LUNG ENDOTHELIAL CELL APOPTOSIS DURING ISCHEMIC ACUTE KIDNEY INJURY
LUNG ENDOTHELIAL CELL APOPTOSIS DURING ISCHEMIC ACUTE KIDNEY INJURY
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DOI：
10.1097/shk.0b013e31826359d0
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发表时间：
2012-09-01
期刊：
影响因子：
3.1
通讯作者：
Hassoun, Heitham T.
中科院分区：
文献类型：
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作者：
White, Laura E.;Cui, Yan;Hassoun, Heitham T.
肾缺血再灌注损伤（IRI）激活细胞和可溶性介质，这些介质驱动肺部炎症级联反应、肿瘤坏死因子受体1（TNFR 1）介导的程序性细胞死亡和微血管屏障功能障碍，导致急性肺损伤。我们假设，肺微血管内皮细胞（EC），其在维持肺半透性屏障中的不可或缺的作用，是缺血性阿基期间TNFR 1介导的细胞凋亡的关键细胞靶点。雄性C57/BL 6小鼠和Sprague-Dawley大鼠进行60分钟的双侧肾蒂闭塞（IRI）或假剖腹手术（sham），并在4或24 h处死。用TUNEL、DAPI和CD 34进行共定位以鉴定EC特异性凋亡。采用新型组织消化技术和磁珠分选法分离小鼠ECs（CD 45（-）/CD 31（+）），并对84个凋亡相关基因进行实时定量聚合酶链反应Super Array分析。平行地，用来自假手术或肾IRI后获得的大鼠的血清处理生长至汇合的大鼠肺微血管EC。大鼠肺微血管内皮细胞处理+/-依那西普，TNF-α/TNFR 1信号传导抑制剂，进行定制的实时聚合酶链反应分析的促凋亡和TNF超家族转录事件，并确定凋亡与caspase 3和聚（ADP-核糖）聚合酶活性测定。在体内，TUNEL阳性细胞与CD 34共定位在全肺组织和分离的肺EC中，在缺血性阿基期间表现出促凋亡转录组。在体外，与假手术组相比，缺血性阿基在24 h时引发促凋亡（FasL、Dapk 1、Bcl 10）和TNF超家族（TNFR 1、TNFR 2、TNF-α）基因激活，并增加caspase 3和聚（ADPribose）聚合酶活性。与溶剂相比，依那西普处理大鼠肺微血管内皮细胞可抑制缺血性阿基期间促炎基因激活（E-选择素、细胞间粘附分子1、白细胞介素6、RhoB）和细胞凋亡。缺血性阿基在肺EC转录组中驱动不同的促炎和促凋亡变化，伴有TNFR 1依赖性caspase激活和程序性细胞死亡。对阿基期间肾-肺串扰的潜在EC机制的进一步研究可能会确定这种致命疾病的潜在治疗靶点。
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 Super Array 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-alpha/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-alpha) gene activation and increased caspase 3 and poly(ADPribose) 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.