title: Fluctuating vs. continuous exposure to H₂O₂: the effects on mitochondrial membrane potential, intracellular calcium, and NF-κB in astroglia.
abstract: The effects of H2O2 are widely studied in cell cultures and other in vitro systems. However, such investigations are performed with the assumption that H2O2 concentration is constant, which may not properly reflect in vivo settings, particularly in redox-turbulent microenvironments such as mitochondria. Here we introduced and tested a novel concept of fluctuating oxidative stress. We treated C6 astroglial cells and primary astrocytes with H2O2, using three regimes of exposure - continuous, as well as fluctuating at low or high rate, and evaluated mitochondrial membrane potential and other parameters of mitochondrial activity - respiration, reducing capacity, and superoxide production, as well as intracellular ATP, intracellular calcium, and NF-κB activation. When compared to continuous exposure, fluctuating H2O2 induced a pronounced hyperpolarization in mitochondria, whereas the activity of electron transport chain appears not to be significantly affected. H2O2 provoked a decrease of ATP level and an increase of intracellular calcium concentration, independently of the regime of treatment. However, fluctuating H2O2 induced a specific pattern of large-amplitude fluctuations of calcium concentration. An impact on NF-κB activation was observed for high rate fluctuations, whereas continuous and low rate fluctuating oxidative stress did not provoke significant effects. Presented results outline the (patho)physiological relevance of redox fluctuations.
authors.affiliation: Center for Laser Microscopy, Faculty of Biology, University of Belgrade, Belgrade, Serbia.
authors.forename: Aleksandar
authors.initials: A
authors.lastname: Bajić
authors.forename: Mihajlo
authors.initials: M
authors.lastname: Spasić
authors.forename: Pavle R
authors.initials: PR
authors.lastname: Andjus
authors.forename: Danijela
authors.initials: D
authors.lastname: Savić
authors.forename: Ana
authors.initials: A
authors.lastname: Parabucki
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Nikolić-Kokić
authors.forename: Ivan
authors.initials: I
authors.lastname: Spasojević
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D042541:Intracellular Space / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D018384:Oxidative Stress; D061251:Primary Cell Culture; D021381:Protein Transport; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D016328:NF-kappa B; D013438:Sulfhydryl Compounds; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D002118:Calcium