Abstract
Zebrafish, a common model organism for studies of vertebrate development and gene function, has been used in pharmaceutical research as a new and powerful tool in recent years. In the present study, we applied zebrafish for the first time in a metabolic study of notoginsenoside (R1), ginsenoside (Rg1) and ginsenoside (Rb1), which are saponins isolated from Panax notoginseng. Metabolites of these three saponin compounds in zebrafish after exposure for 24 h were identified by high performance liquid chromatography - electrospray mass spectrometry (HPLC-ESI-MS) with a Zorbax C-18 column for separation using a binary gradient elution of 0.05% formic acid acetonitrile - 0.05% formic acid water. The quasi-molecular ions of compounds were detected in negative mode. Step-wise deglycosylation metabolites and hydroxylation metabolites of the three saponins were found, which were coincide with regular methods for metabolic analysis. Our study demonstrated that the zebrafish model can successfully imitate the current metabolic model with advantages of lower cost, far less amount of compound needed, easy set up and high performance. Our data suggests that the zebrafish metabolic model has the potential for developing a novel method for quickly predicting the metabolism of Chinese herb components, including those of trace compounds.
Highlights
In recent years, metabolic studies of Traditional Chinese Medicine (TCMs) have been applied effectively in the major fields of TCM research, including material basis, mechanism of action and quality control
Our present study found deglycosylation and hydroxylation of R1, Rg1 and Rb1 by zebrafish for the first time, which is highly consistent with the results of existing methods: all stepwise deglycosylation metabolites were found, except protopanaxatriol and protopanaxadiol, the reasons maybe that stable monoglucosylated ginsenoside is difficult to degrade further [30]; hydroxylation of R1 and Rb1 were totally consistent with the rat metabolism data [15,28], and hydroxylation of Rg1 found for the first time provides supporting evidence for the presumption of reference [29]
Adult zebrafish were divided into four experimental groups of five fish each after fasting for 12 h one blank control group was exposed to 1% dimethyl sulfoxide (DMSO) purified water, three groups were exposed to 30 mL solution of R1 (9.63 μg/mL), Rg1 (16.75 μg/mL) and Rb1 (21.24 μg/mL)
Summary
Metabolic studies of Traditional Chinese Medicine (TCMs) have been applied effectively in the major fields of TCM research, including material basis, mechanism of action and quality control. Over the past ten years, zebrafish has been used as a popular model organism in diverse fields of research such as developmental and evolutionary biology, toxicology and pharmacology They are considered a new powerful tool, especially in pharmaceutical research [1,2,3,4,5,6,7], because they are genetically similar to humans and have the same complex organs found in mammals. Based on results from zebrafish metabolic study model and existing in vivo and in vitro study methods, we report evidence for a feasible and economic model, and our research should contribute to developing a novel, simple, low cost, high-performance metabolic study model for trace components of TCMs
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