MOLECULAR AND FUNCTIONAL VALIDATION OF POMIFERIN AS A POTENTIAL ANTIDIABETIC AGENT: IN VITRO AND IN SILICO EVIDENCE TARGETING INSULIN SIGNALING PATHWAYS
DOI:
https://doi.org/10.22159/ijap.2026v18i4.58826Keywords:
Antidiabetic, Gene expression, In silico, In vitro, PomiferinAbstract
Objective: To investigate the molecular basis of the antidiabetic activity of Pomiferin using in vitro assays and in silico molecular docking.
Methods: Cell viability (MTT and LDH assays), glucose uptake, and enzyme activities (glycogen synthase, hexokinase, and glucokinase) were evaluated in HepG2 cells and L6 myotubes under high glucose conditions. Gene expression of insulin signaling markers (IR, IRS-1, IRS-2, AKT1, AS160, and GLUT-4) was analyzed using RT-PCR. Molecular docking was performed to determine binding energies with diabetes-associated target proteins. One-way analysis of variance (ANOVA) was used to conduct statistical analysis.
Results: High glucose reduced cell viability to 65–70% compared to control cells. Pomiferin increased cell viability to 95% at 8 µM. Glucose utilization decreased to 40–45% under high glucose and increased to 65% and 75% following treatment with 2 µM and 8 µM Pomiferin, respectively. Enzyme activities decreased under high glucose conditions [glycogen synthase: 8.5 to 4.5 nmol/min/mg protein; hexokinase: 50 to 30 mU/min/mg; glucokinase: 40 to 27 mU/min/mg] and increased following Pomiferin treatment [glycogen synthase: 6.2 nmol/min/mg protein; hexokinase: 41 mU/min/mg; glucokinase: 34 mU/min/mg]. Gene expression levels of insulin signaling markers decreased to approximately 0.4–0.6-fold under high glucose and increased to 0.8–0.9-fold following treatment. Binding energies ranged from −7.2 to −11.8 kcal/mol, with the highest affinity observed for GLUT-2.
Conclusion: Pomiferin alters the insulin signaling pathways, which supports its potential as an antidiabetic agent in the future.
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