乌司他丁减轻无心跳大鼠供肺缺血再灌注损伤的作用及机制

Protective effects of ulinastatin on ischemia-reperfusion injury during rat non-heart beating donor lung transplantation

目的研究乌司他丁（UTI）减轻无心跳大鼠供肺缺血再灌注损伤作用及作用机制。方法选取雄性SD大鼠作为供、受鼠建立无心跳大鼠左肺移植模型，将受鼠分为对照组和实验组，对照组受鼠接受的供肺经低钾右旋糖酐（LPD）液灌注和保存，实验组受鼠接受的供肺经含乌司他丁（50万U／L）的LPD液灌注和保存。移植过程中监测受鼠的动脉血氧合情况；移植肺再灌注30min和1h时，取两组受鼠移植肺组织，测量和计算湿干质量比，检测移植肺组织中丙二醛（MDA）和超氧化物歧化酶（SOD）含量；提取RNA，采用实时定量聚合酶链反应检测移植肺组织中肿瘤坏死因子-a（TNF-d）、细胞问黏附分子-1（ICAM-1）和白细胞介素10（IL-10）mRNA的相对表达量。结果再灌注后1h，实验组的氧合指数（Pa02／Fi02）为472．38±31．66，显著高于对照组的429．52±14．83，两组比较，差异有统计学意义（P=0．025）。与对照组相比，实验组在再灌注30min和1h时的水肿情况（湿干质量比）均好于对照组（P=0．005，P=0．006），实验组移植肺组织病理损伤也明显轻于对照组。不论再灌注30min还是1h，实验组移植肺组织中MDA含量较对照组显著降低（P=0．039，P=0．006），而SOD含量显著升高（P=0．035，P=0．030）。再灌注30min时，实验组TNF-a“的表达较对照组显著下降（P=0．000），再灌注1h时下降不明显（P=0．139）；再灌注30min时，ICAM-1水平较对照组下降不明显（P=0．062），再灌注1h则存在明显降低（P=0．001）；再灌注30rain和1h时，实验组IL-10mRNA的表达水平均较对照组显著上调（P=0．004，P=0．000）。结论乌司他丁能够减轻无心跳大鼠供肺的缺血再灌注损伤，对移植肺有保护作用。

Objective To investigate the protective effects of ulinastatin （UTI） on ischemi reperfusion injury of donor lungs, and the possible mechanism. Method Forty male SD rats were randomly divided into two groups： group C as control group and group U as UTI group. In group C donor lungs were antegradely flushed with 20 ml of cold （4 C） low potassium dextron （LPD） solution and 5 ml retrogradely. Meanwhile, in group U, UTI （500 000 U/L） was added in LPD solution and the same doses were used. According to the time after initiation of reperfusion, each group was divided into two subgroups： 30 rain （subgroup A） and 1 h （subgroup B）. Arterial blood samples were collected for blood gas analysis. Lung samples were obtained at the end of reperfusion （30 rain or 1 h）. Microscopic examination of the donor lungs was conducted. Besides, the pulmonary water index （W/D）, tissue malondialdehyde （MDA） and superoxide dismutase （SOD） content, and mRNA expression of tumor necrosis factor （TNF-a）, intercellular adhesion molecule 1 （ICAM 1） and interleukin 10 （IL-10） were also measured. Results （1） One h after reperfusion, oxygenation index in group U was higher than that in group C （P = 0. 025） ; （2） The levels of W/D in subgroup A and subgroup B of group U were decreased as compared with group C （P = 0. 005 and P = 0. 006） ; （3） The microscopic changes of donor lung tissues in group U were lessen than in group C; （4） In subgroup A of group U, MDA content was decreased （P = 0. 039）, and SOD content was increased （P= 0. 035）, and similar results could be observed in subgroup B of group U （P = 0. 006 and P = 0. 030 respectively） （5） As compared with group C, the mRNA expression of TNF-a in group U was decreased at the time of 30 min after reperfusion （P = 0. 000）, but no significant change was found at the time of 1 h （P = 0. 139）. The mRNA expression of ICAM-1 was not decreased evidently at the time of 30 min （P = 0. 062）, but significantly decreased at the time of 1 h （P = 0. 001）. The mRNA expression of IL-10 was increased in subgroups A and B （P = 0. 004 and P = 0. 000 respectively）. Conclusion This study demonstrated that UTI had protective effects of reducing ischemia-reperfusion injury on the donor lungs after lung transplantation in rat non-heart beating： donor models.