Alterations in the renal elastin-elastase system in type 1 diabetic nephropathy identified by proteomic analysis
Alterations in the renal elastin-elastase system in type 1 diabetic nephropathy identified by proteomic analysis
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DOI：
10.1097/01.asn.0000115334.65095.9b
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发表时间：
2004-03-01
影响因子：
13.6
通讯作者：
Klein, JB
中科院分区：
文献类型：
--
作者：
Thongboonkerd, V;Barati, MT;Klein, JB
糖尿病现在占终末期肾病患者的40%。尽管对糖尿病肾病的认识有了很大的进步，但导致糖尿病肾损害的细胞机制还不完全清楚。为了明确伴随糖尿病肾病的蛋白质表达的变化，将120d龄患有低胰岛素血症、高溶血症、高脂血症和蛋白尿的OVE26转基因小鼠的肾脏蛋白质组与背景FVB非糖尿病小鼠(n=5)进行了比较。从全肾裂解液中提取的蛋白质用双向PAGE分离，并用基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱仪进行鉴定。在300个可视化蛋白质点中，有41个蛋白质在糖尿病肾脏中差异表达。在这些变化的蛋白质中，单核细胞/中性粒细胞弹性蛋白酶抑制物表达增加，而弹性酶IIIB表达降低，这导致了弹性蛋白在糖尿病肾脏中表达增加的假设。325日龄FVB和OVE26小鼠的肾脏弹性蛋白免疫组织化学显示，糖尿病小鼠肾脏的致密黄斑、集合管和盆腔上皮中有显著的弹性蛋白积聚。来自1型糖尿病患者(n-3)的人肾活检组织的弹性蛋白免疫组织化学显示，弹性蛋白在肾小管细胞和间质中表达增加，但在肾小球中没有表达。这些结果表明，弹性酶抑制物和弹性酶表达的协同变化导致糖尿病肾病中弹性蛋白在肾小管间质的沉积增加。这些蛋白表达的协调变化在糖尿病肾病中的鉴定表明，蛋白质组学分析在确定病理生理学方面具有潜在的价值。
Diabetes now accounts for > 40% of patients with ESRD. Despite significant progress in understanding diabetic nephropathy, the cellular mechanisms that lead to diabetes-induced renal damage are incompletely defined. For defining changes in protein expression that accompany diabetic nephropathy, the renal proteome of 120-d-old OVE26 transgenic mice with hypoinsulinemia, hyperolycemia, hyperlipidemia, and proteinuria were compared with those of background FVB nondiabetic mice (n = 5). Proteins derived from whole-kidney lysate were separated by two-dimensional PAGE and identified by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. Forty-one proteins from 300 Visualized protein spots were differentially expressed in diabetic kidneys. Among these altered proteins, expression of monocyte/neutrophil elastase inhibitor was increased, whereas elastase IIIB was decreased, leading to the hypothesis that elastin expression would be increased in diabetic kidneys. Renal immunohistochemistry for elastin of 325-d-old FVB and OVE26 mice demonstrated marked accumulation of elastin in the macula densa, collecting ducts, and pelvicalyceal epithelia of diabetic kidneys. Elastin immunohistochemistry of human renal biopsies from patients with type 1 diabetes (n - 3) showed increased elastin expression in renal tubular cells and the interstitium but not glomeruli. These results suggest that coordinated changes in elastase inhibitor and elastase expression result in increased tubulointerstitial deposition of elastin in diabetic nephropathy. The identification of these coordinated changes in protein expression in diabetic nephropathy indicates the potential value of proteomic analysis in defining pathophysiology.