Identification and kinetics of leukocytes after severe ischaemia/reperfusion renal injury

作者：Dirk Ysebaert, Kathleen E. De Greef, Sven R. Vercauteren, Manuela Ghielli, Gert A. Verpooten, E Eyskens, Marc E. De Broe · 发表于：Nephrology Dialysis Transplantation · 年份：2000 · DOI：10.1093/ndt/15.10.1562 · 被引用次数：375 · 研究领域：Neutrophil, Myeloperoxidase and Oxidative Mechanisms、Immune Response and Inflammation、Neuroinflammation and Neurodegeneration Mechanisms

BACKGROUND: Leukocyte adhesion/infiltration in response to renal ischaemia/reperfusion (I/R) injury is a well-known but poorly understood phenomenon. The identification, kinetics, and exact role of these inflammatory cells in I/R injury and regeneration are still matters of debate. METHODS: Uninephrectomized rats were submitted to 60 min renal ischaemia by clamping of renal vessels. RESULTS: Severe acute renal failure was observed, with maximum functional impairment on day 2. By 12 h after the ischaemic event, up to 80% of proximal tubular cells in the outer stripe of outer medulla (OSOM) were already severely damaged. Proliferation (proliferating cell nuclear antigen (PCNA) staining) started after 24 h, reaching maximum activity on day 3. Regeneration of tubular morphology started on the 3rd day, and after 10 days 50% of tubules had regenerated completely. Interstitial leukocytes (OX-1 immunohistochemical staining) were already prominent at day 1, thereafter gradually increasing with time. The so-called neutrophil-specific identification methods (myeloperoxidase (MPO), chloroacetate esterase, mAb HIS-48) proved to be non-specific, since they also stained for macrophages, as demonstrated by flow cytometry and the combination of these stainings with the macrophage-specific ED-1 staining. MPO activity was already significantly increased at 1 h post-I/R (439+/-34%, P<0.005), reaching its maximum activity after 12 h of I/R (1159+/-138%, P<0.0005), declining thereafter. On the oth...