Intervention Of Anisodamine On Resuscitation Of Cardiac Arrest Rats And Endothelial Cells' Injury

Objective To evaluate the influence of anisodamine on the return of spontaneous circulation (ROSC) rate and resuscitation rate of cardiac arrest rats and its intervention on endothelia cells'injury so as to explore its possible protective effect and mechanism, which could be an available support for the use of Ani in cardiopulmonary resuscitation (CPR).Methods PartⅠand partⅡ:forty-five Sprague-Dawley (SD) rats were randomized to three groups:0.9% saline (control group), epinephrine group and epinephrine plus Anisodamine(Ani) group. Cardiac arrest was induced by transoesophageal cardiac pacing. A blinded drug administration and mechanical chest compression were used in the study. Return of spontaneous circulation (ROSC) rate and resuscitation rate were compared between these groups. The pathological changes of endothelia cells of brain neurons and myocardium were observed by electron microscope. Endothelin(ET) and TNF-αwere measured by radioimmunoassay. The expression of Bcl-2 was observed through immunohistochemistry methods. PartⅢand partⅣ: the study was based on human umbilical vein endothelial cell (HUVEC) line 8000.The HUVECs were divided into 5 groups:control group, anoxia/reoxygenation(A/R) 0.9% saline group, A/R epinephrine group, A/R Ani group, A/R epinephrine plus Ani group. The apoptosis of cells in each group were detected by fluorescence flow cytometry and terminal deoxynucleotide mediated nick end labeling (TUNEL). ET and NO were measured by radioimmunoassay and nitrate reduction test respectively. The gene expression of Bcl-2 and M2 receptor were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results PartⅠand partⅡ:ROSC rate, resuscitation rate and survival rate within 3 hours of epinephrine plus Ani group were higher than those of epinephrine group. Mean blood pressure (MBP) at the beginning of ROSC in epinephrine group was higher compared with epinephrine plus Ani group. Five minutes after ROSC, MBP in epinephrine group decreased qucikly than that in epinephrine plus Ani group, till thirty minutes after ROSC, MBP of epinephrine group was obviously lower than that of epinephrine plus Ani groups. The damage in neurons and myocardium of epinephrine plus Ani group were lighter than epinephrine group under electronmicroscope. In epinephrine plus Ani group, the expression of Bcl-2 was higher than that in epinephrine group. After 5min and 60min of ROSC, the level of ET of epinephrine group was higher than epinephrine group. After 60min of ROSC, the level of TNF-αof epinephrine plus Ani group were lower than epinephrine group. PartⅢand part IV:the apoptosis index(AI) of epinephrine plus Ani group were lower than epinephrine group, but higher than control group. There was no significant expression difference between epinephrine group and Ani group. In epinephrine plus Ani group, the mRNA expression of Bcl-2 was higher than epinephrine group and Ani group. In epinephrine plus Ani group, the level of ET after 12h of A/R was lower than epinephrine group, but no difference after 24h of A/R. In epinephrine plus Ani group, the level of NO after 12h of A/R was higher than epinephrine group, but no difference after 24h of A/R. The mRNA expression of M2 receptor was dectected on the HUVECs. In epinephrine plus Ani group, the mRNA expression of M2 receptor was higher than other groups.Conclusion Administration of epinephrine plus Ani at the beginning of cardiac arrest could improve ROSC rate and resuscitation rate, and could have protective effect on the injury of endothelia cells. Ani could decrease the apoptosis after A/R. Ani could protect the function of endothelia cells after A/R by effecting M2 receptor. Ani could be an important clinical drug used in CPR combined with ephrinephrine.