【作者】 孟辉；

【导师】 陈钢；

【作者基本信息】 第一军医大学 ， 外科学（普胸专业）， 2001， 硕士

【摘要】 研究目的 在分析ECS及MECS的基础上修改配方成分自制IECS；探讨犬肺低温灌洗、离体保存过程中随着冷缺血时间推移NO、MDA、SOD的变化规律；探讨分子水平肺组织活性改变与形态学变化的相互关系以及体视学量化原理在供体肺研究中的可适用性。材料和方法 12只雄性健康杂种犬随机分入供体组和受体组，供体组之供肺以灌洗前为对照组；灌洗后低温保存期间不同时间点为实验观察组。麻醉生效后，供体组：开胸后经肺动脉插管注入PGE₁400μg，采用低温单次原位顺行法灌洗4℃ IECS 1500ml，压力控制在3～4kPa。灌洗完毕后完整摘取心肺，一侧肺作为供肺移植，另一侧肺离体低温（4℃）保存，测试、观察不同时间点肺组织分子生化和形态学改变，包括：肺组织W/D；制备匀浆，测定肺组织内NO、MDA和SOD浓度；肺组织形态学变化的定性观察及描述；用方网测试系统体视学量化原理测定肺泡上皮细胞核/质Vv、Sv、Rsv。受体组：开胸后行单侧全肺切除并修整残端，待移植肺残端修整后与之吻合；移植前、中、后监测血流动力学及呼吸各项参数指标。结 果 实验组与对照组相比较：W/D，NO、SOD和MDA含量，肺泡内出血分级，肺泡上皮细胞核/质Vv、Sv、Rsv随低温保存时间变化无显著性差异（P＞0.05）；但随时间延长NO、SOD、Vv、Sv均呈下降趋势，而MDA、W／D、RSV却略有上升趋势。保存期间细小支气管上皮细胞和血管内皮细胞结构正常，肺组织、肺泡内出血变化不明显，主要改变是组织水肿。结 论 1艰据犬肺离体低温保存后肺组织Wto，NO、SOD和 MDA含量，肺泡内出血分级，肺泡上皮细胞核／质VV、SV、RSV随时间延长变化的特点，可见肺组织病理形态学改变略滞后于OFR变化，随着保存时间延长肺组织水肿渐趋明显。 2在本实验条件下所采用的IECS进行犬肺灌洗、离体保存4h之内肺血管内皮细胞无明显损伤，PGE；具有保护肺血管内皮细胞的作用；甘露醇具有增加渗透压和清除自由基的作用；灌洗前肺动脉内注射PGE；及增加 ECS渗透压和代谢底物、提高PH值使之略微偏碱均有助于抑制供体肺离体、低温、冷缺血保存期间OFR的产生。 3．VV、SV、RSV作为肺泡上皮细胞核／质形态学变化的量化指标，体视学定量原理在供肺保存质量研究中具有实用性。更多还原

【Abstract】 OBJECTIVES The purposes of this study were (I) to investigate improved Euro-Collins solution (IECS) after analyzing the ingredient of Euro-Collins solution (ECS) and modified Euro-Collins solution (MECS), (2) to study on the variation of NO, MDA, SOD and the variety of morphology in isolated canine donor lung followed by cold ischemic time, (3) to evaluate the applicability of quantized principle of Biological Stereology in research of donor lung and to provide experimental basis for observing in the model of the donor lung. MATERALS AND METHODS Twelve adult mongrel canines of male, weighing 19-24kg, anesthetized with sodium pentobarbital, were randomly assigned to donor group (n=6) and recipient group (n=6). The donor group was divided into control and experiment group still further before and after perfusion. After the pulmonary artery of donor injected by prostaglandin E1 (POEI, 4OO~.tg), the lung was flushed by IECS (1 500ml) with direct single flush perfusion (SFP) at 4 0C and pressure in the master of 3-4 kPa. Then to take out the whole of heart and lungs, one of the lungs was for transplantation and the other stored for determining the changes of molecule biochemistry and morphology involving W/D and the concentrations of NO, SOD, MDA at various time. The quantity stereological parameters of the nucleolus /cytoplasm volume density (Vv) , surface density (Sv) and ratio of surface to volume for X phase (Rsv) of the alveolar epithelia cells were measured and characterized by the 4 square net testing system of Biological Stereology. Recipient group: one side of total pneumonectomy was performed, and then, the single lung transplantation was finished at the same side. The circulation and respiration functional parameters were monitored before, during and after transplantation. RESULTS Comparison of control and experimental groups, no differences were found in W/D, NO, SOD, MDA, bleeding grade in alveolus and Vv, Sv, Rsv of the nucleolus /cytoplasm in the alveolar epithelia cells (P> 0.05), but there were the downtrends in NO, SOD and Vv, Sv, and the uptrends in MDA, W/D and Rsv along with prolonging time. The main changes were not haemorrhage in tissues and alveolus of donor lung, but the pulmonary edema was existent in there. CONCLUSIONS 1. The characteristics of W/D, NO, SOD, MDA, bleeding grade in alveolus, Vv, Sv and Rsv in the nucleolus /cytoplasm of the alveolar epithelia cells in the isolated donor lung showed that the morphological changes were later than OFR. The pulmonary edema was gradual evidence along with prolonging time. 2. In this experimental conditions, the VEC had not damnified obviously by perfusion with IECS in 4 hours. The pulmonary vascular resistance could decreased by POE1 and so vascular endolhelial cells would be protected. Mannitol not only increase osmotic pressure but also cleanse OFR. The addition of mannitol, POE1, glucose and osmotic pressure to IEC solution contributes to reduce the quantities of OFR. 3. The quantity stereological parameters of the nucleolus /cytoplasm Vv and Sv and Rsv of the alveolar epithelia cells were the measure parameters of alveolar epithelia cells. Biological Stereology may be applied to the experimental study of lung preservation.更多还原