INSERM U1082 (IRTOMIT), 86000 Poitiers, France.
Service de Biochimie, CHU Poitiers, 86000 Poitiers, France.
Int J Mol Sci. 2021 Nov 9;22(22):12103. doi: 10.3390/ijms222212103.
Renal ischaemia reperfusion (I/R) triggers a cascade of events including oxidative stress, apoptotic body and microparticle (MP) formation as well as an acute inflammatory process that may contribute to organ failure. Macrophages are recruited to phagocytose cell debris and MPs. The tyrosine kinase receptor MerTK is a major player in the phagocytosis process. Experimental models of renal I/R events are of major importance for identifying I/R key players and for elaborating novel therapeutical approaches. A major aim of our study was to investigate possible involvement of MerTK in renal I/R. We performed our study on both natural mutant rats for MerTK (referred to as RCS) and on wild type rats referred to as WT. I/R was established by of bilateral clamping of the renal pedicles for 30' followed by three days of reperfusion. Plasma samples were analysed for creatinine, aspartate aminotransferase (ASAT), lactate dehydrogenase (LDH), kidney injury molecule -1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) levels and for MPs. Kidney tissue damage and CD68-positive cell requirement were analysed by histochemistry. monocyte chemoattractant protein-1 (MCP-1), myeloperoxidase (MPO), inducible nitric oxide synthase (iNOS), and histone 3A (H3A) levels in kidney tissue lysates were analysed by western blotting. The phagocytic activity of blood-isolated monocytes collected from RCS or WT towards annexin-V positive bodies derived from cultured renal cell was assessed by fluorescence-activated single cell sorting (FACS) and confocal microscopy analyses. The renal I/R model for RCS rat described for the first time here paves the way for further investigations of MerTK-dependent events in renal tissue injury and repair mechanisms.
肾缺血再灌注(I/R)引发了一系列事件,包括氧化应激、凋亡小体和微颗粒(MP)的形成以及急性炎症过程,这些都可能导致器官衰竭。巨噬细胞被招募来吞噬细胞碎片和 MPs。酪氨酸激酶受体 MerTK 是吞噬过程中的主要参与者。肾 I/R 事件的实验模型对于确定 I/R 的关键因素和制定新的治疗方法非常重要。我们研究的主要目的是研究 MerTK 是否参与肾 I/R。我们在天然突变型 MerTK 大鼠(称为 RCS)和野生型大鼠(称为 WT)上进行了研究。通过双侧夹闭肾蒂 30' 来建立 I/R,然后进行三天的再灌注。分析血浆样本中的肌酐、天门冬氨酸氨基转移酶(ASAT)、乳酸脱氢酶(LDH)、肾损伤分子-1(KIM-1)和中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平以及 MPs。通过组织化学分析肾脏组织损伤和 CD68 阳性细胞的需求。通过 Western blot 分析肾组织裂解物中的单核细胞趋化蛋白-1(MCP-1)、髓过氧化物酶(MPO)、诱导型一氧化氮合酶(iNOS)和组蛋白 3A(H3A)水平。通过荧光激活的单细胞分选(FACS)和共聚焦显微镜分析,评估从 RCS 或 WT 血液中分离的单核细胞对来自培养的肾细胞的 Annexin-V 阳性体的吞噬活性。这里首次描述的 RCS 大鼠肾 I/R 模型为进一步研究 MerTK 依赖性事件在肾组织损伤和修复机制中的作用铺平了道路。