Mussaenda erythrophylla Exhibits Anxiolytic-Like Activity in Healthy Rats in India: A Neurobehavioral, Neurochemical, and Network Pharmacology Study

Background: Anxiety disorders affect approximately 4.4% of the global population, highlighting the need for alternative therapeutic strategies. Medicinal plants containing structurally diverse bioactive constituents have attracted interest as potential sources of multitarget anxiolytic agents. Methods: The anxiolytic-like activity of the ethanolic leaf extract of Mussaenda erythrophylla Schumach. & Thonn. (Rubiaceae) was investigated using phytochemical screening, liquid chromatography–mass spectrometry, network pharmacology, molecular docking, and behavioral, neurochemical, and histopathological assessments. Healthy Wistar rats (n = 6/group) received vehicle (0.5% sodium carboxymethyl cellulose), diazepam (1 mg/kg), or extract (100, 200, or 400 mg/kg) orally for 14 days. Behavioral responses were evaluated using five established anxiety-related paradigms without experimentally inducing anxiety. Results: Phytochemical screening detected flavonoids, phenolics, terpenoids, alkaloids, and glycosides. Liquid chromatography–mass spectrometry tentatively identified six constituents, including quercetin, apigenin-7-glucoside, oleanolic acid, and campesterol. Network pharmacology identified AKT1, TNF-α, IL-6, STAT3, and GAPDH as predicted hub targets, while molecular docking suggested potential ligand–target interactions. The extract produced dose-dependent anxiolytic-like behavioral responses, with the highest dose approaching diazepam-associated effects in several measures. Neurochemical assays indicated increased GABA, serotonin, and dopamine concentrations and reduced glutamate concentrations. Histopathological examination suggested dose-associated differences in neuronal morphology. Discussion: The behavioral and neurochemical findings are consistent with potential involvement of multiple neurotransmitter systems, whereas the computational analyses provide hypotheses concerning inflammatory and intracellular signaling pathways that require experimental validation. Conclusion: Mussaenda erythrophylla ethanolic leaf extract demonstrated anxiolytic-like activity in healthy rats. Further investigations using validated anxiety-induction models, analytically specific neurochemical assays, and mechanistic experiments are necessary to establish its therapeutic potential.