Cytoprotective mechanism of heat shock protein 70 against hypoxia/reoxygenation injury.
The role of heat shock protein 70 (HSP70) in the cytoprotection against hypoxia/reoxygenation injury was examined. Adult rat cardiomyocytes were isolated, subjected to hyperthermia at 42 degrees C for 15 min (heat shock treatment), and then incubated at 37 degrees C for 3 to 24 h (HSP production process). Heat shock treatment increased HSP70 production (80-260% increase); the peak increase was seen after 9 h of HSP production process. Thereafter, the cells were subjected to 120-min hypoxia and 15-min reoxygenation. Heat shock treatment increased the survival of the cells subjected to hypoxia/reoxygenation (1.5-2.5-fold); the maximal cytoprotection was observed after 12 h of HSP production process. Heat shock treatment increased HSP70 content in the nucleus when cells were subjected to 12 h of HSP production process. To examine the role of HSP70 accumulation in the nuclear fraction, the activity of poly(ADP-ribose) synthetase (PARS), which functions in the nucleus and consumes high-energy phosphates excessively in the reoxygenated state, were measured in the cells with heat shock and 12 h of HSP production process. Heat shock treatment attenuated the hypoxia/reoxygenation-induced increase in the PARS activity (50% decrease). Treatment of the cells with 3-aminobenzamide, an inhibitor of PARS, exerted the effects similar to those of heat shock treatment. These results suggest that attenuation of the PARS activity in the nucleus may play an important role in the cytoprotective effect of HSP70 on hypoxia/reoxygenation injury.
Kawana
K
K
Department of Pharmacology, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Japan.
Miyamoto
Y
Y
Tanonaka
K
K
Han-no
Y
Y
Yoshida
H
H
Takahashi
M
M
Takeo
S
S
D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western; D015687:Cell Hypoxia; D002467:Cell Nucleus / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism; D019610:Cytoprotection; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005334:Fever; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology*; D006358:Hot Temperature; D008297:Male; D009206:Myocardium / Q000166:cytology*; D010710:Phosphates / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature; D013997:Time Factors
D001549:Benzamides; D004791:Enzyme Inhibitors; D018840:HSP70 Heat-Shock Proteins; D010710:Phosphates; C025160:3-aminobenzamide; D011065:Poly(ADP-ribose) Polymerases; D003402:Creatine Kinase