Application of metabonomic strategy to discover an unreported active ingredient in LiuWeiDiHuang pills suppressing beta-glucuronidase.
Identification of the bioactive ingredient from traditional Chinese medicine (TCM) remains a challenging task by traditional approach that focuses on chemical isolation coupled with biological activity screening. Here, we present a metabonomics-based approach for bioactive ingredient discovery in LiuWeiDiHuang pills (LWPs). First, a non-targeted high-performance liquid chromatography ultraviolet (HPLC-UV) profiling of rat urine was used to discriminate urinary profiling intervened by LWPs. Orthogonal partial least-squares discriminant analysis (OPLS-DA) revealed that eight chromatographic peaks made a significant contribution to the classification of the LWPs group and the control group. Five of these chromatographic peaks were successfully isolated and identified as hippurate, genistein (GT), daidzein (DZ), and glucuronide conjugate of GT and that of DZ by mass spectroscopy (MS). Subsequently, we found that LWPs significantly decreased the activity of intestinal β-glucuronidase by 18 % and exerted a dose-dependent inhibitory effect on rat liver lysosomal fraction, suggesting that LWPs were a β-glucuronidase inhibitor. In the end, by inhibiting β-glucuronidase-guided isolation, D-glucaro-1,4-lactone, a previously unreported ingredient of LWPs, was identified by MS, MS/MS, and nuclear magnetic resonance spectroscopy. Our findings indicated that metabonomics might increase research productivity toward the drug targets and/or bioactive compounds from TCM.
School of Pharmacy, Nanchang University, Nanchang, 330006, China, xiebaogang@ncu.edu.cn.
Baogang
B
Xie
Zhirong
Z
Zhang
Tao
T
Gong
Ningning
N
Zhang
Huiyun
H
Wang
Huiqing
H
Zou
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D016002:Discriminant Analysis; D004365:Drugs, Chinese Herbal / Q000032:analysis / Q000737:chemistry* / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000032:analysis / Q000494:pharmacology; D005937:Glucaric Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology; D005966:Glucuronidase / Q000037:antagonists & inhibitors*; D016018:Least-Squares Analysis; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D055432:Metabolomics / Q000379:methods*; D017207:Rats, Sprague-Dawley; D016482:Urinalysis / Q000379:methods
D015415:Biomarkers; D004365:Drugs, Chinese Herbal; D004791:Enzyme Inhibitors; C042579:Liuwei Dihuang Decoction; C009194:saccharolactone; D005966:Glucuronidase; D005937:Glucaric Acid