A Chemically Characterized Polyherbal Extract Selectively Attenuates Chronic Unpredictable Stress-Induced Redox-Sensitive, Inflammatory, and Metabolic Alterations in Female Rats.

Chronic stress contributes to metabolic and mental disorders, with accumulating evidence suggesting differences in stress responses between females and males. Although Centaurium erythraea, Valeriana officinalis, Melissa officinalis, and Humulus lupulus possess well-documented antioxidant, anti-inflammatory, and neuroactive properties, the biological activity of their combined extract remains largely unexplored. Here, a chemically characterized polyherbal extract was evaluated for in vitro biosafety and its effects on chronic unpredictable stress-induced behavioral, metabolic, and molecular alterations in male and female rats. LC-HRMS/MS identified 121 metabolites, including phenolic acids, iridoid glycosides, flavonoids, and xanthone derivatives, while the extract showed a favorable biosafety profile in HaCaT keratinocytes. Chronic stress-induced alterations were predominantly observed in females, including reduced body mass, serum glucose and insulin levels, enhanced hippocampal glucocorticoid signaling, increased NRF2 and NFκB protein levels, altered IRS1 signaling, and anxiety-like behavior. Treatment with the extract normalized glucose tolerance, restored hippocampal IRS1, phosphorylated IRS1 (Ser307), NRF2, and NFκB protein levels in females. These findings indicate that, under the experimental conditions employed, chronic stress-associated alterations and the effects of the polyherbal extract were predominantly observed in female rats, and suggest that the extract may mitigate stress-induced metabolic disturbances through modulation of redox-sensitive, inflammatory, and hippocampal insulin signaling pathways.