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NMR spectroscopy and electron microscopy identification of metabolic and ultrastructural changes to the kidney following ischemia-reperfusion injury

机译:NMR光谱和电子显微镜鉴定缺血再灌注损伤后对肾脏的代谢和超微结构变化

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Cellular, molecular, and ultra-structural nephron changes associated wilh ischemia-reperfusion injury-induced acute kidney injury (IRI-AKI) are not completely understood. Here, a multidisciplinary study was used to identify nephron changes in a mouse model of IRI-AKI. Histological analyses indicated distended Bowman's glomerular spaces and proximal and distal tubules. Increased filtrate volume in nephrons was caused by reduced water reabsorption by severely damaged proximal tubule brush borders and blocked flow of filtrate into collecting tubules by mucopro-tein casts in distal tubules. Immunohistochemistry revealed protein AKI biomarkers in proximal tubules and glomeruli but not in distal tubules. Nuclear magnetic resonance spectroscopy revealed several metabolites that increased such as valine, alanine, and lactate. Other metabolites such as trigonelline, succinate, 2-oxoisocaproate, and 1-methyl-nicotinamide decreased or were absent in urine following IRI due to altered kidney function or metabolism. Urinary glucose increased due to reduced reabsorption by damaged proximal tubule brush borders. Scanning electron microscopy revealed flattening of podocytes and pedicals surrounding glomerular capillaries, and transmission electron microscopy (TEM) revealed effacement of podocyte pedicals, both consistent with increased hydrostatic pressure in nephrons following IRI-AKI. TEM revealed shortened proximal tubule microvilli in IRI kidneys with diminished lamina propia. TEM showed dramatic loss of mitochondria in distal tubule epithelia of IRI kidneys and emergence of multivesicular bodies of endosomes indicating ongoing cellular death. Collectively, the data define ultrastructural changes to nephrons and altered kidney metabolism associated with IRI-AKI.
机译:细胞,分子和超结构性肾病变化相关的威尔赫缺血再灌注损伤诱导急性肾损伤(IRI-AKI)未得到完全理解。在这里,使用多学科研究来识别IRI-AKI鼠标模型中的肾功能。组织学分析表明了鲍曼的肾小球空间和近端和远端小管。通过严重受损的近端小管刷边框减少水重吸收并通过在远端小管中通过粘液铸造粘附到收集小管中,通过减少水重吸收来增加肾脏中的滤液引起的。免疫组织化学在近端小管和肾小球中揭示了蛋白质aki生物标志物,但不在远端小管中。核磁共振光谱揭示了几种代谢物,这些代谢物如缬氨酸,丙氨酸和乳酸。由于肾功能或代谢改变,IRI后,其他代谢物如三角形,琥珀酸酯,2-氧代偶氮基,和1-甲基 - 烟酰胺降低或不存在于IRI之后。由于损伤的近端小管刷边框,尿葡萄糖增加。扫描电子显微镜显示肾小球毛细血管围绕肾小粒细胞和姿势的平坦化,透射电子显微镜(TEM)揭示了多粒细胞姿势的诱导,这两者都是一致的IRI-AKI后肾脏的静水压力增加。 TEM在IRI肾脏中透露缩短的近端小管MICROVILLI,具有减少的LAMINA PRIA。 TEM表现出在IRI肾脏的远端小管上皮细胞上的线粒体丧失,以及表明持续细胞死亡的内体的多产体的出现。统称,数据将超微结构的变化定义对肾脏和改变与IRI-AKI相关的肾脏代谢。

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