Propiverine-induced accumulation of nuclear and cytosolic protein in F344 rat kidneys: Isolation and identification of the accumulating protein
Propiverine-induced accumulation of nuclear and cytosolic protein in F344 rat kidneys: Isolation and identification of the accumulating protein
复制标题
DOI：
10.1016/j.taap.2008.09.014
复制
发表时间：
2008-12-15
影响因子：
3.8
通讯作者：
Day, B. W.
中科院分区：
文献类型：
--
作者：
Dietrich, D. R.;Heussner, A. H.;Day, B. W.
治疗神经源性逼尿肌过度活动(NDO)和膀胱过度活动(OAB)患者的F344雄性和雌性F344大鼠(而不是B6C3F1)B6C3F1(NICE)暴露104周后，肾脏肿瘤的发病率没有增加，但仍未观察到肾脏近端小管上皮细胞胞浆和细胞核中蛋白质的聚集。为了更好地解释这些发现，并为人类健康风险评估提供更好的基础，F344雄性和雌性大鼠在饮食中暴露于1000ppm的普罗维林16周，通过胞浆和细胞核准备或激光捕获显微切割从肾脏分离积累的蛋白质，并使用分子量测定和质谱仪进行分析。积累的蛋白质是D-氨基酸氧化酶(DAAO)，这是一种与氨基酸和脂肪酸代谢有关的酶。随后对肾脏匀浆和细胞核样本以及组织切片进行的蛋白质印迹和DAAO免疫组织化学重新分析，证实了DAAO在普罗维林加热的雄性和雌性F344大鼠中的存在和定位。DAAO仅在大鼠体内蓄积，且大鼠DAAO与包括人类在内的其他物种的相似性有限，这表明药物诱导的肾脏DAAO蓄积是大鼠特有的机制，而与长期服用异丙维林的患者几乎没有相关性。(C)2008 Elsevier Inc.保留所有权利。
Male and female F344 rats but not B6C3F1 (nice exposed for 104 weeks to propiverine hydrochloride (1-methylpiperid-4-yl 2,2-Biphenyl-2-(1-piopoxy)acetate hydrochloride), used for treatment of patients with neurogenic detrusor overactivity (NDO) and overactive bladder(OAB), presented with an accumulation of proteins in the cytosol and nuclei of renal proximal tubule epithelial cells, yet despite this, no increased renal tumor incidence was observed. In order to provide an improved interpretation of these findings and a better basis for human health risk assessment, male and female F344 rats were exposed for 16 weeks to 1000 ppm propiverine in the diet, the accumulating protein was isolated from the kidneys via cytosolic and nuclear preparations or laser-capture microdissection and analyzed using molecular weight determination and mass spectrometry. The accumulating protein was found to be D-amino acid oxidase (DAAO), an enzyme involved in amino and fatty acid metabolism. Subsequent reanalysis of kidney homogenate and nuclear samples as well as tissue sections using western blot and DAAO-immunohistochemistry, confirmed the presence and localization of DAAO in propiverine-heated male and female F344 rats. The accumulation of DAAO only in rats, and the limited similarity of rat DAAO with other species, including humans, suggests a rat-specific mechanism underlying the drug-induced renal DAAO accumulation with little relevance for patients chronically treated with propiverine. (C) 2008 Elsevier Inc. All rights reserved.