Gene transfer of heat shock protein 70 protects lung grafts from ischemia-reperfusion injury
Gene transfer of heat shock protein 70 protects lung grafts from ischemia-reperfusion injury
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DOI：
10.1016/s0003-4975(99)00164-2
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发表时间：
1999-05-01
影响因子：
4.6
通讯作者：
Patterson, GA
中科院分区：
文献类型：
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作者：
Hiratsuka, M;Mora, BN;Patterson, GA
背景我们最近证明，热应激诱导热休克蛋白70（HSP 70）在供体动物收获前减少保存大鼠肺移植后缺血再灌注损伤。本研究旨在探讨腺病毒介导的HSP 70基因转染大鼠同种肺移植物的可行性，并研究其表达对肺缺血再灌注损伤的影响。在确定最佳滴度的初步研究中，给动物注射不同滴度的腺病毒-HSP 70（盐水，5 × 10（9），1 × 10（10）和2 × 10（10）空斑形成单位[pfu]），并在注射后5天处死动物。为了确定最佳暴露时间，在静脉内注射腺病毒-HSP 70后的不同时间（0、6、24和72小时）处死动物。在随后的一系列移植实验中，根据转染策略将供体分配到三组。第1组（n = 8）供体静脉内接受5 × 10（9）pfu腺病毒-HSP 70，第2组（n = 7）供体接受5 × 10（9）pfu腺病毒-β-半乳糖苷酶（作为病毒对照），第3组（n = 7）供体接受盐水并作为阴性对照。处理后24小时，收集所有移植物并在原位左肺移植前储存18小时。植入后24小时，处死动物进行评估。Western blot检测HSP 70的表达。在初步研究中，即使在低滴度（5 × 10（9）pfu）的腺病毒-HSP 70也可检测到HSP 70，并且在静脉注射后6小时就可检测到低水平的HSP 70。热休克蛋白70在24小时表达最高。在移植实验中，Western印迹分析表明，HSP 70的过度表达发生在HSP 70转染的肺。再灌注后24小时平均动脉氧合上级组优于其他组（p < 0.05）。与其他组相比，第1组移植物的湿干重比（p <0.05）和髓过氧化物酶活性（p < 0.05）也显著降低。本研究表明，在体内，供体腺病毒介导的HSP 70基因转移减少随后的大鼠肺移植缺血再灌注损伤。(Ann Thorac Surg 1999;67：1421-7）（C）1999年，胸外科医师协会。
Background. We recently demonstrated that heat stress induction of heat shock protein 70 (HSP70) in donor animals before harvest decreases posttransplant ischemia-reperfusion injury in preserved rat lung isografts. The purpose of this study was to investigate the feasibility of HSP70 gene transfection into rat lung isografts using an adenoviral vector, and to study the effects of gene expression on subsequent ischemia-reperfusion injury.Methods. In preliminary studies to determine the optimal titer, animals were injected with various titers of adenovirus-HSP70 (saline, 5 x 10(9), 1 x 10(10), and 2 x 10(10) plaque forming units [pfu]) and sacrificed 5 days after injection. To determine the optimal exposure time, animals were sacrificed at different times (0, 6, 24, and 72 hours) after intravenous injection of adenovirus-HSP70. In a subsequent series of transplant experiments, donors were allocated to three groups according to transfection strategy. Group 1 (n = 8) donors received 5 x 10(9) pfu adenovirus-HSP70 intravenously, group 2 (n = 7) donors received 5 x 10(9) pfu adenovirus-beta-galactosidase (as a virus control), and group 3 (n = 7) donors received saline and served as a negative control. Twenty-four hours after treatment all grafts were harvested and stored for 18 hours before orthotopic left lung transplantation. Twenty-four hours after implantation animals were sacrificed for assessment. The expression of HSP70 was assessed by Western blot analysis.Results. In preliminary studies, HSP70 was detectable even at low titers (5 x 10(9) pfu) of adenovirus-HSP70, and was detectable at low levels as early as 6 hours after intravenous administration. Heat shock protein 70 expression was maximal at 24 hours. In transplant experiments, Western blot analysis showed that overexpression of HSP70 occurred in the HSP70-transfected lungs. The mean arterial oxygenation 24 hours after reperfusion in group 1 was superior in comparison with other groups (p < 0.05). Wet to dry weight ratio (p < 0.05) and myeloperoxidase activity (p < 0.05) were also significantly less in group 1 grafts compared with the other groups.Conclusions. This study demonstrates that in vivo, donor adenovirus-mediated gene transfer of HSP70 decreases subsequent ischemia-reperfusion injury in rat lung isografts. (Ann Thorac Surg 1999;67:1421-7) (C) 1999 by The Society of Thoracic Surgeons.