Protective effect of anisodamine against myocardial cell apoptosis through mitochondria impairment in cardiac arrest in pigs

Abstract

To investigate the protective effect of anisodamine on myocardial mitochondrial damage in cardiac arrest (CA) in pigs. Twenty-three male pigs were randomly divided into three groups, epinephrine group (n=9), anisodamine group (n=9) and control group (n=5). CA following ventricular fibrillation (VF) was induced by alternating current. The blood samples were collected before CA, 8 minutes after CA and instantly after recovery of spontaneous circulation (ROSC), and 30 minutes and 24 hours later. Hearts were obtained at 24 hours after ROSC. The changes in Cytochrome C (Cyt C) and caspase-3 in plasma and myocardium were analyzed by enzyme-linked immunosorbent assay (ELISA). The myocardial specimens were observed by transmission electron microscopy for ultrastructural changes, and apoptosis was assessed with Hoechst 33258 staining. The ROSC rate of the anisodamine group was elevated by 22.22% compared with the epinephrine group (77.78% vs. 55.56%, P>0.05). All animals with resumption of ROSC survived up to 24 hours. The plasma contents of Cyt C and caspase-3 in the epinephrine group and the anisodamine group gradually increased after ROSC, and were significantly higher than those in the control group. But the plasma Cyt C level in the anisodamine group was lower than that in the epinephrine group at 30 minutes and 24 hours after ROSC (48.68±19.50 nmol/L vs. 77.51±29.87 nmol/L, 48.98±20.26 nmol/L vs. 82.11±25.09 nmol/L, both P<0.05). There was no significant difference in protein contents of both Cyt C and caspase-3 in plasma and myocardium between two resuscitate groups. Both epinephrine and anisodamine could mitigate cardiac mitochondrial damage after CA, but the anisodamine showed better effect. The myocardium apoptosis ratio in the anisodamine group was lower than that of the epinephrine group [(0.15±0.04)% vs. (0.37±0.04)%, P<0.01]. By decreasing the protein content of Cyt C, and reducing the extent of damage to myocardial mitochondria, anisodamine can protect the myocardial ultrastructure, and restrain the mitochondria-induced cell apoptosis after resuscitation.