含二氮嗪与1，6-二磷酸果糖心麻痹液对离体大鼠心脏低温保存的实验研究
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摘要
心脏移植作为终末期心脏疾病的治疗方法已被广泛接受,而高质量的心脏保存是提高手术成功率及远期生存率的重要环节。目前临床上普遍采用的单纯低温浸泡方法安全保存心脏的时限一般在4-6 h,因此心脏保存液及保存方法仍需不断改进。心脏保存必须解决两个问题,一是减少保存期间的能量消耗,增加能量供应,维持心肌能量储备;二是减轻移植后缺血再灌注损伤。
研究发现,线粒体ATP敏感的K+通道(mitKATP)既是心肌保护的始动因素又是其效应器,MitKATP开放剂二氮嗪(DZ)对心肌缺血再灌注具有保护作用。1,6-二磷酸果糖(FDP)是细胞在糖酵解途径中产生的一个重要中间产物,可为缺氧的组织细胞提供能量来源,减轻缺氧对心肌细胞、血管内皮细胞的损害,有助于低温保存后心脏功能的恢复。目前移植器官的保存方法主要有单纯低温浸泡和持续微量灌注二种,实验证明,持续微量灌注较单纯低温浸泡能更有效地延长离体器官的保存时间。由此,我们推论附加DZ与FDP心麻痹液可能具有较好的心脏保存效果,若同时结合微量灌注方法,有可能更有效的延长心脏的保存时限。
目的:观察含DZ与FDP心麻痹液对冷保存大鼠心脏能量代谢、心肌细胞凋亡、冠脉内皮细胞结构和功能的影响,探讨其心肌保护作用的机制。同时观察微量灌注在心脏保存中的作用,探讨心脏保存方法的改进。
方法:本研究采用Langendorff离体心脏灌注模型和大鼠离体左心做功模型,分三部分进行。第一部分,SD大鼠48只,随机分为6组:STH组(St.Thomas II号液);D组(St.Thomas II号液+DZ100μmol/L);F组(St.Thomas II号液+FDP 10mmol/L);DF1组(St.Thomas II号液+DZ 25μmol/L + FDP 2.5 mmol/L);DF2组(St.Thomas II号液+DZ 50μmol/L + FDP 5 mmol/ L) ; DF3组(St.Thomas II号液+ DZ100μmol/L+ FDP 10mmol/L)。在4℃条件下保存离体心脏6h,记录保存前及保存后复灌30分钟的心率(HR)、左心室收缩压(LVSP)、左心室舒张末压(LVEDP)、左心室压力变化率最大值(±dp/dtmax)、冠脉流出量(CF)等指标的变化;并测定保存前及复灌30min时冠脉流出液中乳酸脱氢酶(LDH)及磷酸肌酸激酶(CK)的含量。第二部分:实验一,SD大鼠42只,随
Background: Heart transplantation is the most effective treatment for end-stage heart disease and has become a standard therapy in recent years. Successful heart transplantation depends on the high-quality preservation of donor hearts. Although many advances have been made in heart preservation, current hypothermic heart preservation still has many defects, one of the practical problems encountered in heart transplantation is the limited preservation time of the donor heart, which is usually 4-6 hours. How to maintain energy supply and to alleviate ischemic reperfusion injury is of great importance for heart preservation.
Recent studies suggest that: opening mitochondrial ATP-sensitive potassium channel (mitoKATP) has cardioprotective effects on ischemia-reperfusion injury. As an important intermediate product of glycolytic pathway, Fructose-1,6-diphosphate(FDP) can increase energy supply of ischemic cells, alleviate ischemia injury, and improve function recovery of the heart. Microperfusion has been shown to be a promising technique for heart preservation. Therefore, we suppose that adding DZ and FDP to St.Thomas II solution, would improve the effects of heart preservation, and prolong the storage time limit combining with microperfusion technique.
Objective: To investigate the protective effects of St. Thomas II solution supplemented with DZ and FDP on myocardial energy metabolism, myocardial apoptosis and endothelial cell injury of hypothermic preservative rat heart; To evaluate the effects of the new solution combining with microperfusion technique on prolonging the storage time.
