Williams Rebecca E, Jacobsen Matthew, Lock Edward A
Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, SK10 4TJ, United Kingdom.
Chem Res Toxicol. 2003 Oct;16(10):1207-16. doi: 10.1021/tx030019q.
Proton NMR spectroscopy of urine has previously been used to gain insight into the site and mechanism of toxic injury to the kidney. d-Serine injures the rat kidney, causing selective necrosis of the proximal straight tubules. Damage is accompanied by proteinuria, glucosuria, and amino aciduria, the latter preceding the onset of necrosis. This study has employed (1)H NMR spectroscopy of urine and (1)H NMR and (31)P NMR spectroscopy of kidney extracts to examine the nephrotoxic action of d-serine. Urine was collected 0-8 h (all doses) and 8-24, 24-48, 48-72, 72-96, and 96-120 h (500 mg/kg only) postdosing from Alderley Park rats given d-serine (62.5, 125, 250, and 500 mg/kg ip). (1)H NMR spectra were monitored for markers of tubular damage. Additionally, ATP and ADP were quantitated in kidney perchloric acid extracts, prepared after 0.5, 1, 2, 4, and 8 h (500 mg/kg) to assess energy status; serine was also measured in these samples. At 500 mg/kg, glucosuria, amino aciduria, and reduced citrate, alpha-ketoglutarate, and succinate were observed in urine at 0-8 h. Furthermore, serine and pyruvate levels were elevated at this time. After 8-24 h, similar changes were observed; however, they were more severe reflecting the development of the lesion prior to recovery. These perturbations were dose-related, in particular, for serine and pyruvate, with no alterations seen at 62.5 mg/kg. Kidney serine concentration rapidly increased, where it was maximal after 30 min and cleared by 8 h. A decline in ATP, to approximately 60-70% of control, was observed within the kidney at 2-4 h postdosing, when necrosis first becomes evident suggesting that mitochondrial function might be impaired in the early stages of d-serine-induced nephrotoxicity. The use of NMR spectroscopy has given a comprehensive overview of the effects of d-serine in vivo. Information on the excretion of serine and its effect on renal energy metabolism provides insight into the possible mechanism of renal tubule injury.
尿液的质子核磁共振光谱先前已被用于深入了解肾脏毒性损伤的部位和机制。d-丝氨酸会损伤大鼠肾脏，导致近端直小管发生选择性坏死。损伤伴有蛋白尿、糖尿和氨基酸尿，后者在坏死开始之前就已出现。本研究采用尿液的¹H核磁共振光谱以及肾脏提取物的¹H核磁共振和³¹P核磁共振光谱来研究d-丝氨酸的肾毒性作用。给奥尔德利公园大鼠腹腔注射d-丝氨酸（62.5、125、250和500 mg/kg）后，在给药后0 - 8小时（所有剂量组）以及8 - 24、24 - 48、48 - 72、72 - 96和96 - 120小时（仅500 mg/kg剂量组）收集尿液。监测¹H核磁共振光谱以检测肾小管损伤的标志物。此外，在给药后0.5、1、2、4和8小时（500 mg/kg剂量组）制备肾脏高氯酸提取物，对其中的ATP和ADP进行定量以评估能量状态；同时也测量这些样品中的丝氨酸含量。在500 mg/kg剂量下，给药后0 - 8小时尿液中出现糖尿、氨基酸尿，以及柠檬酸盐、α-酮戊二酸和琥珀酸盐减少。此外，此时丝氨酸和丙酮酸水平升高。在8 - 24小时后，观察到类似变化；然而，这些变化更为严重，反映出损伤在恢复之前的发展情况。这些扰动与剂量相关，特别是对于丝氨酸和丙酮酸，在62.5 mg/kg剂量下未见改变。肾脏丝氨酸浓度迅速升高，在30分钟时达到最高，8小时后清除。给药后2 - 4小时，肾脏内ATP下降至对照水平的约60 - 70%，此时坏死首次明显出现，这表明在d-丝氨酸诱导的肾毒性早期阶段线粒体功能可能受损。核磁共振光谱的应用全面概述了d-丝氨酸在体内的作用。关于丝氨酸排泄及其对肾脏能量代谢影响的信息为肾小管损伤的可能机制提供了深入了解。
