Endostatin expression in the murine model of ischaemia/reperfusion-induced acute renal failure
Endostatin expression in the murine model of ischaemia/reperfusion-induced acute renal failure
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DOI：
10.1111/j.1440-1797.2007.00850.x
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发表时间：
2007-10-01
期刊：
影响因子：
2.5
通讯作者：
Schor, Nestor
中科院分区：
文献类型：
--
作者：
Bellini, Maria H.;Coutinho, Enia L.;Schor, Nestor
背景肾缺血缺氧是急性肾衰竭的主要原因，急性肾衰竭是一种与肾功能快速丧失和高死亡率相关的临床病症。肾近端小管细胞是肾缺血期间损伤最严重的细胞，由肾小管基底膜的细胞外基质的破坏引起。内皮抑制素是通过蛋白水解切割产生的胶原XVIII的C-末端片段，并且众所周知它是血管生成的抑制剂。在体外，内皮抑制素抑制内皮细胞增殖和迁移，以及小管形成。在体内，它对肿瘤生长具有有效的抑制作用。在这项研究中，我们分析了内皮抑素基因表达在C57 BL/6小鼠肾脏缺血/reperfusion.Methods：缺血性肾功能衰竭诱导通过45分钟的双侧闭塞的肾动脉和静脉，随后12小时或24小时的再灌注。提取全肾组织匀浆和总RNA，采用Western印迹分析和定量聚合酶链反应进行检测。免疫组织化学检测显示内皮抑素在损伤肾脏中表达增加，主要在近端小管和集合管。结果：内皮抑素/胶原XVIII mRNA和蛋白表达在缺血期间和再灌注12 h内增加。在蛋白质印迹分析中，我们确定了30 kDa内皮抑制素相关片段和基质金属蛋白酶-9的表达增加。CD 31在再灌注期表达显著增加（P < 0.05）。免疫组织化学检查显示肾小球和肾小管间质表达endostatin.Conclusion：这些数据表明，局部合成的30 kDa endostatin-related片段急性肾功能衰竭后，并建议其在肾毛细血管密度的调制作用。
Background. Renal ischaemia-hypoxia is a leading cause of acute renal failure, a clinical condition associated with rapid loss of renal function and high rates of mortality. Renal proximal tubular cells are the most severely injured during renal ischaemia, caused by the breakdown of the extracellular matrix of the tubular basement membrane. Endostatin is the C-terminal fragment of collagen XVIII generated by proteolytic cleavage and it is well-known as being an inhibitor of angiogenesis. In vitro, endostatin inhibits endothelial cell proliferation and migration, as well as tubule formation. In vivo, it has a potent inhibitory effect on tumour growth. In this study, we analysed endostatin gene expression in C57BL/6 mouse kidneys subjected to ischaemia/reperfusion.Methods: Ischaemic renal failure was induced via 45 min of bilateral occlusion of the renal artery and vein, followed by 12 h or 24 h of reperfusion. Whole-kidney homogenate and total RNA were extracted for examination by western blot analysis and quantitative polymerase chain reaction. The immunohistological examination revealed increased endostatin expression in injured kidney, mainly in the proximal tubule and collecting ducts.Results: Endostatin/collagen XVIII mRNA and protein expression increased during ischaemia and within 12 h of reperfusion. In the western blot assay, we identified increased expression of the 30 kDa endostatin-related fragment and of matrix metalloproteinase-9. CD31 was significantly expressed during reperfusion (P < 0.05). Immunohistological examination revealed glomerular and tubulointerstitial expression of endostatin.Conclusion: These data suggest the local synthesis of a 30 kDa endostatin-related fragment following acute renal failure and suggest its role in the modulation of renal capillary density.