四逆散镇静催眠作用的物质基础初步研究

目的初步确认四逆散镇静催眠作用的活性物质,为揭示其镇静催眠作用的机制提供依据。方法采用HPLC方法分析四逆散ig给药后大鼠血中及脑脊液中移行成分,色谱条件为Kromasil C18柱(250 mm×4.6 mm,5μm),体积流量0.8 mL/min,柱温28℃,流动相A为乙腈,流动相B为磷酸水(pH值为3),梯度洗脱(10%～90%A),分析时间为140 min。结果 4种成分辛弗林、芍药苷、柴胡皂苷C、甘草次酸均可使脑脊液中内源性物质5-羟色胺(5-HT)峰面积大于空白脑脊液中峰面积,但作用不如四逆散冻干粉整方强,且混合物组(辛弗林-芍药苷-柴胡皂苷C-甘草次酸)脑脊液中5-HT峰面积是四逆散冻干粉组的3.2倍;在延长戊巴比妥钠所致小鼠睡眠时间方面,辛弗林、芍药苷、柴胡皂苷C、甘草次酸及其混合物和四逆散冻干粉组作用强于对照组,且混合物组优于四逆散冻干粉组。结论通过四逆散冻干粉血和脑脊液中移行成分分析及四逆散冻干粉对戊巴比妥钠所致小鼠睡眠时间影响的药效研究,初步确认了四逆散冻干粉的活性物质是血清中四逆散冻干粉的移行成分辛弗林、芍药苷、柴胡皂苷C及甘草次酸的混合物。

Material basis of Sini Powder with sedative-hypnotic effect

Objective To confirm the active constituents in Sini Powder(freeze-dried powder) with the improving sedative-hypnotic function for the further studies on effective constituents and their mechanism.Methods By using HPLC method and gradient elution,chromatographic conditions were as follows: Kromasil C18 column(250 mm × 4.6 mm,5 μm),flow rate 1.0 mL/min,column temperature 28 ℃;Mobile phase A was acetonitrile and mobile phase B was phosphoric acid water(pH = 3);mobile phase A gradient elution(10%-90% A),and the analysis time was 140 min.Results Synephrine,paeoniflorin,saikosaponin C,and glycyrrhetinic acid could make the peak area of endogenous substance(5-hydroxytryptamine,5-HT) in cerebrospinal fluid larger than that in blank cerebrospinal fluid.But all their effects were less than those of Sini Powder.The increasing peak area of 5-HT in cerebrospinal fluid by the mixture of synephrine-paeoniflorin-saikosaponin C-glycyrrhetinic acid was 3.2 times as much as that of Sini Powder.The effects of synephrine,paeoniflorin,saikosaponin C,glycyrrhetinic acid,their mixture,and Sini Powder on prolonging the sleeping time of mice induced by Pentobarbital sodium were significantly stronger than those in the control group,and the mixture group was better than the Sini Powder group.Conclusion Analysis on transitional composition of Sini Powder in blood and cerebrospinal fluid,and efficacy study on Sini Powder on sleeping time of mice induced by Pentobarbital sodium are carried out,which indicates that the active constituents(synephrine,paeoniflorin,saikosaponin C,and glycyrrhetinic acid) in Sini Powder are confirmed and could improve the sedative function.