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Int. J. Mol. Sci. 2015, 16(9), 21410-21427; doi:10.3390/ijms160921410

The Effects of Cadmium at Low Environmental Concentrations on THP-1 Macrophage Apoptosis

1
Department of Hygiene and Epidemiology, Pomeranian Medical University, Szczecin 70-111, Poland
2
Department of Biochemistry and Medical Chemistry, Pomeranian Medical University, Szczecin 70-111, Poland
3
Department of Biochemistry and Human Nutrition, Pomeranian Medical University, Szczecin 71-460, Poland
4
Department of Physiology, Pomeranian Medical University, Szczecin 70-111, Poland
5
Department of Experimental and Clinical Pharmacology, Pomeranian Medical University, Szczecin 70-111, Poland
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Bing Yan
Received: 10 July 2015 / Revised: 13 August 2015 / Accepted: 19 August 2015 / Published: 7 September 2015
(This article belongs to the Section Molecular Toxicology)
View Full-Text   |   Download PDF [2157 KB, uploaded 7 September 2015]   |  

Abstract

Cadmium 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
Keywords: cadmium; apoptosis; ROS; mitochondrial membrane potential; Bax; Bcl-2; THP-1 macrophages cadmium; apoptosis; ROS; mitochondrial membrane potential; Bax; Bcl-2; THP-1 macrophages
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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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.

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