NMR-based metabonomic study on the subacute toxicity of aristolochic acid in rats
Introduction
NMR spectroscopy of biofluids has brought a new chemical method to help identify the site and possible mechanism of toxicity and offered much information of endogenous metabolites and their variation in pathological states (Nicholson and Wilson, 1989, Lindon et al., 1999, De Graaf and Behar, 2003, Constantinou et al., 2004). Numerous studies have demonstrated the utility of NMR spectroscopy in toxicological fields, especially in the study of nephrotoxins (Holmes et al., 1992, Anthony et al., 1994, Gartland et al., 1989, Nicholson et al., 1989) and hepatotoxins (Beckwith-Hall et al., 2003, Waters et al., 2001, Singh et al., 2003). It provides a method of monitoring the toxicological processes of xenobiotics and clinical toxicology (Maschke et al., 1995, Bales et al., 1988, Wu et al., 2004). With the improvements in instrument sensitivity and relative resolution of NMR spectrometers at higher magnetic field strengths which gives a greater sensitivity to the methodology increasing the amount of biochemical information from 1H NMR spectra. Pattern recognition (PR) analysis is performed in a multidimensional parameter space using dimension–reduction techniques to maximize information from NMR profile (Xu, 2004). Several methods, such as principal component analysis (PCA) and non-linear mapping (NLM) techniques (Lindon et al., 1999) have been used to classify toxins on the basis of 1H NMR spectra of urine. The application of 1H NMR spectroscopy to the study of metabolites in biofluids combined with pattern recognition to classify NMR-derived data has led to a ‘metabonomic’ approach to biochemical assessment (Holmes et al., 1998b). As a method of monitoring the progression of toxicity and recovery caused by toxins or xenobiotics, metabonomics represents a systemic approach for measuring time-related biochemical responses of metabolic composition variance (Keun et al., 2002, Lindon et al., 2003, Wu et al., 2005). Principal components analysis (PCA) is a widely used dimension–reduction method, in which the NMR spectra can be reduced to a set of peak intensity descriptors, and used to detect toxicity and disease based on NMR biomarkers as well as identify similarities or differences between the samples from control and toxin-treated animals.
Aristolochic acid (AA) is an active component derived from Aristolochia species, which is used to treat arthritis, gout and rheumatism. In addition, its anti-fungal, anti-bacterial, anti-viral, and even anti-tumor pharmacological effects have been reported (Zhang et al., 2004). However, AA was considered to be related to a progressive renal fibrosis, which was so-called ‘Chinese-herb nephropathy’ (CHN) (Vanherweghem et al., 1993). The histological findings in renal superficial cortex from ‘CHN’ patients were interstitial fibrosis and tubular atrophy (Cosyns et al., 1994, Depierreux et al., 1994). AA is metabolized into aristolactam in liver and DNA adducts have been detected in kidney and liver tissues (Debelle et al., 2003). The mutations were also detected in p53 and H-ras genes which were associated with carcinogenesis process (Arlt et al., 2000, Arlt et al., 2001). AA-induced renal fibrosis and chronic renal failure model was established in salt-depleted Wistar rats (Debelle et al., 2002). However, the subacute toxicological process caused by AA is still uncertain. As the characteristic toxicity of AA is severe, the toxic process and biochemical effect of AA should be carefully studied. In the present paper, the onset, development and recovery toxicological processes of AA to the rats were investigated by NMR-based metabonomic methods. Model toxins causing hepatic or renal lesion, which were well documented by NMR spectroscopy, were used to enable comparisons of the urinary NMR profiles to be made with ones obtained from animals treated with known toxicants.
