Abstract
Samples of environmental particulate matter contain environmentally persistent free radicals (EPFRs) capable of sustained generation of oxygen radicals. While exposure to EPFRs produces cardiac toxicity and oxidative stress in experimental animals, the underlying mechanisms are largely unknown. To determine whether EPFRs could directly damage cardiomyocytes, cultured mouse cardiomyocytes (HL-1) and primary rat adult left ventricular myocytes (ALVM) were incubated with an EPFR consisting of 1,2-dichlorobenzene chemisorbed to CuO-coated silica beads (DCB230). Treatment with DCB230 killed both HL-1 and ALVM in a dose- and time-dependent manner. The cytotoxic effects of DCB230 were significantly attenuated by treatment with α-tocopherol. One to 2 h after exposure to DCB230, there were significant reductions in mitochondrial membrane potential and significant increases in cleaved caspase-9, but no significant increases in DNA damage or cell death. After 8 h of treatment, there were significant increases in caspase-3, caspase-9, DNA damage and PARP cleavage associated with significant cell death. Together, these data indicate that DCB230 kills HL-1 myocytes by inducing oxidative stress that initiates apoptosis, with the intrinsic or mitochondrial pathway acting early in the apoptotic signaling process.
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Silva, R., West, J., Zhang, Y., Anenberg, S., Lamarque, J., Shindell, D., et al. (2013). Global premature mortality due to anthropogenic outdoor air pollution and the contribution of past climate change. Environmental Research Letters, 121, 2331–2378.
Pope, C. A, 3rd, & Dockery, D. W. (2006). Health effects of fine particulate air pollution: Lines that connect. Journal of the Air and Waste Management Association, 56, 709–742.
Brook, R. D., Rajagopalan, S., Pope, C. A., Brook, J. R., Bhatnagar, A., Diez-Roux, A. V., et al. (2010). Particulate matter air pollution and cardiovascular disease: An update to the scientific statement From the American Heart Association. Circulation, 121, 2331–2378.
Nemmar, A., Vanbilloen, H., Hoylaerts, M. F., Hoet, P. H., Verbruggen, A., & Nemery, B. (2001). Passage of intratracheally instilled ultrafine particles from the lung into the systemic circulation in hamster. American Journal of Respiratory and Critical Care Medicine, 164, 1665–1668.
Takenaka, S., Karg, E., Roth, C., Schulz, H., Ziesenis, A., Heinzmann, U., et al. (2001). Pulmonary and systemic distribution of inhaled ultrafine silver particles in rats. Environmental Health Perspectives, 109, 547–551.
Dellinger, B., Pryor, W. A., Cueto, R., Squadrito, G. L., Hedge, V., & Deutsch, W. A. (2001). Role of free radicals in the toxicity of airborne fine particulate matter. Chemical Research in Toxicology, 14, 1371–1377.
Lord, K., Moll, D., Lindsey, J. K., Mahne, S., Raman, G., Dugas, T., et al. (2011). Environmentally persistent free radicals decrease cardiac function before and after ischemia/reperfusion injury in vivo. Journal of Receptors and Signal Transduction, 31, 157–167.
Mahne, S., Chuang, G. C., Pankey, E., Kiruri, L., Kadowitz, P. J., Dellinger, B., & Varner, K. J. (2012). Environmentally persistent free radicals decrease cardiac function and increase pulmonary artery pressure. American Journal of Physiology Heart and Circulatory Physiology, 303, H1135–H1142.
Burn, B., & Varner, K. (2015). Environmentally persistent free radicals (EPFRs) compromise left ventricular function during ischemia/reperfusion injury. American Journal of Physiology-Heart and Circulatory Physiology, 308, H998–H1006.
Araujo, J. A., Barajas, B., Kleinman, M., Wang, X., Bennett, B. J., Gong, K. W., et al. (2008). Ambient particulate pollutants in the ultrafine range promote early atherosclerosis and systemic oxidative stress. Circulation Research, 102, 589–596.
Balakrishna, S., Saravia, J., Thevenot, P., Ahlert, T., Lominiki, S., Dellinger, B., & Cormier, S. (2011). Environmentally persistent free radicals induce airway hyperresponsiveness in neonatal rat lungs. Particle and Fibre Toxicology, 8, 1743-8977–1748-1711.
Thevenot, P. T., Saravia, J., Jin, N., Giaimo, J. D., Chustz, R. E., Mahne, S., et al. (2013). Radical-containing ultrafine particulate matter initiates epithelial-to-mesenchymal transitions in airway epithelial cells. American Journal of Respiratory Cell and Molecular Biology, 48, 188–197.
Kelley, M., Hebert, V., Thibeaux, T., Orchard, M., Hasan, F., Cormier, S., et al. (2013). Model combustion-generated particulate matter containing persistent free radicals redox cycle to produce reactive oxygen species. Chemical Research in Toxicology, 26, 1862–1871.
