Metabolite Profiling of Javanese Ginger Zingiber purpureum and Identification of Antiseizure Metabolites via a Low-Cost Open-Source Zebrafish Bioassay-Guided Isolation

Abstract

The rhizomes of Zingiber purpureum, "Bangle", were investigated for its antiseizure properties using a streamlined and cost-effective zebrafish screening strategy and a mouse epilepsy assay. Its hexane extract demonstrated strong antiseizure activity in zebrafish epilepsy assay and was, therefore, selected for bioactivity-guided fractionation. Twelve compounds (1−12) were isolated, and two bioactive phenylbutenoids, trans-(11) and cis-banglene (12), reduced up to 70% of pentylenetetrazole (PTZ)-induced seizures. These compounds showed moderate activity against PTZ-induced seizures in a mouse epilepsy assay. To understand the specificity of Z. purpureum active compounds, its chemical profile was compared to that of Z. off icinale. Their composition was assessed by differential metabolite profiling visualized by a molecular network, which revealed only vanillin derivatives and terpenoids as common metabolites and gave a comprehensive view of Z. purpureum composition. This study demonstrates the efficacy of a streamlined zebrafish epilepsy assay, which is therefore suitable for routine screening in phytochemistry laboratories.

6-Gingerol, a Major Constituent of Zingiber officinale Rhizoma, Exerts Anticonvulsant Activity in the Pentylenetetrazole-Induced Seizure Model in Larval Zebrafish

“…Instead, they noted a significant reduction of the PTZ-induced hyperlocomotion in the zebrafish larvae incubated with the hexane extract from the rhizome of Zingiber purpureum in which trans - and cis -banglene were identified as the main active compounds. As the composition of Zingiber officinale differs significantly from the herein described extract (a detailed composition was reported by the authors in the previously published manuscript [ 44 ]) it is difficult at this point to compare the anticonvulsant action of our reported Zingiber officinale extract with the previous data of Brillatz and co-workers [ 26 ]. At this point, it is important to underline that the activity of plant extracts analyzed in in vivo models may differ depending on the climatic conditions, access to water and nutrients, and so on, as shown in previous studies on different plant species [ 57 , 58 ].…”

“…In the present study, we demonstrated the anticonvulsant effect of Zingiber officinale rhizoma methanolic extract in the zebrafish larvae PTZ seizure assay. Interestingly, the study carried out in the PTZ-treated zebrafish larvae exposed to Zingiber officinale rhizome methanolic extract by Brillatz and colleagues [ 26 ] did not reveal this effect. It is worth noting that the compositional analysis of Zingiber officinale reported by these authors did not reveal the presence of 6-GIN in the extract.…”

“…Longer storage, elevated temperature, age of the plant, or location may be responsible for the transformation of gingerols to shogaols [ 59 ], which is a result of the dehydration of the molecule. These factors might have played an important role in the plant species studied by Brillatz and colleagues [ 26 ].…”

“…Previously, Zingiber officinale hydroethanolic extract was shown to exert antiseizure activity in the acute intravenous PTZ test and PTZ-kindling model in mice, but active constituents responsible for this effect have not been determined [ 24 , 25 ]. More recently, using the PTZ-induced hyperlocomotion assay in larval zebrafish, Brillatz et al [ 26 ] indicated that the hexane extract of Zingiber purpureum possesses anticonvulsant properties, with two active phenylbutenoids, i.e., trans - and cis -banglene, being responsible for this effect.…”

Bioassay-Guided Interpretation of Antimicrobial Compounds in Kumu, a TCM Preparation From Picrasma quassioides’ Stem via UHPLC-Orbitrap-Ion Trap Mass Spectrometry Combined With Fragmentation and Retention Time Calculation

“…Based on the above antimicrobial activities, it is worth performing bioassay-guided purification for identifying the antimicrobial compounds of Kumu. By now, bioassay-guided isolation of natural products have been widely used for different activities, such as anti-inflammatory (Erdemoglu et al, 2008), antioxidant (Yang et al, 2020), anti-seizure (Brillatz et al, 2020b) (Brillatz et al, 2020a), vasorelaxant (Othman et al, 2006), aldose reductase inhibitory (Logendra et al, 2006), anti-plasmodial (Baldé et al, 2021), antiviral (Parvez et al, 2020), antibacterial (He et al, 2017), and antifungal (Yan et al, 2020). But most of these studies were performed with multiple purification steps, which consumed For the bioassays, Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative), which belong to top dangerous superbugs worldwide (World Health Organization, 2017), were selected for the antimicrobial activities of Kumu.…”