Integrated metabolomics and network pharmacology reveal multi-target phytotherapeutic activities of Frankenia hirsuta against inflammation and diabetic neuropathy with cytotoxic potential

OBJECTIVES: Frankenia hirsuta (FH) is an underexplored halophytic plant used for managing inflammatory and metabolic disorders. However, its phytochemical composition and pharmacological mechanisms remain largely uncharacterized. Thus, this study aimed to comprehensively elucidate the phytochemical profile, molecular mechanisms, and pharmacological efficacy of FH using an integrated approach combining liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolite characterization, in vitro bioassays, in-vivo validation, and computational analyses. METHODS: Metabolite profiling was performed using LC-MS/MS, followed by molecular docking and network pharmacology analyses to predict potential targets involved in neuropathic, inflammatory, and metabolic pathways. In vitro cytotoxicity and anticoagulant activities of the aerial and root ethanolic extracts were assessed using MTT and activated partial thromboplastin time (aPTT) assays, respectively. The in vivo antidiabetic and neuroprotective effects were evaluated in streptozotocin-induced diabetic mice, using behavioral assays (hot plate and von Frey filaments), biochemical markers of oxidative stress and inflammation, and histopathological and immunohistochemical (IHC) examinations. KEY FINDINGS: LC-MS/MS analysis revealed a diverse bioactive constituents, including phenolic acids, flavonoid glycosides, and feruloyl derivatives. Network pharmacology and docking studies highlighted strong binding affinities toward key enzymes such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), cyclooxygenase-2, and AKT1, indicating multimodal anti-inflammatory and neuroprotective potential. In vitro, FH extracts exhibited significant cytoprotective effects and prolonged aPTT in a dose-dependent manner, confirming their safety and mild anticoagulant properties. In vivo, FH root extract (75 mg/kg) and its isolated feruloyl glycoside (FG) (5-10 mg/kg) significantly restored nociceptive thresholds, improved glycemic control, normalized oxidative and inflammatory biomarkers, and preserved pancreatic and sciatic nerve histoarchitecture. IHC analyses further demonstrated downregulation of TNF-α and IL-6 and upregulation of IL-10 in treated groups, confirming their anti-inflammatory and regenerative effects. CONCLUSIONS: This study provides the first comprehensive evidence that FH exerts potent neuroprotective and antidiabetic effects mediated through antioxidant, anti-inflammatory, and cytokine-modulating pathways. The integrated LC-MS/MS, computational, and experimental validation framework highlights FH and its FG as promising candidates for developing safe, plant-derived therapeutics targeting diabetic neuropathy and related metabolic dysfunctions.