Metabolomics Study of Aristolochic Acid Nephrotoxicity in Rodents
| Autor: | Dong-Ming Tsai, 蔡東銘 |
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| Rok vydání: | 2009 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 97 Aristolochic acid (AA) is a potent nephrotoxic agent that can be found in several herbs of the genus Aristolochia. It was once commonly used in traditional Chinese medicine remedy. In this study, we applied our metabolomics platform of the 1H NMR spectroscopy on urine with principal component analysis (PCA) to detect and further dissect the nephrotoxicity in rodent of four AA containing materials, AA standard, Madouling, an AA containing herb Aristolochia contorta and Bu-Fei-A-Jiau-Tang (BFAJT), a herb compound containing Aristolochia contorta. The aim was to use the easy available urine samples to establish a model for early diagnosis of nephrotoxicity. The experiment was divided into four parts. The first was rat AA experiment. The second was mouse AA experiment. The third was mouse Madouling experiment. The fourth was mouse BFAJT experiment. Urine samples were collected daily and freeze-dried for storage. All animals were euthanized after experiment and their kidneys and livers procured for pathological studies. Results: In the rat AA experiment, pathology showed grade (gr.) 1-3 acute renal tubulointerstitial necrotic change (ATIN) after 5 doses in the high dose group. In the mouse AA experiment, 10 doses were given. Both high and low dose group showed gr. 3-4 ATIN. In the mouse Madouling experiment, 21 doses were given. It was gr. 1-2 for low and moderate groups, and gr. 3-4 ATIN for the high dose group. In the BFAJT experiment, 20 doses were given. It was gr. 2 ATIN in the low dose group and gr. 3-4 in the high dose group. PCA scoring plots for the urine NMR spectral chemical shifts variables showed early cluster at 2 day and later for each group in the rat AA experiment. In the mouse AA experiment, the high dose group was clustered from the other 2 groups at day 8. It was at day 10 that the 2 dose groups were clustered from the control group. In the mouse Madouling experiment, the high dose group clustered from all other 3 groups. It was at day 10 that the high dose and the moderate dose groups clustered together from the other 2 groups. In the BFAJT experiment, PCA scoring plots failed to classify across control and dose groups even at day 16. The endogenous metabolites assigned from NMR spectroscopy and their integral concentration among dose and control groups were tested by t-test. In the rat AA experiment glycine, succinate, 2-oxoglutarate and trimethylamine-N-oxide (TMAO) were decreased in the dose groups in earlier days of the experiment. At later days, sugar, allantoin, creatine, dimethylglycine, 2-oxoglutarate and TMAO were increased in the dose groups (p < 0.05). In the mouse AA experiment no significant metabolite difference was noted, only alanine, lactate and formate had more than two fold change at day 10. In conclusion, AA standard, Madouling and BFAJT were all nephrotoxic. PCA scoring plots for the urine NMR spectroscopy collected from living animals showed the ability to classify the dose and control groups days before confirming the renal pathology. But toxicity classifying failed in the mouse BFAJT experiment by PCA scoring plot. Further metabolomics study is expected to resolve this issue. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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