Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury.
OBJECTIVE
To explore whether overexpression of the small heat shock protein HSP20 in rat cardiomyocytes protects against simulated ischemia/reperfusion (SI/R) injury.


METHODS
Recombinant adenovirus expressing HSP20 was used to infect rat H9c2 cardiomyocytes at high efficiency, as assessed by green fluorescent protein. H9c2 cells were subjected to SI/R stress; survival was estimated through assessment of lactate dehydrogenase and cell apoptosis through caspase-3 activity.


RESULTS
Overexpression of HSP20 decreased lactate dehydrogenase release by 21.5% and caspase-3 activity by 58.8%. Pretreatment with the protein kinase C inhibitor Ro-31-8220 (0.1 micromol/L) for 30 min before SI/R canceled the protective effect of HSP20. The selective mitochondrial K+ATP channel inhibitor 5-hydroxydecanoate (100 micromol/L) had a similar effect. However, the non-selective K+ATP channel inhibitor glibenclamide (100 micromol/L) had no significant effect.


CONCLUSIONS
These data indicate that the protective effect of HSP20 in vitro is primarily due to reduced necrotic and apoptotic death of cardiomyocytes, possibly via the protein kinase C/mitochondrial K+ATP pathway.
Zhu
Yan-hui
YH
Department of Physiology and Pathophysiology, Basic Medical College, Key Laboratory of Molecular Cardiovascular Biology, Ministry of Education, Peking University, Beijing 100083, China.
Wang
Xian
X
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D003652:Decanoic Acids / Q000494:pharmacology; D050886:HSP20 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006880:Hydroxy Acids / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D017202:Myocardial Ischemia / Q000150:complications; D015428:Myocardial Reperfusion Injury / Q000209:etiology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D026902:Potassium Channel Blockers / Q000494:pharmacology; D015221:Potassium Channels / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D003652:Decanoic Acids; D050886:HSP20 Heat-Shock Proteins; D006880:Hydroxy Acids; D007211:Indoles; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D011994:Recombinant Proteins; C117780:mitochondrial K(ATP) channel; C052853:5-hydroxydecanoic acid; D007770:L-Lactate Dehydrogenase; D011493:Protein Kinase C; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; C064758:Ro 31-8220