The Effect Of Anisodamine On Cardiac Arrest Rat And Hypoxia/Reoxygenation Cardiomyocyte

According to some researches in CPR,the increase of dose of epinephrine could increase cardiac and cerebral perfusion pressure and blood perfusion flow,but it could also do damage to the myocardia and the brain tissues,resulting in the decrease success rate of CPR.anisodamine can increase the coronary blood flow,improve the microcirculation,reduce the aggregation of platelet,prevent the thrombosis,and alleviate the ischemia-reperfusion injury of the myocardia and the brain tissues,etc.In the early stage of CPR the application of anisodamine may improve tissue perfusion, improve the function of cardiac and cerebral tissues,reduce the side-effect of epinephrine,increase the CPR success rate and improve the prognosis.Therefore,the function of anisodamine in CPR was experimented and studied.Objective To establish a standard rats' cardiac arrest and CPR model for preclinical medicine research,using blind method to deliver the medicine,observe anisodamine's effect on functions of myocardia and brain tissues of rats which suffers from cardiac arrest during CPR,and further observe anisodamine's protection effect on hypoxia/reoxygenation cardiomyocyte in terms of apoptosis.Methods There were four parts in the experiment.In partⅠ:Thirty Sprague-Dawley(SD) rats were randomly devided 2 groups,the physiological saline control group(n=15) and epinephrine group(n=15).The cardiac arrest model of rats were induced by alternating current(AC) via pacing electrode placed in esophagus. Five minutes after onset of cardiac arrest,CPR was initiated.Cardiac compression was performed by machinery which was made by our group.The types of cardiac arrest:vntricular fibrillation(VF),pulseless electrical activity(PEA),asystole were inspected,duration of the pace,restored of self circulation(ROSC) and success resuscitation were also detected.Necropsy of the esophaguses,the hearts and the organs of the abdominal were performed after the test to see whether the esophagus or the heart were burned and whether the internal organs were broken.In partⅡ65 SD rats were randomly divided to physical group(n=5),physiological saline group(n=15), epinephrine group(n=15),anisodamine and epinephrine combination group(n=15), anisodamine(Ani) group(n=15).We injected 0.6ml FDG before the stimulation.The cardiac arrest rates modle were created and treated as partⅠ.Medicine was given in blind way.The hearts and brain of the rats were removed for radiocounting,and the plasma A-type natriuretic peptide(ANP) were checked.In partⅢ:On the base of partⅡ,apoptosis of cardiamyocytes and brain cells were identified as DNA fragmentation by TUNEL.Mitochondrion were detected by electron microscope.In partⅣwas cell culture study.The ventricular muscle cells of suckling mice were cultured,and after one generation they were divided into 5 groups,normal physiological control group, hypoxia/reoxygenation group,epinephrine group,epinephrine and anisodamine group, anisodamine group,each group hypoxia 2 hours/reoxygenation 12 hours.In each group there were at least 3 bottles of cells.By means of flow cytometry the cells' apoptosis was checked after hypoxia/reoxygenation.Meanwhile,TUNEL method was used to check the non-viable apoptotic cells and the RT-PCR method was used to check the myocardia cell M1-3 receptor subtype expression.Results 1.All of the rats were induced into cardiac arrest,VF(n=10,33.3%), PEA(n=15,50%),asystole(n=5,16.7%).Duration of the pacing was 60s to 150s. RSOC and success resuscitation rates of the controll group and the epinephrine group were(6.7%vs 0) and(46.6%vs 33.3%) respectively.Necropsy showed that there was no gross evidence of thermal injury on the surface layer of the heart,only mild to moderate thermal injury of the esophagus and no perforation,no rupture of the belly organ.2.The recovery rate of Ani and Epi combination group was higher than that of Epi group and Ani group.The myocardium and the cerebrum radiocounting of the Api recovery group were lower than those of the Ani and Epi combination recovery group, which of the myocardium were(25.71±2.83 vs 38.3±7.2,P＜0.05) and the cerebrum were(16.06±2.93 vs 31.02±2.72,P＜0.05),respectively.The plasma ANP level of the recovery in the Ani and Epi combination group and the Ani group were lower than that of the Epi group(1963.1±1096.7,483.6±205.9 vs 4417.6±2235.9,P＜0.05),but all of these groups were higher than that of the physical group(238.3±190.0,P＜0.05).3.The apoptotic cell percentage of brain tissues of surviving rats in Epi group was much higher than that in normal physiological group and Epi and Ani combination group, which was(8.85±1.2 vs 0.87±0.25,4.25±1.05,P＜0.01).The apoptotic cell percentage of myocardia of surviving rats in Api group was much more than that in normal physiological group and Epi and Ani combination group,which was(8.15±1.60 vs 2.13±0.65,4.07±1.55,P＜0.01).The injury of brain and myocardia mitochondria of rats which experience successful CPR in Epi and Ani combination group was much milder than that in Epi group.4.The cell culture:Reoxygenation 12 hours later,the early apoptotic cells of Epi group were much higher than those of the other groups (P＜0.01).The early apoptotic cells of Api group and Epi and Ani combination group were lower than those of hypoxia/reoxygenation group.The Non-viable apoptotic cells of epinephrine group was much higher than those of Ani group(P＜0.01),and lower than those of hypoxia/reoxygenation group and Epi and Ani combination group, but there's no sense in terms of statistics.RT-PCR checks the myocardia cell M2 receptor subtype expression,which had no statistic differences among the groups.Conclusions 1.A good cardiac arrest model of rats can induced by alternating current via pacing electrode placed in esophagus.Cardiac compression performed by machinery can give a standard CPR research in the rat.2.Anisodamine is useful in CPR,and can improve metablism ofmyocadium and cerebrum.3.Anisodamine may decrease the apoptosis in hypoxia/reoxygenation myocardium cell,so as to protect the hypoxia/reoxygenation myocardium cell.