Screening of potential antiepileptic active ingredients in Rhizoma Gastrodiae based on zebrafish model and metabolomics technology
Identifying potential antiepileptic active ingredients in Rhizoma Gastrodiae is of immense significance for the prevention and treatment of epilepsy.In this study,we used a larval zebrafish epilepsy model to evaluate the antiepileptic activity of Rhizoma Gastrodiae from two different regions.In addition,we employed metabolomics technology based on liquid chromatography tandem mass spectrometry/QE plus to detect the ingredients in Rhizoma Gastrodiae and performed pattern analysis to identify key differential metabolites.The metabolites were further identified using primary and secondary mass spectrometry data and literature references.This step was followed by the confirmation of their antiepileptic activity using the zebrafish epilepsy model.The results showed that nine extracts of Rhizoma Gastrodiae significantly reduced the number of whirls in zebrafish,and significant differences in antiepileptic activity were observed between the Rhizoma Gastrodiae samples from the two regions(P<0.05).Metabolomics and pattern analysis identified six important differential metabolites(Parishin E,Gastrodin,Parishin C,Parishin D,N6-p-hydroxybenzyl adenosine,and 4,4′-dihydroxydibenzyl ether).Activity verification results showed that Parishin E,Gastrodin,and N6-p-hydroxybenzyl adenosine significantly inhibited zebrafish epilepsy-like behavior.This study utilized the zebrafish model and metabolomics to identify and determine several active antiepileptic ingredients in Rhizoma Gastrodiae.Of these,Parishin E and N6-p-hydroxybenzyl adenosine were reported for the first time to exhibit antiepileptic activity,thereby serving as a valuable reference for further research on the antiepileptic effects of Rhizoma Gastrodiae.
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