling (PPES), Department of Biomedical Sciences, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Chandra Sekhar
authors.initials: CS
authors.lastname: Chirumamilla
authors.affiliation: Laboratory of Protein Science, Proteomics and Epigenetic Signaling (PPES), Department of Biomedical Sciences, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Szarc Vel Szic
authors.affiliation: Laboratory of Medicinal Chemistry, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vander Veken
authors.affiliation: Department of Pharmacology and Toxicology, Maastricht University, Universiteitssingel 50, 6229ER Maastricht, The Netherlands.
authors.forename: Kristien J A
authors.initials: KJA
authors.lastname: Lemmens
authors.affiliation: School of Pharmacy, Pharmaceutical Chemistry, University of Eastern Finland, 70211 Kuopio, Finland.
authors.forename: Maija
authors.initials: M
authors.lastname: Lahtela-Kakkonen
authors.affiliation: Biomedical Informatics Research Center Antwerp (Biomina), University of Antwerp & University Hospital Antwerp, Campus Middelheim, Middelheimlaan 1, Antwerpen, Belgium; Advanced Database Research and Modelling (ADReM), Department of Mathematics & Computer Sciences, University of Antwerp (UA), Campus Middelheim, Middelheimlaan 1, Antwerpen, Belgium.
authors.forename: Stefan
authors.initials: S
authors.lastname: Naulaerts
authors.affiliation: Center of Medical Genetics, Department of Biomedical Sciences, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, Wilrijk, Belgium; Center for Oncological Research (CORE) Antwerp, Laboratory of Cancer Research and Clinical Oncology, University of Antwerp, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Ken
authors.initials: K
authors.lastname: Op de Beeck
authors.affiliation: Biomedical Informatics Research Center Antwerp (Biomina), University of Antwerp & University Hospital Antwerp, Campus Middelheim, Middelheimlaan 1, Antwerpen, Belgium; Advanced Database Research and Modelling (ADReM), Department of Mathematics & Computer Sciences, University of Antwerp (UA), Campus Middelheim, Middelheimlaan 1, Antwerpen, Belgium.
authors.forename: Kris
authors.initials: K
authors.lastname: Laukens
authors.affiliation: Center for Oncological Research (CORE) Antwerp, Laboratory of Cancer Research and Clinical Oncology, University of Antwerp, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Guy
authors.initials: G
authors.lastname: Van Camp
authors.affiliation: Department of Pharmacology and Toxicology, Maastricht University, Universiteitssingel 50, 6229ER Maastricht, The Netherlands.
authors.forename: Antje R
authors.initials: AR
authors.lastname: Weseler
authors.affiliation: Department of Pharmacology and Toxicology, Maastricht University, Universiteitssingel 50, 6229ER Maastricht, The Netherlands.
authors.forename: Aalt
authors.initials: A
authors.lastname: Bast
authors.affiliation: Department of Pharmacology and Toxicology, Maastricht University, Universiteitssingel 50, 6229ER Maastricht, The Netherlands.
authors.forename: Guido R M M
authors.initials: GRMM
authors.lastname: Haenen
authors.affiliation: Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST), Department of Biochemistry and Microbiology, Ghent University, Proeftuinstraat 86, Gent, Belgium.
authors.forename: Guy
authors.initials: G
authors.lastname: Haegeman
authors.affiliation: Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST), Department of Biochemistry and Microbiology, Ghent University, Proeftuinstraat 86, Gent, Belgium; Laboratory of Protein Science, Proteomics and Epigenetic Signaling (PPES), Department of Biomedical Sciences, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, Wilrijk, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Vanden Berghe
mesh_terms: D000072283:A549 Cells; D002470:Cell Survival / Q000187:drug effects; D018360:Crystallography, X-Ray; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D057809:HEK293 Cells; D051547:Heme Oxygenase-1 / Q000235:genetics* / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D050217:Metabolic Detoxication, Phase II / Q000235:genetics; D062105:Molecular Docking Simulation; D051267:NF-E2-Related Factor 2 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011485:Protein Binding; D000072417:Protein Domains; D017433:Protein Structure, Secondary; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic; D054358:Withanolides / Q000737:chemistry / Q000494:pharmacology*
chemical_list: C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D034741:RNA, Small Interfering; D054358:Withanolides; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1; C009684:withaferin A
keywords: Antioxidant; Endothelial cell; Molecular pharmacology; Phytochemical; Signal transduction; Transcription regulation