Preservation of Microvascular Integrity in Murine Orthotopic Tracheal Allografts by Clopidogrel
Preservation of Microvascular Integrity in Murine Orthotopic Tracheal Allografts by Clopidogrel
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DOI：
10.1097/tp.0000000000002571
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发表时间：
2019-05-01
期刊：
影响因子：
6.2
通讯作者：
Ensminger, Stephan M.
中科院分区：
文献类型：
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作者：
Heim, Christian;Khan, Mohammad Afzal;Ensminger, Stephan M.
背景。肺移植后的生存主要受到慢性同种异体肺移植功能障碍（CLAD）的发展的限制。本研究的目的是调查氯吡格雷抑制血小板是否对原位气管同种异体移植物的微血管完整性具有功能相关的影响，作为 CLAD 发展的解剖学基础。方法。我们将 C57Bl/6 (H-2(b)) 气管原位移植到 CBA 中。 J (H-2(k)) 接受者随后接受氯吡格雷 (1 mg/kg)。通过测量与固有层厚度成比例的上皮高度（上皮-固有层比率）来进行形态计量分析。使用荧光猝灭技术测定组织氧合，并通过激光多普勒血流计和凝集素结合测定进行移植物灌注监测。免疫组织化学检测CD31和诱导型一氧化氮合酶，普鲁士蓝反应显示铁沉积。定量逆转录聚合酶链式反应分析用于基因表达分析。结果。在整个实验过程中，同种移植物保持良好的氧合和灌注，而同种异体移植物中两者均急剧减少。氯吡格雷治疗可减轻移植物缺氧并减少灌注损失。此外，氯吡格雷导致上皮-固有层比率增加，同时铁沉积受损。基因表达分析显示氯吡格雷组的血管生成血管内皮生长因子水平升高。免疫组织化学（CD31，诱导型一氧化氮合酶）显示内皮功能得到改善。结论。持续给予氯吡格雷可显着改善组织氧合，限制微血管渗漏，并预防气道缺血。这些数据表明，氯吡格雷可改善急性气道排斥反应期间的微血管损伤，这是发生 CLAD 的已知诱发因素。
Background. Survival after lung transplantation is mainly limited by the development of chronic lung allograft dysfunction (CLAD). The aim of this study was to investigate if platelet inhibition by clopidogrel has a functionally relevant influence on the microvascular integrity of orthotopic tracheal allografts as an anatomic basis for the development of CLAD. Methods. We orthotopically transplanted C57Bl/6 (H-2(b)) tracheas into CBA. J (H-2(k)) recipients who afterwards received clopidogrel (1 mg/kg). Morphometric analysis was performed by measuring epithelial height in proportion to thickness of the lamina propria (epithelium-lamina propria ratio). Tissue oxygenation was determined using a fluorescence quenching technique, and graft perfusion monitoring was performed by laser Doppler flowmetry and lectin-binding assay. Immunohistochemistry was used for detection of CD31 and inducible nitric oxide synthase while iron deposition was shown with Prussian blue reaction. Quantitative reverse transcription polymerase chain reaction analysis was used for gene expression analysis. Results. Isografts maintained good oxygenation and perfusion throughout the experiment, while both were drastically reduced in allografts. Treatment with clopidogrel attenuated graft hypoxia and reduced loss of perfusion. Additionally, clopidogrel led to increased epithelium-lamina propria ratio while iron deposition was impaired. Gene expression analysis revealed elevated levels of angiogenic vascular endothelial growth factor in the clopidogrel group. Improved endothelial function was shown by immunohistochemistry (CD31, inducible nitric oxide synthase). Conclusions. Continuous administration of clopidogrel significantly improved tissue oxygenation, limited microvascular leakiness, and prevented airway ischemia. These data demonstrate that clopidogrel ameliorates microvascular injury during acute airway rejection, which is a known predisposing factor for the development of CLAD.