Antioxidants Inhibit Nuclear Export of Telomerase Reverse Transcriptase and Delay Replicative Senescence of Endothelial Cells

• Published in: Circulation research 2004-04, Vol.94 (6), p.768-775
• Main Authors: Haendeler, Judith, Hoffmann, Jörg, Diehl, J Florian, Vasa, Mariuca, Spyridopoulos, Ioakim, Zeiher, Andreas M, Dimmeler, Stefanie
• Format: Article
• Language: English
• Subjects: Acetylcysteine - pharmacology; Antioxidants - pharmacology; Atorvastatin Calcium; Biological and medical sciences; Cell Nucleus - metabolism; Cells, Cultured - cytology; Cells, Cultured - drug effects; Cells, Cultured - metabolism; Cellular Senescence - drug effects; Depression, Chemical; DNA Replication - drug effects; DNA, Mitochondrial - analysis; DNA-Binding Proteins; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Enzyme Inhibitors - pharmacology; Fundamental and applied biological sciences. Psychology; Heptanoic Acids - pharmacology; Humans; Hydrogen Peroxide - pharmacology; Oxidation-Reduction; Oxidative Stress - drug effects; Protein Transport - drug effects; Pyrimidines - pharmacology; Pyrroles - pharmacology; Reactive Oxygen Species - metabolism; src-Family Kinases - antagonists & inhibitors; src-Family Kinases - physiology; Telomerase - metabolism; Vertebrates: cardiovascular system
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/01.RES.0000121104.05977.F3

ABSTRACT—Aging is associated with a rise in intracellular reactive oxygen species (ROS) and a loss of telomerase reverse transcriptase activity. Incubation with H2O2 induced the nuclear export of telomerase reverse transcriptase (TERT) into the cytosol in a Src-family kinase–dependent manner. Therefore, we investigated the hypothesis that age-related increase in reactive oxygen species (ROS) may induce the nuclear export of TERT and contribute to endothelial cell senescence. Continuous cultivation of endothelial cells resulted in an increased endogenous formation of ROS starting after 29 population doublings (PDL). This increase was accompanied by mitochondrial DNA damage and preceded the onset of replicative senescence at PDL 37. Along with the enhanced formation of ROS, we detected an export of nuclear TERT protein from the nucleus into the cytoplasm and an activation of the Src-kinase. Moreover, the induction of premature senescence by low concentrations of H2O2 was completely blocked with the Src-family kinase inhibitor PP2, suggesting a crucial role for Src-family kinases in the induction of endothelial cell aging. Incubation with the antioxidant N-acetylcysteine, from PDL 26, reduced the intracellular ROS formation and prevented mitochondrial DNA damage. Likewise, nuclear export of TERT protein, loss in the overall TERT activity, and the onset of replicative senescence were delayed by incubation with N-acetylcysteine. Low doses of the statin, atorvastatin (0.1 μmol/L), had also effects similar to those of N-acetylcysteine. We conclude that both antioxidants and statins can delay the onset of replicative senescence by counteracting the increased ROS production linked to aging of endothelial cells.