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Objectives: Nowadays, the global population is moving towards herbal drugs, which contain bioactive compounds, to cure diseases. Rauwolfia serpentina is a medicinally important herb that is mainly effective in the treatment of hypertension and psychotic disorders. The present study was designed to investigate the effects of Rauwolfia serpentina on acute stress. The herb extract was orally administered before immobilization for 2 h only, to monitor any change in behavioral activities. We also evaluated the role of Rauwolfia serpentina in oxidative stress, including its effect on antioxidant enzymes’ activities, such as catalase and superoxide dismutase, and also on plasma glucose, corticosterone and leptin levels. Methods: Animals were divided into four groups, which were (i) saline unstressed, (ii) Rauwolfia serpentina unstressed, (iii) saline stressed and (iv) Rauwolfia serpentina stressed, which were injected accordingly with saline (1 mL/kg) or Rauwolfia serpentina (30 mg/kg). Animals of the stressed group received immobilization for 2 h. Behavioral analysis was performed after the termination of the 2 h immobilization period. Animals were then decapitated and plasma samples were collected for CAT, SOD, corticosterone, leptin and glucose estimation. Results: Results showed that Rauwolfia serpentina is an effective anxiolytic agent as it attenuates stress-induced behavioral deficits and improves locomotor activity. On the other hand, it provides positive outcomes regarding the antioxidant enzymes levels of stressed animals. Conclusion: Rauwolfia Serpentina was found to prevent the stress-induced increase in corticosterone, and an increase in the levels of endogenous leptin attenuates the stress-induced activity of the HPA axis. It is also concluded that 30 mg/kg of Rauwolfia serpentina is not sufficient to produce hypoglycemic effects. However, more studies are recommended to explain the particular action by which Rauwolfia serpentina produces its effects. Full article