Activations of Mitogen-Activated Protein Kinases and Regulation of Their Downstream Molecules After Rat Lung Transplantation from Donors After Cardiac Death
Activations of Mitogen-Activated Protein Kinases and Regulation of Their Downstream Molecules After Rat Lung Transplantation from Donors After Cardiac Death
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DOI：
10.1016/j.transproceed.2011.09.075
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发表时间：
2011-12-01
影响因子：
0.9
通讯作者：
Miyoshi, S.
中科院分区：
文献类型：
--
作者：
Yamamoto, S.;Yamane, M.;Miyoshi, S.
目标.接受心脏性死亡后器官捐献是解决供体短缺的有效途径。然而，来自DCD供体的肺移植显示严重的快速肺移植物功能障碍（PGD），随后是热缺血再灌注损伤（IRI）。本研究旨在阐明温热IRI的分子介质，包括丝裂原活化蛋白激酶（MAPK）的激活及其下游通路。在热缺血时间为0分钟（CIT组）、30分钟（30 WIT组）或180分钟（180 WIT组）后，我们使用雄性Sprague-Dawley大鼠的器官进行单左肺移植。通过血气分析评估移植肺功能。再灌注后1小时，采用蛋白质印迹和实时聚合酶链反应检测移植物中MAPKs（ERK、p38和JNK）的磷酸化状态以及转录因子（Egr-1和ATF-3）和免疫介质（MCP-1、MIP-2、派-1、ICAM-1、TNF-α、IL-1 β、IL-6和考克斯-2）的基因表达水平。结果。与其他组的移植肺相比，在180 WIT组中观察到严重的PGD，其表现出良好的肺移植功能。ERK和JNK的激活，以及转录因子（Egr-1和ATF 3）的mRNA水平显着增加与更大的热缺血时间。JNK激活模式与PGD的严重程度相关。MCP-1、ICAM-1、IL-1 β、IL-6和考克斯-2在180 WIT组中也表达上调，但MIP-2和派-1在各组间无显著差异。我们认为ERK和JNK通路可能在DCD供体肺移植中诱导长期热缺血再灌注损伤中起重要作用。
Objectives. Accepting organs donated after cardiac death (DCD) is an effective approach to the donor shortage. However, lung transplantations from DCD donors show severe rapid pulmonary graft dysfunction (PGD) followed by warm ischemia-reperfusion injury (IRI). This study sought to clarify the molecular mediators in warm IRI, including activation of mitogen-activated protein kinase (MAPK) and the downstream cascades.Methods. We performed single left lung transplantation using organs from male Sprague-Dawley rats after 0 (CIT group), 30 (30WIT group), or 180 (180WIT group) minutes of warm ischemia time. Pulmonary graft functions were estimated by blood gas analysis. At 1 hour after reperfusion, the phosphorylation status of MAPKs (ERK, p38, and JNK) and the gene expression levels of transcription factors (Egr-1 and ATF-3) and immune mediators (MCP-1, MIP-2, PAI-1, ICAM-1, TNF-alpha, IL-1 beta, IL-6, and COX-2) in the grafts were examined using Western blotting and real-time polymerase chain reaction assays.Results. Severe PGD was observed in the 180WIT group compared with transplanted lungs in the other groups, which exhibited good pulmonary graft function. ERK and JNK activations, as well as mRNA levels of transcription factors (Egr-1 and ATF3) significantly increased with greater warm ischemic times. The pattern of JNK activation correlated with the severity of PGD. MCP-1, ICAM-1, IL-1 beta, IL-6, and COX-2 were also up-regulated among the 180WIT group, although MIP-2 and PAI-1 showed no significant differences among the groups.Conclusions. We suggest that the ERK and JNK pathways may play important roles to induce the injury caused by prolonged warm ischemia followed by reperfusion in the setting of lung transplantation from DCD donors.