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<p>Abstract</p> <p>Background</p> <p>Compound <it>Valeriana jatamansi</it> Jones is a formula for treating anxiety-related diseases in the clinic, which is composed of <it>Valeriana jatamansi</it> Rhizoma et Radix, <it>Ziziphi Spinosae</it> Semen, <it>Albiziae</it> Cortex and Junci Medulla. The purpose of this study was to explore the anxiolytic properties of this compound in mice.</p> <p>Methods</p> <p>Male ICR mice were treated with compound <it>Valerianae Jatamansi</it> Jones (1.2 g/kg, 2.4 g/kg, 4.8 g/kg), saline, diazepam (2 mg/kg) orally for 10 days and then exposed to elevated maze-plus (EPM) and light–dark box (LDB). The effects of the compound on spontaneous activity were evaluated by locomotor activity test. We further investigated the mechanism of action underlying the anxiolytic-like effect of compound by pre-treating animals with antagonists of benzodiazepine (flumazenil, 3mg/kg) prior to evaluation using EPM and LDB.</p> <p>Results</p> <p>Compound <it>Valerianae Jatamansi</it> Jones (2.4, 4.8 g/kg, p.o.) significantly increased entries (<it>P</it><0.05) into and time spent (<it>P</it><0.05) on the open arms of the EPM, and number of transitions (<it>P</it><0.05) and time spent (<it>P</it><0.05) in the light compartment of the LDB. However, the anxiolytic-like effects of compound were significantly reduced by pre-treatment with flumazenil (<it>P</it>>0.05). In addition, compound <it>Valerianae Jatamansi</it> Jones treatment didn’t affect the spontaneous activity in mice (<it>P</it>> 0.05).</p> <p>Conclusions</p> <p>The present study supports the hypothesis that compound <it>Valeriana jatamansi</it> Jones exert anxiolytic action but no sedative effects in mice and that this effect might be mediated by benzodiazepine receptors.</p>