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The Effect of Black Seed (Nigella sativa ) Extract on FOXO3 Expression in HepG2 Cells
Author(s) -
Haas Michael J.,
OnsteadHaas Luisa,
Naem Emad,
Arnold Alexis,
Rohrbaugh Nathcelly,
Flowers Megan,
Mooradian Arshag D.
Publication year - 2014
Publication title -
phytotherapy research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.019
H-Index - 129
eISSN - 1099-1573
pISSN - 0951-418X
DOI - 10.1002/ptr.5069
Subject(s) - foxo3 , nigella sativa , phosphorylation , western blot , protein kinase b , ampk , microbiology and biotechnology , transcription factor , biology , signal transduction , kinase , protein kinase a , biochemistry , gene , traditional medicine , medicine
Black seed extracts are known to alter cellular metabolism through multiple signaling pathways. Since Forkhead box transcription factor 3 (FOXO3) has a significant role in regulating cellular metabolism, the effect of lipid extracts of black seed (Sativa nigella ) on FOXO3 levels and AKT and 5‐AMP activated protein kinase α (AMPKα) signaling was measured in HepG2 hepatoma cells. FOXO3 levels, phosphorylation, and nuclear exclusion were measured by Western blot, as were AKT and AMPK expression and activity using phosphorylation‐specific antibodies. Apolipoprotein A‐I expression, a black seed‐responsive gene, was measured by Western blot. Treatment with black seed extract increased FOXO3 phosphorylation and decreased its expression. In contrast to control cells where FOXO3 was located primarily in the nucleus, in black seed‐treated HepG2 cells, FOXO3 was localized primarily to the cytoplasm. These changes in FOXO3 phosphorylation, expression, and localization were accompanied by increased AKT activity. Black seed also decreased AMPKα activity but increased AMPKα expression. Lipid extracts from black seeds inhibit FOXO3 activity and thereby modulate the expression of FOXO3‐dependent genes. Copyright © 2013 John Wiley & Sons, Ltd.