Jiamusi

Isofraxidin, a hydroxylated coumarin occurring in traditional medicinal plants such as Eleutherococcus senticosus, has been widely investigated for anti-inflammatory, antioxidant and neuroprotective actions in adult models, yet its developmental safety is unclear. Wild-type AB zebrafish embryos were exposed to isofraxidin from 6 to 96 hpf. Survival was monitored to calculate the 96 hpf LC₅₀, and developmental endpoints (malformations, body length, heart rate and yolk sac area) were assessed at 96 hpf. Neurobehavior was quantified by spontaneous tail coiling and locomotor tracking. Neurodevelopment was evaluated using brain ventricle morphology, neuronal fluorescence in Tg(elavl3:EGFP) larvae, acetylcholinesterase activity, and qRT-PCR of neurodevelopment-related genes. Network toxicology was applied to predict intersecting targets and enriched pathways, followed by validation of oxidative stress (ROS, SOD, CAT, GSH and MDA) and apoptosis (acridine orange staining and apoptosis-related transcripts). Isofraxidin caused dose dependent developmental toxicity, reducing survival and increasing malformations at higher concentrations. Sublethal exposure produced robust neurobehavioral deficits, with decreased tail coiling and reduced locomotor activity. Concordantly, isofraxidin impaired brain ventricle development, reduced neuronal fluorescence, decreased AChE activity, and dysregulated neurodevelopmental gene expression. Mechanistically, network toxicology highlighted oxidative stress and apoptosis-associated pathways; experimentally, isofraxidin increased ROS accumulation, suppressed antioxidant defenses (lower SOD, CAT and GSH), elevated lipid peroxidation (higher MDA), and enhanced apoptotic signals in the brain region. These findings demonstrate developmental neurotoxicity of isofraxidin in zebrafish and support oxidative stress and apoptosis as a key mechanism, emphasizing the need to evaluate early-life safety ethnopharmacological bioactives.

Physalis alkekengi L. (P. alkekengi) is widely used in traditional medicine and functional foods for its various medicinal properties. However, its toxicological profile has not been comprehensively assessed.

This study aimed to characterize the developmental toxicity and cardiotoxicity of P. alkekengi using a Danio rerio (D. rerio) embryo model and to identify key toxic constituents to support safety optimization.

D. rerio embryos were exposed to the fruit and calyx extracts of P. alkekengi as well as membrane-fractionated preparations to assess toxic phenotypes. Chemical profiling was performed using ultra-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-Exactive Orbitrap-MS), and mechanistic insights were explored using apoptosis-related assays and network toxicology analysis.

The fruit extract of P. alkekengi exhibited stronger developmental toxicity and cardiotoxicity effects compared to the calyx extract. The 5-10 kDa non-polysaccharide fraction of the fruit was identified as the primary toxic source, with Glycidyl stearate (GS) being the key cardiotoxic component in this fraction. Removal of the most toxic 5-10 kDa fraction during the extraction process significantly reduced the toxicity of the fruit extract.

The toxicity of P. alkekengi fruit extract is concentrated in the non-polysaccharide 5-10 kDa fraction, with GS being the main contributor to the toxicity in this fraction. Membrane-based separation technology to remove this fraction can enhance the safety of the extract. This study provides a framework for the safety evaluation and optimization of traditional medicinal plants.

Employing a comprehensive strategy that integrated network pharmacology, molecular docking, and in vitro experimental techniques, this study examined the effect of neohesperidin on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) upon lipopolysaccharide (LPS)-induced inflammatory conditions.

Core genes were screened using molecular docking and network pharmacology. hPDLSCs were isolated and cultured, and their surface markers were identified by flow cytometry. The effects of neohesperidin on hPDLSC proliferation and osteogenic differentiation upon LPS stimulation were investigated using ELISA, CCK-8, alkaline phosphatase, alizarin red staining, qRT-PCR, and Western blot analyses.

hPDLSCs highly expressed CD90 but did not express CD34 and CD45, which is consistent with the characteristics of stem cells. Network pharmacology and molecular docking analyses revealed that inflammatory and osteogenesis-related signaling pathways were significantly enriched. Treatment with 50 μM neohesperidin promoted proliferation, alkaline phosphatase activity, and mineralized nodule formation while suppressing the inflammatory response in LPS-stimulated hPDLSCs (P < 0.05). It also upregulated the expression of osteogenic markers, such as Runx family transcription factor 2 (RUNX2), osteocalcin (OCN), and osterix (OSX) (P < 0.05). Furthermore, neohesperidin activated the PI3K/AKT signaling pathway and reversed the inhibitory effect of LY294002 (P < 0.05).

Through PI3K/AKT activation, neohesperidin stimulated the osteogenic transformation of hPDLSCs under inflammatory conditions, suggesting that it may be a pharmacologically active compound with the potential for use as an adjuvant in periodontitis treatment.