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Abstract

Effects of heat shock protein 70 (Hsp70) translocated to nuclear fraction on hypoxia/reoxygenation injury was examined by using adult cardiomyocytes isolated from rats. Cardiomyocytes were exposed to heat shock at 42 degrees C for 15 min (HS group), and then incubated at 37 degrees C for 6-24 h. Hsp70 production increased and the protein translocated from cytosol to nucleus. The maximum level of Hsp70 in the nuclear fraction was observed 12 h after HS. When cardiomyocytes without exposure to HS (nHS group) were subjected to 120 min hypoxia/15 min reoxygenation (Hypo/Reoxy), post-hypoxic cell viability was approximately 25% of the pre-hypoxic value. A rise in poly(ADP-ribose) synthetase (PARS) activity in the nuclear fraction was observed in nHS group, associated with an increase in polyADP-ribosylated protein. In contrast, post-hypoxic cell viability of HS group was approximately 60% of the pre-hypoxic value. Hypo/reoxy-induced rise in PARS activity and increase in polyADP-ribosylated protein were attenuated in HS group. To confirm the relationship between an increase in cell viability after Hypo/Reoxy and attenuation of PARS activation, cardiomyocytes without exposure to HS were subjected to Hypo/Reoxy in the presence of 1 mM 3-aminobenzamide, an inhibitor of PARS. Treatment of cells with 3-aminobenzamide attenuated Hypo/ Reoxy-induced decrease in cell viability. These results suggest that Hsp70 translocated into nucleus after HS may attenuate PARS activation during Hypo/Reoxy, leading to the cytoprotection of cardiomyocytes against Hypo/Reoxy injury.