Heat shock pretreatment inhibits the release of Smac from mitochondria and decreases H2O2-induced cardiomyocyte apoptosis
Jiang, B.-m.; Xiao, W.-m.; Shi, Y.-z.
Hunan Yi Ke da Xue Xue Bao 28(6): 567-571
2003
ISSN/ISBN: 1000-5625 PMID: 15804062 Document Number: 554867
To explore the effect of heat shock pretreatment on H2O2-induced apoptosis of neonatal rat cardiomyocytes and the release of Smac from mitochondria. After heat shock pretreatment (42 degrees C for 1 h), neonatal cardiomyocytes were exposed to H2O2 (0.5 mmol/L) for 6, 12, 24, and 36 h, respectively. The apoptotic morphological changes and percentage of apoptotic nuclei were analyzed. Activities of caspase-3, 9 were assayed with caspase colorimetric assay kit and Western-blotting. Inducible heat shock proteins were detected by Western-blotting analysis. The release of Smac from mitochondria to cytoplasm was observed by Western-blotting and immunofluorescence. (1) H2O2 (0.5 mmol/L) resulted in a marked release of Smac from mitochondria to cytoplasm at 2 h after the treatment, the activation of caspase-9 and caspase-3 at 4 h after the treatment and specific morphological changes of apoptosis at 24 h after the treatment. (2) Heat shock pretreatment (42 degrees C, 1 h) could increase the expression of hsp90, hsp70 and betaB-crystallin, and inhibit the H2O2-induced release of Smac from mitochondria, the activity of caspase-9, caspase-3 and apoptosis of cardiomyocytes. (1) Mitochondrial signal transduction pathway is involved in the apoptosis of cardiomyocytes induced by H2O2; (2) Heat shock pretreatment can inhibit the release of Smac from mitochondria and the activities of caspase-9, 3 and protect cardiomyocytes against H2O2-induced apoptosis.