Balakrishna, S., Lomnicki, S., McAvey, K. M., Cole, R. B., Dellinger, B., & Cormier, S. A. (2009). Environmentally persistent free radicals amplify ultrafine particle mediated cellular oxidative stress and cytotoxicity. Particle and Fibre Toxicology, 6, 11.
Fahmy, B., Ding, L., You, D., Lomnicki, S., Dellinger, B., & Cormier, S. A. (2010). In vitro and in vivo assessment of pulmonary risk associated with exposure to combustion generated fine particles. Environmental Toxicology and Pharmacology, 29, 173–182.
Lomnicki, S., Truong, H., Vejerano, E., & Dellinger, B. (2008). Copper oxide-based model of persistent free radical formation on combustion-derived particulate matter. Environmental Science and Technology, 42, 4982–4988.
Claycomb, W. C., Lanson, N. A, Jr, Stallworth, B. S., Egeland, D. B., Delcarpio, J. B., Bahinski, A., & Izzo, N. J, Jr. (1998). HL-1 cells: A cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. Proceedings of the National Academy of Sciences USA, 95, 2979–2984.
Shenouda, S., Varner, K., Carvalho, F., & Lucchesi, P. (2009). Metabolites of MDMA induce oxidative stress and contractile dysfunction in adult rat left ventricular myocytes. Cardiovascular Toxicology, 9, 30–38.
Bergmeyer, H., & Bernt, E. (1963). Lactate dehydrogenase in methods. In H. Bergmeyer (Ed.), Enzymatic analysis (pp. 574–578). London: Academic Press.
Elmore, S. P., Nishimura, Y., Qian, T., Herman, B., & Lemasters, J. J. (2004). Discrimination of depolarized from polarized mitochondria by confocal fluorescence resonance energy transfer. Archives of Biochemistry and Biophysics, 422, 145–152.
Lemasters, J. J., & Ramshesh, V. K. (2007). Imaging of mitochondrial polarization and depolarization with cationic fluorophores. Methods in Cell Biology, 80, 283–295.
Kim, J.-B., Kim, C., Choi, E., Park, S., Park, H., Pak, H.-N., et al. (2012). Particulate air pollution induces arrhythmia via oxidative stress and calcium calmodulin kinase II activation. Toxicology and Applied Pharmacology, 259, 66–73.
Knuckles, T. L., & Dreher, K. L. (2007). Fine oil combustion particle bioavailable constituents induce molecular profiles of oxidative stress, altered function, and cellular injury in cardiomyocytes. Journal of Toxicology and Environmental Health, Part A, 70, 1824–1837.
Landar, A., Zmijewski, J. W., Dickinson, D. A., Le Goffe, C., Johnson, M. S., Milne, G. L., Zanoni, G., Vidari, G., Morrow, J. D., & Darley-Usmar, V. M. (2006). Interaction of electrophilic lipid oxidation products with mitochondria in endothelial cells and formation of reactive oxygen species. American Journal of Physiology - Heart and Circulatory Physiology, 290, H1777–H1787.
Li, N., Sioutas, C., Cho, A., Schmitz, D., Misra, C., Sempf, J., et al. (2003). Ultrafine particulate pollutants induce oxidative stress and mitochondrial damage. Environmental Health Perspectives, 111, 455–460.
Calderon-Garciduenas, L., Gambling, T. M., Acuna, H., Garcia, R., Osnaya, N., Monroy, S., et al. (2001). Canines as sentinel species for assessing chronic exposures to air pollutants: Part 2. Cardiac pathology. Toxicological Sciences, 61, 356–367.
Villarreal-Calderon, R., Dale, G., Delgado-Chavez, R., Torres-Jardon, R., Zhu, H., Herritt, L., et al. (2012). Intra-city differences in cardiac expression of inflammatory genes and inflammasomes in young urbanites: A pilot study. Journal of Toxicologic Pathology, 25, 163–173.
Kodavanti, U. P., Moyer, C. F., Ledbetter, A. D., Schladweiler, M. C., Costa, D. L., Hauser, R., et al. (2003). Inhaled environmental combustion particles cause myocardial injury in the Wistar Kyoto rat. Toxicological Sciences, 71, 237–245.
Acknowledgments
The authors would also like to thank Dr. Charles Nichols for this assistance with the confocal microscopy and Dr. Martin Ronis for his insights and editorial suggestions.
Funding
This work was supported by the National Institutes of Health P42ES013648 and P30GM106392 to K.J.V.
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Chuang, G.C., Xia, H., Mahne, S.E. et al. Environmentally Persistent Free Radicals Cause Apoptosis in HL-1 Cardiomyocytes. Cardiovasc Toxicol 17, 140–149 (2017). https://doi.org/10.1007/s12012-016-9367-x
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DOI: https://doi.org/10.1007/s12012-016-9367-x