School of Pharmacology, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Acta Pharmacol Sin. 2011 Jul;32(7):888-94. doi: 10.1038/aps.2011.39. Epub 2011 Jun 20.
To identify the key proteins involved in the nephrotoxicity induced by andrographolide sodium bisulfite (ASB).
Male ICR mice were intravenously administrated with ASB (1000 or 150 mg·kg⁻¹·d⁻¹) for 7 d. The level of malondialdehyde (MDA) and the specific activity of superoxide dismutase (SOD) in kidneys were measured. The renal homogenates were separated by two-dimensional electrophoresis, and the differential protein spots were identified using a matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF)/TOF mass spectrometry.
The high dose (1000 mg/kg) of ASB significantly increased the MDA content, but decreased the SOD activity as compared to the control mice. The proteomic analysis revealed that 6 proteins were differentially expressed in the high-dose group. Two stress-responsive proteins, ie heat shock cognate 71 kDa protein (HSC70) and peroxiredoxin-6 (PRDX6), were regulated at the expression level. The remaining 4 proteins involving in cellular energy metabolism, including isoforms of methylmalonyl-coenzyme A mutase (MUT), nucleoside diphosphate-linked moiety X motif 19 (Nudix motif19), mitochondrial NADH dehydrogenase 1 alpha subcomplex subunit 10 (NDUFA10) and nucleoside diphosphate kinase B (NDK B), were modified at the post-translational levels.
Our findings suggest that the mitochondrion is the primary target of ASB and that ASB-induced nephrotoxicity results from oxidative stress mediated by superoxide produced by complex I.
鉴定与穿心莲内酯亚硫酸氢钠（ASB）诱导的肾毒性相关的关键蛋白。
雄性 ICR 小鼠静脉注射 ASB（1000 或 150mg·kg⁻¹·d⁻¹）7d。测定肾脏丙二醛（MDA）水平和超氧化物歧化酶（SOD）比活性。二维电泳分离肾匀浆，基质辅助激光解吸/电离（MALDI）飞行时间/飞行时间（TOF）质谱鉴定差异蛋白点。
与对照组相比，高剂量（1000mg/kg）ASB 显著增加 MDA 含量，但降低 SOD 活性。蛋白质组学分析显示，高剂量组有 6 种蛋白表达差异。两种应激反应蛋白，即热休克同源 71kDa 蛋白（HSC70）和过氧化物酶 6（PRDX6），在表达水平上受到调节。其余 4 种涉及细胞能量代谢的蛋白，包括甲基丙二酰辅酶 A 变位酶（MUT）同工酶、核苷二磷酸连接部分 X 基序 19（Nudix 基序 19）、线粒体 NADH 脱氢酶 1α亚基 10（NDUFA10）和核苷二磷酸激酶 B（NDK B），在翻译后水平上发生改变。
我们的研究结果表明，线粒体是 ASB 的主要靶标，ASB 诱导的肾毒性是由复合体 I 产生的超氧阴离子介导的氧化应激引起的。
