Anxiolytic Effects And Its Mechanism Of Passiflora Edulis Sims

Objective:To investigate the process of separating and purificating of total flavonoids from Passiflora edulis and to determine the content of orientin, isoorientin and vitexin in leaves of Passiflora edulis Sims; To investigate the anxiolytic activities of the ethanol extract of Passiflora edulis; To investigate the anxiolytic activities of the ethanol extract of Passiflora edulis fractions, viz ethyl acetate extract (EAE), n-BuOH extract (BE) and aqueous extract (AE) and determine the content of total flavonoids; To examine the effect of the ethanol extract of Passiflora edulis and its fractions, viz ethyl acetate extract (EAE), n-BuOH extract (BE) and aqueous extract (AE) on the mRNA expression of alpha2subunit and alpha3subunitof GABAA receptor.Methods:The content of total flavonoids of Passiflora edulis was analyzed with UV spectrophotometry and the content of orientin, isoorientin and vitexin in leaves of Passiflora edulis Sims was analyzed with HPLC and its condition is that the wavelength was chosen at350nm and the flow rate was1mL/min, The Intertsil C18column and acetonitrile-0.1%phosphoric acid were used as the stationary and mobile phase respectively; Kun ming mice were used as experimental animals in elevated plus-maze (EPM) test to assay the behavioral effects of ethanolic extract (EE) of the aerial part of Passiflora edulis; Kun ming mice were used as experimental animals in elevated plus-maze (EPM) test to assay the behavioral effects of ethanolic extract (EE) of the aerial part of Passiflora edulis fractions, viz.ethyl acetate extract(EAE), n-BuOH extract (BE) and aqueous extract (AE) and the method of AlCl3was used to determine the content of total flavonoids; PCR products were analyzed by RT-PCR and Real-Time PCR method.Results:The static saturated absorbtion capacity of polyamide resin to flavonoids of Passiflora edulis was23.14mg/g. The technical conditions of dynamic absorbtion:the concentration of loading sample is30mL, the purification with distilled water (8times the volume of the polyamide),and then with50%ethanol (12times the volume of the polyamide) was used to elute total flavonoids,and then the eluting velocity was2BV/h, the content of orientin、isoorientin and vitexin were0.957%,0.278%, and0.148%; The aerial part of Passiflora edulis was anxiolytic at400mg/kg/d、800mg/kg/d dose; Ethyl acetate extract(EAE), n-BuOH extract (BE) and aqueous extract (AE) was anxiolytic at100mg/kg/d,200mg/kg/d,300mg/kg/d dose and the content of the total flavonoids were37.43%,21.62%, and6.58%; the ethanol extract of Passiflora edulis can increased the mRNA expression of alpha2subunit and alpha3subunitof GABAA receptor at400mg/kg/d,800mg/kg/d dose, but can not increased the mRNA expression of alpha2subunit and alpha3subunit of GABAA receptor at200mg/kg/d dose and its fractions, viz ethyl acetate extract (EAE), n-BuOH extract (BE) and aqueous extract (AE) can increase the mRNA expression of alpha2subunit and alpha3subunitof GABAA receptor at300mg/kg/d dose.Conclusion:The total flavonoids of Passiflora edulis is able to be easily separated and purified by polyamide resin under the optimum conditions above; The ethanolic extract (EE) of the aerial part of Passiflora edulis has anxiolytic efficacy; ethyl acetate extract (EAE), n-BuOH extract (BE) and aqueous extract (AE) has anxiolytic efficacy and the anxiolytic effect may be associated with the content of total flavonoids; The aerial part of Passiflora edulis is to increase the mRNA expression of alpha2subunit and alpha3subunitof GABAA receptor to play an anxiolytic effect.