Effects Of Anisodamine On Myocardial Ultrastructure Damage And Intercellular Signal Transmittion In Cardiac Arrest Swine

Objective Through the research on the oxygen radicals, mitochondria-induced andendoplasmic reticulum-induced cell apoptosis, observe the protection ofanisodamine on myocardial ultrastructure damage induced by ischemia reperfusioninjury (IRI) in post cardiac arrest(CA) resuscitation swine. And study the effect ofanisodamine on the myocardium intercellular signal transmittion.Methods1. After9min of electrically-induced CA, swine receivedcardiopulmonary resuscitation(CPR), and were randomly divided into groups withanisodamine administration (Ani+CA/R group, n=9)) and without anisodamineadministration (CA/R group, n=9). Restoration of spontaneous circulation(ROSC)rates and CPR time were calculated in each resuscitation group.2. The blood samplewere took at basic time, CA8min, ROSC, ROSC30min, ROSC24h, and heartswere harvested at24h after ROSC. The specimens were observed to assess theprotein changes of Cytochrome C and Caspase-3, and the enzyme activity ofsuperoxide dismutase(SOD), the content of malondialdehyde(MDA) were analyzedby chemical approach.3. The myocardial specimens were observed by transmissionelectron microscopy for ultrastructures, stained with Hoechst33258to assessapoptosis, by RT-PCR for mRNA level and western blotting for protein expressionof caspase-12, JNK, and calpain.4. The retrieved tissues were analyzed byimmunofluorescence for Cx43expression and distribution, by RT-PCR for themRNA level of Cx43and Calpain, and by western blotting to determine the levels ofp-Cx43, Cx43, and Calpain.Results1. The ventricular fibrillation was successful approached for all animals.The ROSC rate of Ani+CA/R group are higher than CA/R group,and a shorter CPRtime than CA/R group, with no statistic difference between two CPR group.2.① The myocardium SOD activity of Ani+CA/R group was higher than CA/R group（p<0.01）.②The MDA level of Ani+CA/R group was lower than CA/R group inROSC30min, ROSC24h(p<0.05）.③The level of blood Cyto C at ROSC30minand ROSC24h of Ani+CA/R group were lower than CA/R group（p<0.05）.④Thedifference between two CPR groups both in blood and myocardium Caspase-3levelhave no statistic meaning（p>0.05）.3. The CA/R group showed a more irregulararrangement of cardiomyocytes, seriously damaged sarcoplasmic reticula andmitochondria. Anisodamine decreased cardiomyocyte apoptosis, anddown-regulated mRNA levels, protein expression of the ER related caspase-12, JNKand calpain cell apoptotic pathways.4. The Ani+CA/R group showed betterarrangement of cardiomyocytes, sarcomeres and intercalated discs with higherfluorescence intensity for Cx43than the CA/R group(p<0.05).. Compared withCA/R group, the mRNA level and protein expression of Calpain in Ani+CA/R groupwere less (p<0.05), and the p-Cx43level was higher(p<0.01), is nearly to the levelof control group. However, the three groups had similar p-Cx/43/Cx43ratios.Conclusions1. Anisodamine can’t increase the ROSC ratio for electrically-induced9minutes CA. However, with anisodamine administration during CPR, the CPRtime was shorter100s than CA/R group, and ROSC ratio was increased by22.22%.2. Anisodamine can ameliorate the metabolic disorder of oxygen radicals after CAand CPR, and protects against cardiac IRI through modulation ofmitochrome-induced and endoplasmic reticulum-induced myocardium cell apoptoticsignaling pathways.3. Anisodamine protects myocardium conduction structure andfunction through up-regulation the ratio of p-Cx43/Cx43.