Section snippets
Selection of the model toxins and determination of AA dose
AA was purchased from Sigma-Aldrich, sodium chromate (NaCrO4), mercury II chloride (HgCl2), 2-bromoethanamine hydrobromide (BEA), hydrazine dihydrochloride (HYD) and α-naphthylisothiocyanate (ANIT) were of molecular biology or analytical grade. NaCrO4, HgCl2 and BEA are well-known nephrotoxins, which is typically damage S1 renal tubule, S2/S3 renal tubule and the renal papillary (Holmes et al., 1998a, Gartland et al., 1989). HYD (N2H4 · 2HCl) can causes fatty liver without necrosis (Nicholls et
1H NMR analysis of urine samples from the rats treated with model toxins
A typical 600 MHz 1H NMR spectra of urine sample of 24–32 h time interval from the model and control groups were illustrated in Fig. 1. The increase of glucose and amino acids was observed in NaCrO4-treated rats from the 1H NMR spectral analysis, which agreed with the report of Holmes et al. (1998a). The slight elevation of taurine and trimethylamine N-oxide (TMAO) level was also observed. Considerable increase of the concentration of amino acids (valine and glycine) and glucose in the urine from
1H NMR spectra analysis of the urine from the rats treated with AA
Different sites and the extent of the lesion and organ damage could be associated with the corresponding changes in metabolic profiles. NaCrO4 may have damaged the proximal convoluted tubule cells (Holmes et al., 1998a), with resulting in adverse kidney function (Tandon, 1982). The 1H NMR spectral profile of urine from AA group in 48–56 h interval was similar to that of NaCrO4 group, including the increase of amino acids, glucose and taurine as well as the decrease of citrate. The results
Conflict of interest statement
There are no conflicts about this work.
Acknowledgment
This work was supported by the National Natural Science Foundation of China (Grant 20575065).
References (38)
Neprolithiasis in renal tubular acidosis
J. Urol.
(1989)- et al.
Chinese herbs nephropathy: a clue to Balkan endemic nephropathy?
Kidney Int.
(1994) - et al.
Application of nuclear magnetic resonance spectroscopy combined with principal component analysis in detecting inborn errors of metabolism using blood spots: a metabonomic approach
Anal. Chim. Acta
(2004) - et al.
Pathologic aspects of a newly described nephropathy related to the prolonged use of Chinese herbs
Am. J. Kidney Dis.
(1994) - et al.
The identification of novel biomarkers of renal toxicity using automatic data reduction techniques and PCA of proton NMR spectra of urine
Chemom. Int. Lab. Sys.
(1998) - et al.
NMR spectroscopy of biofluid
Ann. Rep. NMR. Spectrosc.
(1999) - et al.
High resolution proton magnetic resonance spectroscopy of biological fluids
Prog. Nucl. Magn. Reson. Spectrosc.
(1989) Organ toxicity of chromium in animals
Biomarkers in toxicology
Toxicology
(1998)- et al.
Rapidly progressive interstitial fibrosis in young women: association with slimming regimen including Chinese herbs
Lancet
(1993)
Spectral editing and pattern recognition methods applied to high-resolution magic-angle spinning 1H nuclear magnetic resonance spectroscopy of liver tissues
Anal. Biochem.
Application of simplified in vitro screening tests to detect genotoxicity of aristolochic acid
Food. Chem. Toxicol.
Pattern recognition classification of the site of nephrotoxicity based on metabolic data derived from proton nuclear magnetic resonance spectra of urine
Mol. Pharmacol.
Batch statistical processing of 1H NMR-derived urinary spectral data
J. Chemometr.
Using polymerase arrest to detect DNA binding specificity of aristolochic acid in the mouse H-ras gene
Carcinogenesis
Sequence-specific detection of aristolochic acid-DNA adducts in the human p53 gene by terminal transferase-dependent PCR
Carcinogenesis
Metabolic profiling of body fluids by proton NMR spectroscopy: self-poisoning episodes with paracetamol (acetaminophen)
Magn. Reson. Med.
A metabonomic investigation of hepatotoxicity using diffusion-edited 1H NMR spectroscopy of blood serum
Analyst
Effects of feeding and body weight loss on the 1H-NMR-based urine metabolic profiles of male Wistar Han rats: implications for biomarker discovery
Biomarkers
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