大鼠低温假死-延时复苏动物模型建立

Establishment of hypothermia-induced suspended animation for delayed resuscitation animal model in rats

目的探讨通过全身高钾诱导心脏停搏、停循环建立大鼠低温假死-延时复苏动物模型的可行性。方法选取SPF级健康雄性SD大鼠6只,按照大鼠体外循环动物模型进行插管准备并放血建立失血性休克模型。而后通过动脉灌注管持续灌注冷康思德脏器保护液(HTK液)诱导心脏停搏并全身降温,当温度降至15℃时停循环。停循环60 min后,恢复血液灌注并体外循环辅助至恢复稳定正常心率。结果6只大鼠在体外循环维持下均可维持稳定的自主心率,仅1只大鼠可脱离体外循环(体外循环时间152 min)。建立休克模型期间放血(4.2±0.8)ml,诱导低温假死阶段应用HTK液(135±14)ml,从HTK液灌注到体温降至15℃停循环耗时(27±4)min,恢复血液灌注后体内K+浓度(8.9±4.3)mmol/L,延时复苏阶段输注同种异体血(11±1)ml。乳酸值在休克诱导后开始逐渐升高,在恢复循环10 min后达到高峰,随着循环恢复逐渐出现下降趋势。结论本研究通过全身灌注HTK溶液成功建立大鼠低温假死-延时复苏动物模型,为研究低温假死-延时复苏对脑、心、肝、肾等重要脏器的损伤及保护机制奠定基础。

Objective To investigate the feasibility of rats model of hypothermia-induced suspended animation for delayed resuscitation which were carried out cardioplegia and circulatory arrest induced by systemic hyperkalemia.Methods Six healthy male Sprague-Dawley rats with SPF grade were selected to cannulate according to the cardiopulmonary bypass rat model,and bled to establish the hemorrhagic shock model.Then,cold organ preservation solution(HTK solution)was continuously perfused to induce cardioplegia and systemic hypothermia through the artery catheter,and circulatory arrest was induced when the core temperature dropped to 15℃.Hemoperfusion was restored after 60 minutes of circulatory arrest until restoration of stable and normal heart rate.Results Six rats could maintain stable spontaneous rate with cardiopulmonary bypass,but only one rat could wean off cardiopulmonary bypass(cardiopulmonary bypass time:152 min).The rats were bleeding(4.2±0.8)ml during the establishment of shock model.During hypothermia-induced suspended animation,the consumption of HTK solution was(135±14)ml,and the induction time from HTK solution perfusion to temperature dropping to 15℃was(27±4)min.After reperfusion,the concentration of potassium ion was(8.9±4.3)mmol/L in vivo.The volume of allogeneic blood transfusion was(11±1)ml during delayed resuscitation.The lactate levels gradually rised after induction of shock,reached to the peak at 10 minutes after restoration of circulation,and gradually showed a downward trend with the extension duration of cardiopulmonary bypass support.Conclusions Systemic perfusion with organ preservation solution(HTK solution)can establish rats model of hypothermia-induced suspended animation for delayed resuscitation,and this method laids the foundation of the study on damage and protection mechanism of hypothermia-induced suspended animation for delayed resuscitation on brain,heart,liver,kidney and other important organs.