Network pharmacology and metabolomics reveal mathurameha, a Thai traditional Anti-Diabetic formula, enhances glucose metabolism through PI3K-AKT/AMPK/GLUT4 pathway modulation.

Traditional herbal formulations offer promising avenues for diabetes management by targeting multiple molecular pathways. Mathurameha (MT), a polyherbal preparation, has been historically used for its antidiabetic potential. However, its molecular mechanisms remain largely unexplored. FrE exhibited potent α-glucosidase inhibition (IC₠₀ 0.3 µg/mL) and significantly enhanced glucose uptake in L6 myotubes (3.67 ± 0.23-fold) and 3T3-L1 adipocytes (IC₠₀ 6.78 µg/mL). It also stimulated insulin secretion (1.42-fold), comparable to metformin (1.46-fold), and protected INS-1 pancreatic β-cells from H₂O₂-induced apoptosis (30.65 ± 3.54%) through partial caspase-3 inhibition. LC-MS-QTOF analysis identified 73 metabolites, including ellagic acid, kushenol A, gallic acid, arctiin, neoandrographolide, astilbin, paenol, muricatacin, coumarrayin, and zingerone. Network pharmacology and pathway enrichment analyses revealed key targets (GSK3β, GLUT4, PPARG, INSR, AKT2, CASP3, and MMP9) and highlighted the involvement of PI3K-AKT, AMPK, and GLUT4 signaling pathways. Gene expression analysis confirmed the upregulation of GLUT4, AMPK, IRS, PI3K, and AKT genes in L6 myotubes treated with FrE. These findings suggest that MT exerts antidiabetic effects via the PI3K-AKT/AMPK/GLUT4 signaling axis, promoting glucose uptake, insulin secretion, and β-cell protection. Future studies will focus on in vivo validation, standardization of bioactive fractions, and omics-based approaches to establish a well-defined, effective formulation for diabetes management.