Liquiritigenin

Osteoporosis (OP) is a prevalent systemic metabolic disease characterized by reduced bone density and compromised skeletal integrity, leading to increased fragility and fractures. This study investigated the therapeutic potential of Liquiritin (LIQ) in inhibiting osteoclastogenesis through integrated computational and experimental approaches. Molecular docking and dynamics simulations were employed to explore the interactions between LIQ and key osteoclastogenic proteins RANK and RANKL. In vitro experiments assessed LIQ's inhibitory effects on mature osteoclast (OC) formation using RNA sequencing, cellular immunofluorescence, surface plasmon resonance (SPR), cellular thermal shift assays (CETSA), and Western blot analysis. In vivo studies validated the effects of LIQ on OC formation and bone loss. The results demonstrated that LIQ (≤20 μM) exhibited no cytotoxicity to bone marrow-derived macrophages (BMMs) while potently suppressing mature OC formation. LIQ downregulated OC-specific proteins and inhibited phosphorylation in the MAPK, NF-κB, and PI3K pathways. RNA sequencing revealed that LIQ modulates mitochondrial oxidative phosphorylation and induces OC precursor apoptosis, which was confirmed by immunofluorescence and OCR/ECAR assays indicating metabolic reprogramming. Molecular docking and dynamics simulations demonstrated stable LIQ binding to RANK/RANKL, disrupting their interaction and downstream TRAF2/RIPK signaling, as verified by SPR, CETSA, and Western blot analyses. In vivo experiments confirmed that LIQ able significantly attenuated bone loss in an OVX mouse model. These findings indicate that LIQ inhibits OC formation by binding to RANK/RANKL, suppressing MAPK/NF-κB/PI3K pathway activation, and inducing metabolic reprogramming and apoptosis in OC precursors, supporting its potential as a therapeutic candidate for OP.

The development of natural anti-hyperglycemic agents with minimal side effects is a critical objective in functional food research. This study comprehensively investigated the anti-diabetic potential and bioactive substances in guava juice fermented with Lacticaseibacillus paracasei NCUH012072 (FM). Using an integrated in vitro and metabolomic approach, the anti-hyperglycemic activity was systematically assessed through a series of bioassays (enzyme inhibition, antioxidant capacity, and insulin-resistant cell models), untargeted metabolomics, and bioinformatic and affinity-based validation. The results demonstrated that FM significantly inhibited key carbohydrate-metabolizing enzymes (α-amylase, α-glucosidase) as well as the insulin signaling regulator protein tyrosine phosphatase 1B (PTP1B) (P < 0.05). Additionally, FM exhibited enhanced antioxidant capacity and improved glucose uptake in insulin-resistant HepG2 cells (P < 0.05). Untargeted metabolomics revealed that fermentation modulated 13 pivotal metabolic pathways. Integrated analysis using Spearman correlation and random forest modeling identified key anti-hyperglycemic metabolites, including flavonoids (e.g., liquiritigenin), phenolic acids (e.g., cis-caffeic acid), and redox-active compounds such as (R)-lipoic acid. Affinity ultrafiltration combined with liquid chromatography-mass spectrometry (AF-LC-MS) further characterized quercetin 3-galactoside and resveratrol as potent inhibitors of the target enzymes. This study not only systematically demonstrates the anti-hyperglycemic properties of probiotic-fermented guava juice but also proposes a comprehensive strategy for identifying bioactive compounds in complex food matrices. The findings offer a theoretical basis for developing fermented fruit-based functional foods as dietary strategies in hyperglycemia management.