The protective effect and mechanism of sivelestat on the heart after resuscitation through regulatingβ-catenin signaling pathway
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 Dl,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 1 h 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.
万方数据
维普
