Amyloid β25-35 induced ROS-burst through NADPH oxidase is sensitive to iron chelation in microglial Bv2 cells.
Iron chelation therapy and inhibition of glial nicotinamide adenine dinucleotide phosphate (NADPH) oxidase can both represent possible routes for Alzheimer's disease modifying therapies. The metal hypothesis is largely focused on direct binding of metals to the N-terminal hydrophilic 1-16 domain peptides of Amyloid beta (Aβ) and how they jointly give rise to reactive oxygen species (ROS) production. The cytotoxic effects of Aβ through ROS and metals are mainly studied in neuronal cells using full-length Aβ1-40/42 peptides. Here we study cellularly-derived ROS during 2-60min in response to non-metal associated mid domain Aβ25-35 in microglial Bv2 cells by fluorescence based spectroscopy. We analyze if Aβ25-35 induce ROS production through NADPH oxidase and if the production is sensitive to iron chelation. NADPH oxidase inhibitor diphenyliodonium (DPI) is used to confirm the production of ROS through NADPH oxidase. We modulate cellular iron homeostasis by applying cell permeable iron chelators desferrioxamine (DFO) and deferiprone (DFP). NADPH oxidase subunit gp91-phox level was analyzed by Western blotting. Our results show that Aβ25-35 induces strong ROS production through NADPH oxidase in Bv2 microglial cells. Intracellular iron depletion resulted in restrained Aβ25-35 induced ROS.
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia. Electronic address: kristin2@tlu.ee.
Kristin
K
Part
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia; Laboratory of Molecular Genetics National Institute of Chemical Physics and Biophysics, Akadeemia tee 23, 12618 Tallinn, Estonia. Electronic address: kai.kunnis@kbfi.ee.
Kai
K
Künnis-Beres
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia. Electronic address: helenposka@gmail.com.
Helen
H
Poska
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia. Electronic address: tiit.land@tlu.ee.
Tiit
T
Land
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia. Electronic address: ruth.shimmo@tlu.ee.
Ruth
R
Shimmo
School of Natural Sciences and Health, Tallinn University, Narva Mnt 29, 10120 Tallinn, Estonia. Electronic address: sandra.fernaeus@tlu.ee.
Sandra
S
Zetterström Fernaeus
D016229:Amyloid beta-Peptides / Q000633:toxicity*; D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D007501:Iron / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000494:pharmacology*; D051379:Mice; D017628:Microglia / Q000187:drug effects / Q000378:metabolism*; D019255:NADPH Oxidases / Q000378:metabolism*; D010446:Peptide Fragments / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism*
D016229:Amyloid beta-Peptides; D007502:Iron Chelating Agents; D010446:Peptide Fragments; D017382:Reactive Oxygen Species; C074636:amyloid beta-protein (25-35); D007501:Iron; D019255:NADPH Oxidases
Aβ(25–35); Iron; Metal chelation; Microglia; NADPH oxidase; Reactive oxygen species