Τίτλος:
Antiseizure potential of the ancient Greek medicinal plant Helleborus odorus subsp. cyclophyllus and identification of its main active principles
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
Ethnopharmacological relevance: Ethnopharmacological data and ancient texts support the use of black hellebore (Helleborus odorus subsp. cyclophyllus, Ranunculaceae) for the management and treatment of epilepsy in ancient Greece. Aim of the study: A pharmacological investigation of the root methanolic extract (RME) was conducted using the zebrafish epilepsy model to isolate and identify the compounds responsible for a potential antiseizure activity and to provide evidence of its historical use. In addition, a comprehensive metabolite profiling of this studied species was proposed. Materials and methods: The roots were extracted by solvents of increasing polarity and root decoction (RDE) was also prepared. The extracts were evaluated for antiseizure activity using a larval zebrafish epilepsy model with pentylenetetrazole (PTZ)-induced seizures. The RME exhibited the highest antiseizure activity and was therefore selected for bioactivity-guided fractionation. Isolated compounds were fully characterized by NMR and high-resolution tandem mass spectrometry (HRMS/MS). The UHPLC-HRMS/MS analyses of the RME and RDE were used for dereplication and metabolite profiling. Results: The RME showed 80% inhibition of PTZ-induced locomotor activity (300 μg/ml). This extract was fractionated and resulted in the isolation of a new glucopyranosyl-deoxyribonolactone (1) and a new furostanol saponin derivative (2), as well as of 20-hydroxyecdysone (3), hellebrin (4), a spirostanol glycoside derivative (5) and deglucohellebrin (6). The antiseizure activity of RME was found to be mainly due to the new furostanol saponin (2) and hellebrin (4), which reduced 45% and 60% of PTZ-induced seizures (135 μM, respectively). Besides, the aglycone of hellebrin, hellebrigenin (S34), was also active (45% at 7 μM). To further characterize the chemical composition of both RME and RDE, 30 compounds (A7-33, A35–37) were annotated based on UHPLC-HRMS/MS metabolite profiling. This revealed the presence of additional bufadienolides, furostanols, and evidenced alkaloids. Conclusions: This study is the first to identify the molecular basis of the ethnopharmacological use of black hellebore for the treatment of epilepsy. This was achieved using a microscale zebrafish epilepsy model to rapidly quantify in vivo antiseizure activity. The UHPLC-HRMS/MS profiling revealed the chemical diversity of the extracts and the presence of numerous bufadienolides, furostanols and ecdysteroids, also present in the decoction. © 2020 The Authors
Συγγραφείς:
Brillatz, T.
Jacmin, M.
Vougogiannopoulou, K.
Petrakis, E.A.
Kalpoutzakis, E.
Houriet, J.
Pellissier, L.
Rutz, A.
Marcourt, L.
Queiroz, E.F.
Crawford, A.D.
Skaltsounis, A.-L.
Wolfender, J.-L.
Περιοδικό:
Journal of Ethnopharmacology
Εκδότης:
Elsevier Ireland Ltd
Λέξεις-κλειδιά:
21 hydroxymethyl 24 [(o beta dextro glucopyranosyl(1-4)beta dextro fucopyranosyl)oxy] 3beta,23 dihydroxyspirosta 5,25 (27)diene 1beta yl o (alpha levo rhamnopy ranosyl)(1-2) o [beta dextro xylopyranosyl(1-3)]alpha levo arabinopyranoside; 4 hydroxy 5 [(beta dextro glucopyranosyloxy)methyl]dihydrofuran 2(3h) one; aglycone; alkaloid; anticonvulsive agent; bufadienolide; deglucohellebrin; ecdysterone; furostanol; hellebrigenin; hellebrin; helleodoraside; plant extract; plant glycoside; saponin derivative; topiramate; unclassified drug; anticonvulsive agent; methanol; pentetrazole; phytochemical; plant extract; solvent, animal experiment; animal model; anticonvulsant activity; Article; biological activity; chemical composition; controlled study; drug identification; drug isolation; drug screening; drug synthesis; embryo; epilepsy; fish model; fractionation; Helleborus; Helleborus odorus subsp. cyclophyllus; locomotion; medicinal plant; metabolic fingerprinting; nonhuman; nuclear magnetic resonance spectroscopy; pentylenetetrazole-induced seizure; plant root; solvent extraction; tandem mass spectrometry; ultra performance liquid chromatography; zebra fish; animal; chemistry; comparative study; disease model; dose response; drug effect; high performance liquid chromatography; isolation and purification; metabolism; metabolome; metabolomics; pathophysiology; seizure, Animals; Anticonvulsants; Chromatography, High Pressure Liquid; Disease Models, Animal; Dose-Response Relationship, Drug; Helleborus; Locomotion; Metabolome; Metabolomics; Methanol; Pentylenetetrazole; Phytochemicals; Plant Extracts; Plant Roots; Seizures; Solvents; Tandem Mass Spectrometry; Zebrafish
DOI:
10.1016/j.jep.2020.112954
