Expression of heme oxygenase-1 due to intracellular reactive oxygen species induced by ultrasound.
The present study was undertaken to elucidate the mechanism by which ultrasound induces the expression of heme oxygenase-1 (HO-1). When human lymphoma U937 cells were exposed to a 1 MHz continuous wave for 1 min, HO-1 expression examined by real-time quantitative polymerase chain reaction and immunoblotting was observed at intensities above the cavitational threshold. No induction of HO-1 expression was observed in the cells exposed for 1 min to 42 degrees C, a temperature higher than that during sonication. When a potent antioxidant, N-acetyl-l-cysteine, was added to the culture medium before or after sonication, the induction was attenuated, indicating that reactive oxygen species (ROS) are involved. However, the addition of catalase did not affect the induction, and no HO-1 was observed on the addition of pre-sonicated medium, suggesting that hydrogen peroxide due to the recombination of hydroxyl radicals generated extracellularly was not involved. The addition of free radical scavengers, glutathion-monoethyl ester, dimethyl sulfoxide and D(-)-mannitol, suppressed the induction. A decrease in mitochondrial membrane potential and the generation of superoxide were also observed in the sonicated cells, suggesting that mitochondria were the source of intracellularly generated ROS. These results indicate that superoxide secondarily generated from damaged mitochondria, not hydroxyl radicals generated in medium directly by sonication, give rise to intracellular oxidative stress inducing HO-1 expression.
Kagiya
Go
G
Medical Division, The Wakasa Wan Energy Research Center, 64-52-1 Nagatani, Tsuruga, Fukui 914-0192, Japan.
Ogawa
Ryohei
R
Tabuchi
Yoshiaki
Y
Feril
Loreto B
LB
Nozaki
Tetsuo
T
Fukuda
Shigekazu
S
Yamamoto
Kazutaka
K
Kondo
Takashi
T
D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D005978:Glutathione / Q000737:chemistry; D051547:Heme Oxygenase-1 / Q000096:biosynthesis*; D006801:Humans; D008564:Membrane Potentials; D018384:Oxidative Stress; D017382:Reactive Oxygen Species; D013010:Sonication; D013696:Temperature; D020298:U937 Cells; D014465:Ultrasonics
D000975:Antioxidants; D005609:Free Radicals; D017382:Reactive Oxygen Species; D051547:Heme Oxygenase-1; D005978:Glutathione; D000111:Acetylcysteine