【作者】 段大为；

【导师】 刘维永；

【作者基本信息】 第四军医大学 ， 外科学， 2004， 硕士

【摘要】 目的： 研究药物预处理及在改良的St.Thomas Ⅱ液中加入钾离子通道开放剂和阿片受体激动剂对离体心脏的保存效果。以及尼可地尔再灌注对减轻钙离子超载的作用。为改善供心保存寻求更有效的方法。 方法： 1．利用改良的Langendorff灌注装置，建立离体心脏灌注模型。 2．在ST，Thomans Ⅱ液中加入尼可地尔作为实验组Ⅰ组的离体心脏保存液(钾离子浓度8.0mmol／L)。观察药物预处理及低温保存不同时间和再灌注30min后离体心脏的恢复情况；对照组用UW液作为离体心脏的保存液； 3．在尼可地尔预处理和低温下保存8h的基础上，在K-H灌注液中加入尼可地尔，观察其保护效果。与尼可地尔预处理和低温保存8h组相比较。 4．在ST，Thomans Ⅱ液中加入U50，448H作为实验组Ⅱ组的离体心脏保存液(钾离子浓度8.0mmol／L)。观察药物预处理及低温保存不同时间和再灌注30min后离体心脏的恢复情况；与尼可地尔预处理低温保存组比较。观察二者对离体心脏保存作用的差异。 5．实验动物大白兔在麻醉前分别用尼可地尔或阿片受体激动剂U50，488H预处理。 6．实验动物依据离体供心的保存时间不同分为4个时间点进行观察：保存4h，6h，8h，10h。尼可地尔预处理和低温保存+尼可地尔再灌注组只观察保存8h的心脏变化。 7．记录冷灌前心功能指标。测定再灌注30min时离体心脏的左室发展压(DP)和左室压力微分(±dp／dt)、冠状动脉流量(CF)、并计算心功能恢复百分比，留取冠状动脉引流液测定心肌酶(LDH、CK)、心肌线粒体钙含量、肌浆 举四军反大掌硕士研究生毕业论文亘里旦口里国.里口口口里里里国旦里旦里里鱼旦旦里皿旦旦鱼旦旦旦旦鱼旦旦旦旦旦鱼鱼鱼旦里里旦鱼鱼鱼旦里旦~ 网ca2+一ATPase活性、心肌细胞ATP含量，心肌组织含水量，并观察心肌 超微结构。结果:1.尼可地尔预处理低温保存组:离体心脏在经尼可地尔预处理低温保存后随 着保存时间的延长钙离子仍有明显的超载，实验组与对照组相比心功能恢 复率及钙离子超载程度在保存6h内的结果差异性不显著(p>0.05);在 sh后二个时间点的钙离子超载程度结果差异性明显(p<0.01)。2.离体心脏在经尼可地尔预处理和低温保存后对Caz+一ATPase活性有保护作 用;实验组和对照组CaZ十一ATPase活性在相应的时间点存在明显差异(P< 0.01);两组线粒体钙离子含量在8h组存在明显差异(P<0.05)。3.尼可地尔预处理可以明显改善心肌结构的改变;Uw液对冠脉血管内皮损 伤严重，使心脏再灌注后冠状动脉流量恢复率明显减少4.尼可地尔预处理和低温保存+尼可地尔再灌注可以进一步减轻钙离子的 超载，减轻心肌结构的变化，但心功能的恢复较尼可地尔预处理和低温保 存组降低，且有明显差异(p<0.05)。5.U50，4 88H预处理和低温保存离体心脏可以减轻钙离子超载;实验组不同 保存时间点的组间比较，钙离子超载程度、Ca2+一ATPase活性、ATP含量 等相比较存在明显的差异。但与对照组相比较，钙离子的超载程度、 ca2+一ATPase活性在不同的保存时间点，没有明显的差异(p>0.05)。6.阿片受体激动剂U50488H和钾离子通道开放剂尼可地尔对离体心脏预处 理和低温保存作用没有明显的差异性结论:1.尼可地尔预处理和低温保存离体心脏可以减少心肌细胞的钙离子超载， 减轻缺血再灌注损伤.供心保存时间在6h内，与Uw液的保存效果相当。 在sh组UW液的保存效果较尼可地尔预处理和低温保存差2.改良ST，ThomanSH(K+浓度8n’uno1/L)液可以使心脏平稳的停跳，砂自脏停 跳所需时间较高钾的Uw液有所延长，但差异性不显著3.高钾的Uw液对心脏血管内皮有明显的损伤。并且使心肌组织的水肿程度 增加。注.尼可地尔再灌注可以进一步减轻钙离子超载，减轻缺血再灌注损伤，但举四军反大学硕士研究生华业论文 对心功能的恢复有一定的负面影响。尼可地尔再灌注与尼可地尔预处理 对心脏结构保存和保护有协同作用。5.阿片受体激动剂U50，488H预处理和低温保存对离体心脏有一定的保护作 用，其作用和钾离子通道开放剂尼可地尔作用效果基本相同。更多还原

