To date, exploration of Gracilaria and Gelidiella’s bioactive compounds has been conducted using conventional methods that require a long time, high costs, and significant effort. Currently, metabolomic profiling and molecular networking have emerged as methods of exploring bioactive compounds. This study aimed to perform bioactive compound profiling through a metabolomic LC-HRMS-based and molecular networking approach in Gelidiella acerosa, Gracilaria arcuata, and Gracilaria verrucosa. All chromatograms and MS/MS spectra obtained for three crude extracts were digitally converted into an mzXML file using MSConvert, submitted to the Global Natural Product Social (GNPS), and visualized in Cytoscape 3.9.1. In total, nine dereplicated compounds were identified: 11-Deoxyprostaglandin (m/z 324.214), Diacylglyceryl trimethylhomoserines (DGTS) (m/z 684.575), Glycochenodeoxy acid (m/z 448.369), Lysophosphatidylcholine (m/z 522.350), Diacylglyceryl trimethylhomoserines (DGTS) (m/z 656.557), Pheophorbide A (m/z 593.266), Pyropheophorbide A (m/z 593.266), (2R,3R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-2,3-dihydro-4H-chromen-4-one (m/z 303.15), and Polyporic acid (m/z 293.156). These compounds are typically classified as fatty acids, lipids, terpenoids, alkaloids, shikimates, and phenylpropanoids. The molecular networking and metabolite clustering showed an interesting pattern where some compounds were produced only by one species, some by two species, and some by all three. These compounds may have pharmaceutical potential based on their chemical properties and reported activities.

Flammulina velutipes (enoki mushroom) is a functional edible mushroom rich in antioxidants, polysaccharides, mycosterols, fiber, and minerals. Accumulating evidence highlights its therapeutic potential across diverse pathological contexts, including boosting cognitive function. However, its role in neuromodulation has not been systematically explored. This study examined the effects of methanolic and ethanolic extracts of F. velutipes on primary hippocampal neurons. Neurons were treated with different extract concentrations, followed by assessments of cell viability, cytoarchitecture, neuritogenesis, maturation, and neuroprotection under oxidative stress. The extracts were further characterized by GC-MS to identify bioactive metabolites, and molecular docking combined with MM-GBSA binding energy analysis was employed to predict potential modulators. Our results demonstrated that the methanolic extract significantly enhanced neurite outgrowth, improved neuronal cytoarchitecture, and promoted survival under oxidative stress, whereas the ethanolic extract produced moderate effects. Mechanistic studies indicated that these neuroprotective and neurodevelopmental benefits were mediated through activation of the NTRK receptors, as validated by both in vitro assays and molecular docking studies. Collectively, these findings suggest that F. velutipes extracts, particularly methanolic fractions, may serve as promising neuromodulatory agents for promoting neuronal development and protecting neurons from oxidative stress.

Bacopa monnieri (L.) Wettst. (Family: Scrophulariaceae) is a well-known edible plant used in ethnic and Ayurveda medicine for centuries to improve memory deficit, enhance cognitive function, and treat nervous system disorders. Despite accumulating in vivo evidence for its cognitive benefits, the detailed mechanisms by which its bioactive compounds act on primary neurons remain elusive. In the present study, we dissect the mechanism by which Bacopa monnieri promotes neuronal development by treating primary hippocampal neuronal cultures with its ethanolic extract (BMEE) and integrating insights from in silico network pharmacology. We identified that BMEE at different concentrations promotes neuritogenesis and has a remarkable impact on early neuronal maturation, and axonal and dendritic outgrowth. Also, BMEE regulated synaptic plasticity by increasing the expression of NMDA receptors. Metabolites of BMEE were identified by gas chromatography–mass spectrometry (GC-MS) analysis, from which a network pharmacology model was constructed, in which BMEE metabolites were projected to regulate the neurotrophin signaling pathway. Indeed, the BMEE-mediated neuritogenic effect was abolished by the presence of a TrkA receptor-specific inhibitor, suggesting that the neuritogenic effect of BMEE is TrkA-dependent. Also, molecular docking following MD simulations supported the idea that BMEE metabolites, particularly δ-Tocopherol and O-methyl-, bind with high affinity to the TrkA receptor (NGF-binding domain). This study collectively illuminates the TrkA-mediated pathway through which Bacopa monnieri promotes neuronal development and suggests that bioactive metabolites from BMEE might hold potential as a source for designing therapeutic agents for various cognitive disorders.

Flammulina velutipes is a well-known edible mushroom which is consumed for its nutri-tional value. F. velutipes is also known as enoki mushroom, a good source of antioxidants, fiber, polysaccharides, mycosterol and minerals that make it a nutritious functional food. In this present study we utilized methanol and ethanol extract of F. velutipes mushroom on primary neuronal cultured cell and made a comparative study on neuronal develop-ment, cytoarchitecture, neuronal viability and maturation. We also characterized the methanolic and ethanolic extracts of mushroom with GC-MS profiling and detected the active metabolites present in the extracts. The comparative study showed methanolic ex-tract of the mushroom has more promising impact on neuronal development and neuri-togenesis. Furthermore, we evaluated the mechanistic insight and found that the impact of the mushroom is dependent on NTRK receptor. Additionally, we validated the activa-tion of NTRK receptor by in-silico molecular docking study.