Hepatotoxic constituents and toxicological mechanism of Xanthium strumarium L. fruits.
BACKGROUND
In the recent years, the international community has attached increasing importance to possible toxicity associated with Traditional Chinese Medicine (TCM). And hepatotoxicity is one of the major concerns, a fundamental pathological process induced by toxicant. This paper is in an attempt to identify the hepatotoxic components in Xanthium strumarium L. fruits (XSF) and interpret the toxicological mechanism induced by XSF.


METHODS
XSF extract was prepared and seven characteristic components were isolated and identiﬁed in XSF water extracts. We evaluated their hepatotoxicity effect on cell proliferation and lactate dehydrogenase (LDH) activity in L-02 and BRL liver cell line. An integrated metabonomics study using high-resolution (1)H nuclear magnetic resonance ((1)H NMR) spectroscopy combined with multivariate statistical analysis was undertake to elucidate the hepatotoxicity mechanism induced in rats by XSF. The urine and serum metabolites were measured after treatment of rats with XSF (7.5, 15.0 and 30.0 g/kg/day) for 5 days.


RESULTS
The results showed that atractyloside, carboxyatractyloside, 4'-desulphate-atractyloside and XSF induced significant cytotoxic effects in both L-02 and BRL liver cell lines, indicating that atractyloside, carboxyatractyloside, and 4'-desulphate-atractyloside were the toxic components of XSF. When rats were treated with XSF at 30.0 g/kg the hepatotoxicity was reflected in the changes observed in serum biochemical profiles and by the histopathological examination of the liver. The levels of VLDL/LDL, 3-HB, lactate, acetate, acetone and glutamate in serum were increased in this group, while d-glucose, choline and valine were decreased. The elevation in the levels of succinate, citrate, 2-oxo-glutamate, glycine, 3-HB, acetate, lactate, hippurate, dimethylglycine, methylamine, dimethylamine, phenylalanine and tryptophan was observed in urine, in contrast a reduction in the intensities of taurine, d-glucose, N-acetyl-glucoprotein and trimethylamine-N-oxide (TMAO) was observed.


CONCLUSIONS
The results demonstrate that the major hepatotoxicity constituents are atractyloside, carboxyatractyloside and 4'-desulphate-atractyloside, and the hepatotoxicity of XSF involves mitochondrial inability, fatty acid metabolism, and some amino acids metabolism. This integrated (1)H NMR -based metabolic profiling approach has been able to capture and probe the metabolic alterations associated with the onset and progression of hepatotoxicity induced by XSF, and permits a comprehensive understanding of systemic toxicity for phytochemicals and other types of xenobiotic agents.
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China; Health Laboratory, Shanghai Municipal Center for Disease Control and Prevention, 1380 North Zhongshan Road, Shanghai 200336, People's Republic of China.
Li-Ming
LM
Xue
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China.
Qiao-Yan
QY
Zhang
Center for Disease Control and Prevention, Jinan Military Region, PLA, 36 East Wenhua Road, Jinan, Shandong 250012, People's Republic of China.
Ping
P
Han
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China.
Yi-Ping
YP
Jiang
The Second Affiliated Hospital of Shandong Traditional Chinese Medicine University, 1 Jingba Road, Ji'nan 250001, People's Republic of China.
Rong-Di
RD
Yan
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China.
Yang
Y
Wang
Faculty of Science, School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK.
Khalid
K
Rahman
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China.
Min
M
Jia
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China. Electronic address: than927@163.com.
Ting
T
Han
Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, People's Republic of China. Electronic address: qinsmmu@126.com.
Lu-Ping
LP
Qin
D000818:Animals; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005638:Fruit; D006801:Humans; D008099:Liver / Q000187:drug effects* / Q000473:pathology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008516:Medicine, Chinese Traditional / Q000009:adverse effects; D055432:Metabolomics; D015999:Multivariate Analysis; D010936:Plant Extracts / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D031210:Xanthium / Q000737:chemistry*
D010936:Plant Extracts
(1)H NMR; Hepatotoxicity; Metabonomics; Xanthium strumarium L.