Different Role of Antioxidants in Endotoxin-Induced Late Myocardial Protection

Abstract

Previous studies have proposed that endogenous antioxidants play a protective role against cardiac ischemia-reperfusion injury in endotoxin pretreatment. However, the mechanism underlying this effect remains elusive. We therefore evaluated the role of endogenous antioxidants in delayed myocardial protection after different doses of endotoxin administration using cultured rat neonatal cardiomyocytes. Myocytes were treated with normal saline (control) or lipopolysaccharide (Escherichia coli,serotype O111) at doses of 40 and 80 μg/ml (ET40 and ET80). Also, antisense oligodeoxyribonucleotide (1.5 μmol/L) to manganese superoxide dismutase (Mn-SOD) and 3-amino-1,2,4-triazole (25 mg/ml) were added along with a 40 or 80 μg/ml endotoxin pretreatment in the IET40 and IET80 groups. Twenty-four hours later, Cells were subjected to hypoxia (pO2< 1 kPa, 3 h) and reoxygenation (pO2: 19 kPa, 1 h). Compared with controls, cell viability enhanced significantly (65.3 ± 5.9, 63.8 ± 4.6, and 69.7 ± 5.2% vs 47.2 ± 4.3%,P< 0.05) and creatine kinase release decreased (7.34 ± 1.76, 7.11 ± 1.49, and 6.27 ± 1.24 U/mg protein vs 11.23 ± 2.49 U/mg protein,P< 0.05) in ET40, IET40, and ET80 groups following reoxygenation. No statistically significant difference was found between the control and the IET80 groups. Furthermore, the levels of Mn-SOD (1.12 ± 0.31 vs 0.75 ± 0.15 U/mg · protein,P< 0.05) and catalase activity (1265 ± 109 vs 996 ± 85 U/mg · protein,P< 0.05) were higher only in the ET80 group. The results suggest that at a dose of 40 μg/ml, cells were protected by mechanisms other than the augmentation of endogenous antioxidant activity which were more evident at a dose of 80 μg/ml. It seems that different doses of endotoxin pretreatment may induce delayed myocardial protection through various mechanisms.