Wang X, Liu K-C, Sun G-J, Han L-W, Wang R-C, Peng W-B, Sun C, Hsiao C-D, Zhang Y, Hou H-R
Biology Institute of Shandong Academy of Sciences, Jinan, China.
Biology Institute of Shandong Academy of Sciences, Jinan, China
Hum Exp Toxicol. 2016 Sep;35(9):974-82. doi: 10.1177/0960327115613844. Epub 2015 Nov 26.
To analyze the toxic effects of aristolochic acid (AA) on developed kidneys in zebrafish larvae, zebrafish at 3 days postfertilization were treated with various concentrations of AA for 24 h before the status of kidney injury was investigated from several points of view. It was found that 21% of the larvae treated with 10 µmoL/L AA exhibited evident periocular edema. When the concentrations of AA were increased to 20 and 40 µmoL/L, defect in the cardiovascular system characterized by slow heart beat and blood flow was seen coupled with periocular edema. Creatinine in the whole larval tissue determined by liquid chromatography-mass spectrometry/mass spectrometry exhibited dramatic increase in the treated groups in a dose-dependent manner within a certain range of doses. Several evident protein bands were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in supernatant of the treated larvae, indicating leakage of glomerular filtration barrier. Results of quantitative polymerase chain reaction show that the messenger RNA expression of nephrin in the 20 and 40 µmoL/L AA-treated groups decreased to 0.58 ± 0.062 and 0.37 ± 0.075-folds of the control, respectively. Kidney damage was further confirmed by the histological changes in paraffin sections of treated larvae, for example, cystic glomeruli and disorganized epithelia cells of pronephric tubules. Our results revealed that AA exerted toxic effects on developed kidney of zebrafish larvae in a dose-dependent manner and podocyte dysfunction may be involved in the kidney injury and proteinuria.
为了分析马兜铃酸（AA）对斑马鱼幼体发育中肾脏的毒性作用，在受精后3天的斑马鱼用不同浓度的AA处理24小时，然后从多个角度研究肾脏损伤情况。结果发现，用10µmoL/L AA处理的幼体中有21%出现明显的眼周水肿。当AA浓度增加到20和40µmoL/L时，除眼周水肿外，还出现了以心跳和血流缓慢为特征的心血管系统缺陷。通过液相色谱 - 质谱联用/质谱法测定的整个幼体组织中的肌酐在一定剂量范围内，处理组呈现出剂量依赖性的显著增加。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳在处理后的幼体上清液中检测到几条明显的蛋白条带，表明肾小球滤过屏障的渗漏。定量聚合酶链反应结果显示，在20和40µmoL/L AA处理组中，nephrin的信使核糖核酸表达分别降至对照组的0.58±0.062倍和0.37±0.075倍。处理后幼体石蜡切片的组织学变化进一步证实了肾脏损伤，例如囊性肾小球和前肾小管上皮细胞紊乱。我们的结果表明，AA对斑马鱼幼体发育中的肾脏具有剂量依赖性的毒性作用，足细胞功能障碍可能与肾脏损伤和蛋白尿有关。
