The protective effect and mechanism of sivelestat on the heart after resuscitation through regulating β-catenin signaling pathway / 中华急诊医学杂志

ABSTRACT

Objective:To establish the pig model of cardiac arrest and resuscitation, and then investigate the protective role of sivelestat (SV) on the heart after resuscitation and its relation with β-catenin signaling pathway.Methods:Twenty-five healthy male white pigs were purchased. The animals were randomly divided into the Sham group ( n=6), cardiopulmonary resuscitation group (CPR, n=10), and CPR+SV group ( n=9). The experimental animal model was established by 9 min of cardiac arrest induced by the method of ventricular fibrillation and then 6 min of CPR in the CPR and CPR+SV groups. At 5 min after successful resuscitation, a dose of 10 mg/kg of SV was infused in a duration of 1h via the femoral vein with a micro-infusion pump in the CPR+SV group. Myocardial function evaluated by the values of stroke volume (SV) and global ejection fraction (GEF) was measured by PiCCO at baseline, and at 0.5, 1, 2, 4 h after resuscitation. The serum concentrations of cardiac injury biomarkers including cardiac troponin I (cTnI) and creatine kinase isoenzymes (CK-MB) were measured by ELISA using blood samples drawn from the femoral vein at baseline, and at 1, 2, 4, and 24 h after resuscitation. The animals were euthanized at 24 h after resuscitation, and then cardiac tissue samples were harvested to measure the protein expression levels of β-catenin, Cyclin D1, c-Myc, cleaved caspase-9, and cleaved caspase-3 by Western blot and the degree of cell apoptosis by TUNEL. Results:Prior to cardiac arrest, myocardial function and cardiac injury biomarkers were maintained at the same levels, and no differences were observed among the three groups (all P> 0.05). After resuscitation, myocardial dysfunction and cardiac injury were observed in the CPR and CPR+SV groups, in which the values of SV and GEF were significantly decreased and meanwhile the serum concentrations of cTnI and CKMB were significantly increased when compared with the Sham group (all P< 0.05). However, myocardial dysfunction and cardiac injury were significantly milder in the CPR+SV group, in which the value of SV at 4h post-resuscitation and the values of GEF starting 1h post-resuscitation were significantly increased, and the serum concentrations of cTnI and CKMB were significantly decreased at 4 and 24 h post-resuscitation when compared to the CPR group (all P< 0.05). Tissue measurements indicated that the change of β-catenin signaling pathway and the occurrence of cell apoptosis were observed in the heart at 24 h post-resuscitation in the CPR and CPR+SV groups, which were indicated by significant increases in the protein expression levels of β-catenin, Cyclin D1, c-Myc, cleaved caspase-9, and cleaved caspase-3, and marked elevation in the index of cell apoptosis when compared with the Sham groups (all P< 0.05). However, the expression levels of proteins mentioned above were significantly decreased in the heart at 24 h post-resuscitation and the index of cell apoptosis was significantly reduced in the CPR+SV group when compared to the CPR group (all P< 0.05). Conclusion:SV has the protective role in alleviating post-resuscitation myocardial dysfunction and cardiac injury, in which the protective mechanism is possibly related to the alleviation of cell apoptosis through the inhibition of β-catenin signaling pathway activation.