Lung endothelial cell apoptosis during ischemic acute kidney injury
| Autor: | Heitham T. Hassoun, Laura E. White, Mihaela L. Lie, Yan Cui, Carolyn M. Feltes Shelak |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
CD31
Male Programmed cell death Acute Lung Injury Caspase 3 Apoptosis Lung injury Critical Care and Intensive Care Medicine Fas ligand Receptors Tumor Necrosis Factor Proinflammatory cytokine Etanercept Rats Sprague-Dawley Mice Medicine Animals Lung Cells Cultured business.industry Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha Anti-Inflammatory Agents Non-Steroidal Endothelial Cells Pneumonia respiratory system Acute Kidney Injury Rats Mice Inbred C57BL Receptors Tumor Necrosis Factor Type I Immunoglobulin G Reperfusion Injury Microvessels Emergency Medicine Cancer research Tumor necrosis factor receptor 1 Poly(ADP-ribose) Polymerases business Signal Transduction |
| Zdroj: | Shock (Augusta, Ga.). 38(3) |
| ISSN: | 1540-0514 |
| Popis: | Kidney ischemia-reperfusion injury (IRI) activates cellular and soluble mediators that drive lung inflammatory cascades, tumor necrosis factor receptor 1 (TNFR1)-mediated programmed cell death, and microvascular barrier dysfunction, leading to acute lung injury. We hypothesized that lung microvascular endothelial cells (ECs), with their integral role in maintaining the lung-semipermeable barrier, were key cellular targets of TNFR1-mediated apoptosis during ischemic AKI. Male C57/BL6 mice and Sprague-Dawley rats underwent 60 min of bilateral renal pedicle occlusion (IRI) or sham laparotomy (sham) and were killed at 4 or 24 h. Colocalization with TUNEL, DAPI, and CD34 was performed to identify EC-specific apoptosis. Mouse ECs (CD45/CD31) isolated with novel tissue digestion techniques and magnetic microbead sorting underwent quantitative real-time polymerase chain reaction SuperArray analysis with 84 apoptosis-related genes. In parallel, rat lung microvascular ECs grown to confluence were treated with serum from rats obtained following sham or kidney IRI. Rat lung microvascular ECs treated +/- etanercept, a TNF-α/TNFR1 signaling inhibitor, underwent custom real-time polymerase chain reaction analysis for proapoptotic and TNF superfamily transcriptional events, and apoptosis was identified with caspase 3 and poly(ADP-ribose) polymerase activity assays. In vivo, TUNEL-positive cells colocalized with CD34 in whole-lung tissue and isolated lung ECs demonstrated a proapoptotic transcriptome during ischemic AKI. In vitro, ischemic AKI incited proapoptotic (FasL, Dapk1, Bcl10) and TNF superfamily (TNFR1, TNFR2, TNF-α) gene activation and increased caspase 3 and poly(ADP-ribose) polymerase activity at 24 h versus sham. Compared with vehicle, treatment of rat lung microvascular ECs with etanercept inhibited proinflammatory gene activation (E-selectin, intercellular adhesion molecule 1, interleukin 6, RhoB) and apoptosis during ischemic AKI. Ischemic AKI drives distinct proinflammatory and proapoptotic changes in the pulmonary EC transcriptome with TNFR1-dependent caspase activation and programmed cell death. Further investigation of potential EC mechanisms of kidney-lung crosstalk during AKI may identify potential therapeutic targets for this deadly disease. |
| Databáze: | OpenAIRE |
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