Insulin treatment
In the preresuscitation insulin treated group, insulin 200 μU/g body weight in 1 ml saline was injected intramuscularly before resuscitation. The postresuscitation insulin group (HS-R-I), insulin was injected after resuscitation period of 60 min [Figure 1].
Measurement of tumor necrosis factor-alpha
Blood (0.5 ml) was collected from the left carotid artery cannula at the end of the experimental period harvesting the heart, and centrifuged at 2500 g for 10 min and plasma was stored at −80°C until analysis for TNF-a measurement. Serum samples were analyzed by ELISA (R&D systems).
Statistical analysis
Data were presented as means ± standard deviation Data were analyzed with one-way ANOVA. The values of P < 0.05 were considered significant. The Student's t-test was used to compare mean values between the two experimental groups.