Metabonomics with¹H-NMR spectroscopy and liquid chromatography-mass spectrometry applied to the investigation of metabolic changes caused by gentamicin-induced nephrotoxicity in the rat

Abstract

The model nephrotoxin gentamicin was administered to male Wistar-derived rats daily, for 7 days, at 60 mg kg-1 day-1, subcutaneously, twice daily. Conventional clinical chemistry urinalysis showed a significant increase in N-acetyl-beta-D-glucosaminidase (NAG) activity from day 3. At necropsy on day 9, clear histological damage to the kidney was noted with all animals showing a generally severe nephropathy primarily focused on the proximal convoluted tubules. The urinary excretion pattern of endogenous metabolites over the time course of the study was studied using a combination of 1H-NMR spectroscopy and HPLC-TOF-MS/MS using electrospray ionization (ESI). Changes in the pattern of endogenous metabolites as a result of daily administration of gentamicin were readily detected by both techniques with significant perturbations of the urinary profile observed from day 7 onwards. The findings by 1H-NMR included raised glucose and reduced trimethylamine N-oxide (TMAO). Changes in metabonomic profiles were observed by HPLC-MS in both positive and negative ESI. The MS data showed reduced xanthurenic acid and kynurenic acid, whilst neutral loss experiments also revealed a changed pattern of sulphate conjugation on gentamicin administration.

Urinary Metabolic Phenotyping Differentiates Children with Autism from Their Unaffected Siblings and Age-Matched Controls

“…36 Depletion of hippurate and PAG has been reported after ingestion of antibiotics such as vancomycin 12 and gentamicin. 37 In addition, gastrointestinal microbial analysis of fecal samples from children with late-onset autism has shown that autistic individuals have higher incidence of the Clostridium histolyticum group of bacteria, as well as increased diversity of clostridial species 11,13 , and individuals with autism have been shown to have abnormalities in hippurate excretion. 38 Abnormalities of the gut microbiota including the Clostridium sp.…”

Metabolomics as an Extension of Proteomic Analysis: Study of Acute Kidney Injury

“…Our results confirm previous studies where quantitative changes by NMR spectroscopic metabolite patterns provided information on the location and severity of toxic lesions. Those studies detected the presence of glucosuria and aminoaciduria after exposure to known proximal tubule nephrotoxins such as gentamicin, mercuric chloride, and D-serine (7,8,48). Glucosuria and enhanced excretion of amino acids are both strong indicators of proximal tubule damage in general, likely caused by impaired tubular reabsorption in the proximal tubule via sodium-dependent glucose transport.…”

Discovery of Metabolomics Biomarkers for Early Detection of Nephrotoxicity

“…A further application was investigated by Dieterle et al (2006), who reported a metabolomics analysis in a compound ranking study, which revealed increased excretion of choline in rat urine. Several other reports have been published on the metabolomic analysis of individual nephrotoxins, such as cadmium chloride (Griffin et al 2001), mercury chloride (Holmes et al 2000), gentamicin (Lenz et al 2005), cisplatin (Portilla et al 2006), and aristolochate (Ni et al 2007), among others. Finally, a large comprehensive study was reported from the Consortium on Metabonomic Toxicology (COMET) (Ebbels et al 2007), in which many nephrotoxins were analyzed, to build prediction and classification models for drug toxicity.…”