Localization of Aristolochic Acid in Mouse Kidney Tissues by Immunohistochemistry Using an Anti-AA-I and AA-II Monoclonal Antibody
Localization of Aristolochic Acid in Mouse Kidney Tissues by Immunohistochemistry Using an Anti-AA-I and AA-II Monoclonal Antibody
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DOI：
10.1142/s0192415x14500918
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发表时间：
2014-01-01
影响因子：
5.7
通讯作者：
Cai, Shao-Qing
中科院分区：
文献类型：
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作者：
Li, Xiao-Wei;Yokota, Sadaki;Cai, Shao-Qing
马兜铃酸（Aristolochic Acids，AA）存在于马兜铃科植物（包括马兜铃属和细辛属）的草药中。AAs与快速进展的间质性肾炎相关，称为马兜铃酸肾病（AAN）。然而，在原位本地化的AA在靶器官，肾脏，尚未进行研究。本研究采用免疫过氧化物酶光镜和胶体金免疫电镜（IEM）技术，观察了马兜铃酸Ⅰ（AA-Ⅰ）和AA-Ⅱ单克隆抗体（mAb）在小鼠肾脏中的蓄积。雄性BALB/c小鼠每天给予1.25或2.50 mg/kg AA-1，持续5天。分别制备肾组织石蜡切片和超薄切片。在光学显微镜下，近端小管的顶端表面强烈的AA-I染色，而没有明显的免疫染色被发现在远端小管和肾小球，保持相对完整。电镜下可见近曲小管、远曲小管和集合小管上皮细胞不同程度的断裂。近曲小管和远曲小管的金标记较集合小管强。在肾小管，AA-Ⅰ的免疫金信号倾向于积累在线粒体和过氧化物酶体，虽然信号可以观察到整个细胞。在肾小球的红细胞中也发现了金信号。抗AA-I和AA-II单克隆抗体的获得为寻找与AA-I相互作用的蛋白质或因子，从而诱导肾靶向损伤提供了线索。
Aristolochic acids (AAs) are found in herbal medicines of Aristolochiaceae plants, including Aristolochia and Asarum species. AAs are associated with a rapidly progressive interstitial nephritis, which is called aristolochic acid nephropathy (AAN). However, the in-situ localization of AAs in the target organ, the kidney, has not been investigated yet. In the present study, the accumulation of aristolochic acid I (AA-I) in mouse kidney was revealed by immunoperoxidase light microscopy as well as colloidal gold immunoelectron microscopy (IEM) based on an anti-AA-I and AA-II monoclonal antibody (mAb). Male BALB/c mice were treated with 1.25 or 2.50 mg kg(-1) of AA-I per day for 5 days. Paraffin sections and ultra-thin sections of kidney tissue were respectively prepared. Under light microscopy, the apical surface of proximal tubules was strongly stained for AA-I, whereas no obvious immunostaining was found in the distal tubules and glomerulus, which remained relatively intact. Under electron microscopy, epithelial cells of the proximal tubules, distal tubules and collecting tubules were broken to various degrees. Gold labeling in the proximal and distal tubules was stronger than that in the collecting tubules. In renal tubules, immunogold signals of AA-I tended to accumulate in the mitochondria and peroxisomes, though the signals could be observed all over the cell. Gold signals were also found in the erythrocytes of glomeruli. The MAb against AA-I and AA-II provides a clue for the identification of proteins or factors which might interact with AA-I and thus induce targeted damage of kidney.