Methods: Isolated rat heart Langendorff model and working model were established. The study consisted of three parts: In the first part, 48 SD rats were divided into six groups randomly: STH group (St.Thomas II solution); D group (St.Thomas II solution +
研究发现,线粒体ATP敏感的K+通道(mitKATP)既是心肌保护的始动因素又是其效应器,MitKATP开放剂二氮嗪(DZ)对心肌缺血再灌注具有保护作用。1,6-二磷酸果糖(FDP)是细胞在糖酵解途径中产生的一个重要中间产物,可为缺氧的组织细胞提供能量来源,减轻缺氧对心肌细胞、血管内皮细胞的损害,有助于低温保存后心脏功能的恢复。目前移植器官的保存方法主要有单纯低温浸泡和持续微量灌注二种,实验证明,持续微量灌注较单纯低温浸泡能更有效地延长离体器官的保存时间。由此,我们推论附加DZ与FDP心麻痹液可能具有较好的心脏保存效果,若同时结合微量灌注方法,有可能更有效的延长心脏的保存时限。
目的:观察含DZ与FDP心麻痹液对冷保存大鼠心脏能量代谢、心肌细胞凋亡、冠脉内皮细胞结构和功能的影响,探讨其心肌保护作用的机制。同时观察微量灌注在心脏保存中的作用,探讨心脏保存方法的改进。
方法:本研究采用Langendorff离体心脏灌注模型和大鼠离体左心做功模型,分三部分进行。第一部分,SD大鼠48只,随机分为6组:STH组(St.Thomas II号液);D组(St.Thomas II号液+DZ100μmol/L);F组(St.Thomas II号液+FDP 10mmol/L);DF1组(St.Thomas II号液+DZ 25μmol/L + FDP 2.5 mmol/L);DF2组(St.Thomas II号液+DZ 50μmol/L + FDP 5 mmol/ L) ; DF3组(St.Thomas II号液+ DZ100μmol/L+ FDP 10mmol/L)。在4℃条件下保存离体心脏6h,记录保存前及保存后复灌30分钟的心率(HR)、左心室收缩压(LVSP)、左心室舒张末压(LVEDP)、左心室压力变化率最大值(±dp/dtmax)、冠脉流出量(CF)等指标的变化;并测定保存前及复灌30min时冠脉流出液中乳酸脱氢酶(LDH)及磷酸肌酸激酶(CK)的含量。第二部分:实验一,SD大鼠42只,随
Background: Heart transplantation is the most effective treatment for end-stage heart disease and has become a standard therapy in recent years. Successful heart transplantation depends on the high-quality preservation of donor hearts. Although many advances have been made in heart preservation, current hypothermic heart preservation still has many defects, one of the practical problems encountered in heart transplantation is the limited preservation time of the donor heart, which is usually 4-6 hours. How to maintain energy supply and to alleviate ischemic reperfusion injury is of great importance for heart preservation.
Recent studies suggest that: opening mitochondrial ATP-sensitive potassium channel (mitoKATP) has cardioprotective effects on ischemia-reperfusion injury. As an important intermediate product of glycolytic pathway, Fructose-1,6-diphosphate(FDP) can increase energy supply of ischemic cells, alleviate ischemia injury, and improve function recovery of the heart. Microperfusion has been shown to be a promising technique for heart preservation. Therefore, we suppose that adding DZ and FDP to St.Thomas II solution, would improve the effects of heart preservation, and prolong the storage time limit combining with microperfusion technique.
Objective: To investigate the protective effects of St. Thomas II solution supplemented with DZ and FDP on myocardial energy metabolism, myocardial apoptosis and endothelial cell injury of hypothermic preservative rat heart; To evaluate the effects of the new solution combining with microperfusion technique on prolonging the storage time.
Methods: Isolated rat heart Langendorff model and working model were established. The study consisted of three parts: In the first part, 48 SD rats were divided into six groups randomly: STH group (St.Thomas II solution); D group (St.Thomas II solution +
引文
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