The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis
AbstractCadmium at environmental concentrations is a risk factor for many diseases, including cardiovascular and neurodegenerative diseases, in which macrophages play an important role. The aim of this study was to evaluate the effects of cadmium at low environmental (nanomolar) concentrations on apoptotic processes in THP-1(acute monocytic leukemia cells line)-derived macrophages, with special focus on mitochondrial events involved. Macrophages were incubated with various cadmium chloride (CdCl2) solutions for 48 h at final concentrations of 5 nM, 20 nM, 200 nM and 2 µM CdCl2. Cell viability was measured using flow cytometry. Flow cytometric measurement (annexin V/FITC (annexin V/fluorescein isothiocyanate) and PI (propidium iodide) double staining) was used to quantify the extent of apoptosis. Fluorescence and confocal microscopy were used for imaging of apoptosis process. Changes in mitochondrial membrane potential were monitored using cytofluorimetry after cell staining with JC-1(5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyane iodide) probe. Mitochondrial ROS (reactive oxygen species) levels were measured cytofluorimetrically after incubation of cells with mitochondrial superoxide indicator (MitoSOX) red fluorescent marker. The mRNA expression of Bcl-2 and Bax was analysed with qRT-PCR. Our study demonstrates that cadmium, even at low environmental concentrations, exerts mitochondrial toxicity in THP-1 macrophages. Forty-eight-hour exposure to very low concentrations reduces cell viability and results in cell death by apoptosis and necrosis. The decrease in mitochondrial membrane potential, increased ROS production, increased Bax and decreased Bcl-2 mRNA expression are mitochondrial events involved in cadmium-induced apoptosis. View Full-Text
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Olszowski, T.; Baranowska-Bosiacka, I.; Gutowska, I.; Piotrowska, K.; Mierzejewska, K.; Korbecki, J.; Kurzawski, M.; Tarnowski, M.; Chlubek, D. The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis. Int. J. Mol. Sci. 2015, 16, 21410-21427.
Olszowski T, Baranowska-Bosiacka I, Gutowska I, Piotrowska K, Mierzejewska K, Korbecki J, Kurzawski M, Tarnowski M, Chlubek D. The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis. International Journal of Molecular Sciences. 2015; 16(9):21410-21427.Chicago/Turabian Style
Olszowski, Tomasz; Baranowska-Bosiacka, Irena; Gutowska, Izabela; Piotrowska, Katarzyna; Mierzejewska, Katarzyna; Korbecki, Jan; Kurzawski, Mateusz; Tarnowski, Maciej; Chlubek, Dariusz. 2015. "The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis." Int. J. Mol. Sci. 16, no. 9: 21410-21427.