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Activation of ERK during ischemia-reperfusion in lung transplantation.

肺移植缺血再灌注过程中 ERK 的激活。

基本信息

批准号：
14571268
负责人：
SAKIYAMA Shoji
金额：
$ 2.18万
依托单位：
THE University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

1.Experiment in a rat lung transplant model.Dramatic alterations of protein tyrosine phosphorylation have been found during the ischemia-reperfusion period of human lung transplantation. The object of the present study was to determine whether these changes exist in a rat single-lung transplant model. Lung transplantations were performed after 6 hours of cold ischemic preservation. In both iso-and allografts, the lung function of transplants was very well preserved. Protein tyrosine phosphorylation, protein tyrosine kinase and protein tyrosine phosphatase activities were decreased significantly after 2 hours of reperfusion. Src protein level and phosphorylation of p38 were reduced after 2 hours of reperfusion. This lung transplantation model is useful for studying ischemia-reperfusion injury.MAPK activation in human lung transplantThe phosphorylation status of MAPK during ischemia-reperfusion in human lung transplant was examined. These transplantations were performed in Toronto General Hospital in Canada. Lung tissue biopsy was performed on 15 patients undergoing transplantation: after the cold ischemic preservation, after the warm ischemia (implantation) and after 1 h or 2 h reperfusion. The phosphorylation status of ERK, p38-MAPK, INK and MEK was examined with Western blotting. Phosphorylation of INK also significantly increased at lower levels. In contrast, phosphorylation of p38 had no significant changes. The increase in ERK phosphotylation was not associated with the activation of MEK, the up-stream kinase for ERK. Since MKP-3 protein levels correspond to its activity, we examined the protein expression of MKP-3 in these samples. Surprisingly, the MKP-3 protein level significantly increased after reperfusion. Therefore, the dramatic increase in ERK phosphorylation is not due to the inhibited expression of MKP-3.

1.大鼠肺移植模型实验。在人肺移植缺血再灌注期，蛋白酪氨酸磷酸化发生了显著变化。本研究的目的是确定这些变化是否存在于大鼠单肺移植模型中。冷缺血保存6小时后进行肺移植。在同种异体和同种异体移植中，移植的肺功能都得到了很好的保存。蛋白酪氨酸磷酸化、蛋白酪氨酸激酶和蛋白酪氨酸磷酸酶活性在再灌注2 h后显著降低。再灌注2小时后，Src蛋白水平和p38磷酸化水平降低。该肺移植模型可用于研究缺血再灌注损伤。研究MAPK在人肺移植缺血-再灌注过程中的磷酸化状态。这些移植是在加拿大多伦多总医院进行的。对15例移植患者分别在冷缺血保存后、热缺血（植入）后、再灌注1 h或2 h后进行肺组织活检。Western blotting检测ERK、p38-MAPK、INK和MEK的磷酸化状态。在较低水平下，INK的磷酸化也显著增加。相比之下，p38的磷酸化没有明显变化。ERK磷酸化的增加与ERK的上游激酶MEK的激活无关。由于MKP-3蛋白水平与其活性相对应，我们检测了这些样品中MKP-3蛋白的表达。令人惊讶的是，再灌注后MKP-3蛋白水平显著升高。因此，ERK磷酸化的急剧增加并不是因为MKP-3的表达受到抑制。