Williams R E, Major H, Lock E A, Lenz E M, Wilson I D
Department of Drug Metabolism and Pharmacokinetics, AstraZeneca, Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TG, UK.
Toxicology. 2005 Feb 14;207(2):179-90. doi: 10.1016/j.tox.2004.08.023.
HPLC-MS-based metabonomic analysis was used to investigate urinary metabolic perturbations associated with D-serine-induced nephrotoxicity. D-Serine causes selective necrosis of the proximal straight tubules in the rat kidney accompanied by aminoaciduria, proteinuria and glucosuria. Alderely Park (Wistar-derived) rats were dosed with either D-serine (250 mg/kg ip) or vehicle (deionised water) and urine was collected at 0-12, 12-24, 24-36 and 36-48 h post-dosing. Samples were analysed using a Waters Alliance HT 2795 HPLC system coupled to a Waters Micromass Q-ToF-micro equipped with an electrospray source operating in either positive or negative ion mode. Changes to the urinary profile were detected at all time points compared to control. In negative ion mode, increases were observed in serine (m/z=103.0077), m/z=104.0376 (proposed to be hydroxypyruvate) and glycerate (m/z=105.0215), the latter being metabolites of D-serine. Furthermore, an increase in tryptophan, phenylalanine and lactate and decreases in methylsuccinic acid and sebacic acid were observed. Positive ion analysis revealed a decrease in xanthurenic acid, which has previously been assigned and reported using HPLC-MS following exposure to mercuric chloride and cyclosporine A. A general aminoaciduria, including proline, methionine, leucine, tyrosine and valine was also observed as well as an increase in acetyl carnitine. Investigation of additional metabolites altered as a result of exposure to D-serine is on-going. Thus, HPLC-MS-based metabonomic analysis has provided information concerning the mechanism of D-serine-induced renal injury.
基于高效液相色谱-质谱联用的代谢组学分析用于研究与D-丝氨酸诱导的肾毒性相关的尿液代谢紊乱。D-丝氨酸可导致大鼠肾脏近端直小管选择性坏死，并伴有氨基酸尿、蛋白尿和糖尿。将奥尔德利园（源自Wistar）大鼠分为两组，分别给予D-丝氨酸（250mg/kg腹腔注射）或赋形剂（去离子水），并在给药后0-12、12-24、24-36和36-48小时收集尿液。使用与配备电喷雾源的沃特世Micromass Q-ToF-micro联用的沃特世Alliance HT 2795高效液相色谱系统对样品进行分析，该电喷雾源可在正离子或负离子模式下运行。与对照组相比，在所有时间点均检测到尿液图谱的变化。在负离子模式下，观察到丝氨酸（m/z=103.0077）、m/z=104.0376（推测为羟基丙酮酸）和甘油酸（m/z=105.0215）增加，后者是D-丝氨酸的代谢产物。此外，还观察到色氨酸、苯丙氨酸和乳酸增加，以及甲基琥珀酸和癸二酸减少。正离子分析显示，黄尿酸减少，此前在暴露于氯化汞和环孢素A后使用高效液相色谱-质谱联用对其进行了鉴定和报道。还观察到包括脯氨酸、蛋氨酸、亮氨酸、酪氨酸和缬氨酸在内的一般氨基酸尿以及乙酰肉碱增加。对因暴露于D-丝氨酸而改变的其他代谢产物的研究正在进行中。因此，基于高效液相色谱-质谱联用的代谢组学分析提供了有关D-丝氨酸诱导肾损伤机制的信息。
