Abstract
Lipid peroxidation products (LPPs) generated by oxidative stress (OS) have been recognized as biomarkers of numerous human disorders including atherosclerosis, neurodegenerative diseases, type 2 diabetes, chronic inflammation, aging, and cardiovascular diseases. To evaluate the effect of OS and related LPPs on cardiac system, rat primary cardiomyocytes treated with peroxynitrite donor SIN-1 were used as a cellular model of mild nitroxidative stress. Carbonylated molecules were visualized by fluorescent microscopy using cell permeable dye 7-(diethylamino)coumarin-3-carbohydrazide (CHH). Interestingly, increase in lipid droplets (LDs; visualized using Nile Red staining) as well as their enrichment in carbonylated LPPs was observed upon SIN-1 treatment of cardiomyocytes. Furthermore, using Nile Red and CHH co-staining heterogeneity in LDs carbonylation was demonstrated. Role of autophagy-lysosomal degradation pathway in removal of carbonylated LPPs from LDs was further evaluated using selective inhibitors including 3-methyladenine (inhibitor of an autophagosome formation), chloroquine (inhibitor of an autophagosome and lysosome fusion), orlistat (lipase inhibitor), E64d and Pepstatine A (proteases inhibitors). Significant role of autophagy-lysosomal degradation pathway in elimination of oxidized lipids incorporated in OS induced LDs was demonstrated.
Published Version
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