【Abstract】 ObjectiveTo study the action of Pretreatment to isolated rabbit hearts with the Potassium ionic channel opener Nicorandil and the opium recepter Excitation opener U50588H by improve Langendorff perfusion model. The preservation action was observed for the Nicorandil to re-perfuse the isolated rabbit hearts to relieve calcium ions overloading and to preserve time. To observed the preservation effect for isolated rabbit hearts with improved St.Thomas II solution, to explore a new operative preservation way for the isolated heart in clinical heart transplantation. Method1. The Langendorff perfusion assembly was used to set up perfusion model for isolated hearts.2. The Nicorandil was added into ST,Thomans II solution as isolated hearts’ preservation solution for study group I (K+ was 8mmol/L). To observe the variation of Cardiac function restoration, cardiac muscle cell calcium ions, myocardium Adenosine triphosphate (ATP), myocardium enzyme, the water in myocardium, myocardium ultrastructure, et al. The University of Wisconsin solution (UW) as a kind of preservation solution was used for isolated hearts in control group.3. On the base of Nicorandil Pretreatment and Nicorandil solution to hypothermia preserve the isolated hearts for 8h, moreover added the Nicorandil into the Krebs-Hensleit solution to perfuse the isolated rabbit hearts, to observe its protection effect for isolated rabbit hearts. The control group that was Pretreatment by Nicorandil and Nicorandil solution to preserve in hypothermia the isolated hearts for 8h.4. The U50488H was added into ST.Thomans II solution as a kind of isolatedhearts’ preservation solution for study group II (K+ was 8mmol/L). To observe the variation of Cardiac function restoration, cardiac muscle cell calcium ions, myocardium ATP, myocardium enzyme, and the water in myocardium, myocardium ultrastructure, et al. To compare with the different for the isolated rabbit hearts between in this group and Nicorandil Pretreatment group.5. Experiment animals the big white rabbits were pretreatment by Nicorandil or U50,488H before anesthetized.6. According to the different preservation times for the isolated rabbit hearts, the experiment animals were divided into four time points to observe: four hours; six hours; eight hours and ten hours. About the Nicorandil re-perfusion, we only observed the isolated hearts that were preserved for eight hours group.7. Read-in the Cardiac function markers about the isolated hearts before them were perfuse the cardioplegia solution. To assay the left ventricular developed pressure (LVDP), left ventricular pressure differential and integral calculus (眃p/dt), coronary artery flow (CF) and to compute the percentage for Cardiac function recover; Assay the myocardium enzyme (lactate dehydrogenase, LDH; the cardiac muscle phosphokinase, CK) from the coronary artery drainage solution , the Content of calcium in myocardium mitochondrion, sarcoplasmic reticulum Ca2+-ATPase activity, ATP content in myocardium, myocardium water content and to obverse the myocardium ultrastructure.Result 1. There wasn’t obviously difference for the rate of the cardiac function recover between test group and control group when the preservation time less then six hours (P>0.05) ,but the rate of the cardiac function recover there was obviously difference in two groups(p<0.05); at ten hours the cardiac function recover there wasn’t obviously difference (P > 0.05) .2. Nicorandil Pretreatment could safeguard the Ca2+-ATPase activity, meanwhile it could relieve calcium ions overloading. About the Ca2+-ATPase activity at the same time there was obviously difference between test group and controlgroup(p<0.01); After eight hours, the mitochondrion calcium ions there were obviously difference (p<0.05).3. Nicorandil Pretreatment could obviously improve myocardium structural; UW solution could cause Severity injury for coronary artery endothelium, this made the recovery rate of the corona更多还原