Yang Zequan, Sharma Ashish K, Marshall Melissa, Kron Irving L, Laubach Victor E
Department of Surgery, University of Virginia Health System, P.O. Box 801359, Charlottesville, VA 22908, USA.
Am J Respir Cell Mol Biol. 2009 Mar;40(3):375-81. doi: 10.1165/rcmb.2008-0300OC. Epub 2008 Sep 11.
Reactive oxygen species (ROS) play a crucial role in ischemia-reperfusion (IR) injury after lung transplantation. We hypothesized that NADPH oxidase derived from bone marrow (BM) cells contributes importantly to lung IR injury. An in vivo mouse model of lung IR injury was employed. Wild-type C57BL/6 (WT) mice, p47(phox) knockout (p47(phox)-/-) mice, or chimeras created by BM transplantation between WT and p47(phox)-/- mice were assigned to either Sham (left thoracotomy) or six study groups that underwent IR (1 h left hilar occlusion and 2 h reperfusion). After reperfusion, pulmonary function was assessed using an isolated, buffer-perfused lung system. Lung injury was assessed by measuring vascular permeability (via Evans blue dye), edema, neutrophil infiltration (via myeloperoxidase [MPO]), lipid peroxidation (via malondialdyhyde [MDA]), and expression of proinflammatory cytokines. Lung IR resulted in significantly increased MDA in WT mice, indicative of oxidative stress. WT mice treated with apocynin (an NADPH oxidase inhibitor) and p47(phox)-/- mice displayed significantly reduced pulmonary dysfunction and injury (vascular permeability, edema, MPO, and MDA). In BM chimeras, significantly reduced pulmonary dysfunction and injury occurred after IR in p47(phox)-/--->WT chimeras (donor-->recipient) but not WT-->p47(phox)-/- chimeras. Induction of TNF-alpha, IL-17, IL-6, RANTES (CCL5), KC (CXCL1), MIP-2 (CXCL2), and MCP-1 (CCL2) was significantly reduced after IR in NADPH oxidase-deficient mice and p47(phox)-/--->WT chimeras but not WT-->p47(phox)-/- chimeras. These results indicate that NADPH oxidase-generated ROS specifically from BM-derived cells contributes importantly to lung IR injury. NADPH oxidase may represent a novel therapeutic target for the treatment of IR injury after lung transplantation.
活性氧（ROS）在肺移植后的缺血再灌注（IR）损伤中起关键作用。我们假设源自骨髓（BM）细胞的NADPH氧化酶对肺IR损伤有重要贡献。采用了小鼠肺IR损伤的体内模型。将野生型C57BL/6（WT）小鼠、p47（phox）基因敲除（p47（phox）-/-）小鼠或通过WT和p47（phox）-/-小鼠之间的骨髓移植产生的嵌合体小鼠分为假手术组（左胸切开术）或六个接受IR的研究组（左肺门阻断1小时和再灌注2小时）。再灌注后，使用离体缓冲灌注肺系统评估肺功能。通过测量血管通透性（通过伊文思蓝染料）、水肿、中性粒细胞浸润（通过髓过氧化物酶[MPO]）、脂质过氧化（通过丙二醛[MDA]）和促炎细胞因子的表达来评估肺损伤。肺IR导致WT小鼠中MDA显著增加，表明存在氧化应激。用阿朴吗啡（一种NADPH氧化酶抑制剂）处理的WT小鼠和p47（phox）-/-小鼠表现出肺功能障碍和损伤（血管通透性、水肿、MPO和MDA）显著降低。在骨髓嵌合体中，p47（phox）-/-→WT嵌合体（供体→受体）在IR后肺功能障碍和损伤显著降低，但WT→p47（phox）-/-嵌合体则不然。在NADPH氧化酶缺陷小鼠和p47（phox）-/-→WT嵌合体中，IR后TNF-α、IL-17、IL-6、RANTES（CCL5）、KC（CXCL1）、MIP-2（CXCL2）和MCP-1（CCL2）的诱导显著降低，但WT→p47（phox）-/-嵌合体则不然。这些结果表明，特别是源自BM细胞的NADPH氧化酶产生的ROS对肺IR损伤有重要贡献。NADPH氧化酶可能是治疗肺移植后IR损伤的新治疗靶点。
