酸枣仁总皂苷提取工艺优化及其改善睡眠作用机制研究

Study on the optimization of extraction process of total saponins of Semen ziziphi spinosae and its mechanism of improving sleep

目的优化酸枣仁总皂苷的提取工艺,并研究其改善睡眠作用。方法本研究采用单因素和响应面实验优化高压脉冲电场法提取酸枣仁总皂苷的工艺参数。同时采用随机法把小鼠分为正常对照组、模型对照组、阳性对照组和酸枣仁总皂苷低、中、高剂量组。除正常对照组外,其余5组灌胃对氯苯丙氨酸(parachlorophenylalanine,PCPA)溶液构建失眠小鼠模型。记录各组小鼠3 min内的水平、垂直和修饰活动得分,测定小鼠神经递质中5-羟色胺(5-hydroxytryptamine,5-HT)、去肾上腺素(noradrenaline,NE)、多巴胺(dopamine,DA)、谷氨酸(glutamic acid,Glu)和γ-氨基丁酸(γ-aminobutyric acid,GABA)的含量。结果最佳提取工艺条件为:乙醇浓度71%、电场强度20 kV/cm、脉冲数10,此条件下酸枣仁总皂苷得率为7.18%。改善睡眠实验结果表明:酸枣仁总皂苷能降低PCPA诱导失眠模型小鼠的水平、垂直和修饰得分,升高失眠模型小鼠神经递质中5-HT和DA的含量,降低NE的含量,同时降低Glu含量和Glu/GABA的比值,提高GABA含量,表明其对PCPA诱导失眠模型小鼠有良好的改善睡眠作用。结论该研究获得了酸枣仁总皂苷的最佳提取工艺,得到的酸枣仁总皂苷对PCPA诱导失眠模型小鼠有良好的改善睡眠作用。

Objective To optimize the extraction process of total saponins of Semen ziziphi spinosae and study its effect on improving sleep.Methods The single factor and response surface experiments were used to optimize the process parameters for extracting total saponins from jujube kernel using high-voltage pulse electric field method.At the same time,mice were randomly divided into normal control group,model control group,positive control group,and low,medium,and high dose groups of total saponins of Semen ziziphi spinosae.Except for the normal control group,the other 5 groups were gavaged with parachlorophenylalanine(PCPA)solution to construct an insomnia mouse model.The horizontal,vertical,and modified activity scores of each group of mice within 3 minutes were recorded,and the levels of 5-hydroxytryptamine(5-HT),norepinephrine(NE),dopamine(DA),glutamate(Glu),and the content ofγ-aminobutyric acid(GABA)were determined.Results The optimum extraction conditions were:The ethanol concentration was 71%,electric field intensity was 20 kV/cm,and pulse number was 10.Under these conditions,the yield of total saponins of Semen ziziphi spinosae was 7.18%.The results of the sleep improvement experiment showed that the total saponins of Semen ziziphi spinosae could reduce the horizontal,vertical,and modification scores of PCPA induced insomnia model mice,increase the content of 5-HT and DA in the neurotransmitters of insomnia model mice,reduce the content of NE,while reducing the Glu content and Glu/GABA ratio,and increase the GABA content,indicating that it had a good sleep improvement effect on PCPA induced insomnia model mice.Conclusion The best extraction process of total saponins of Semen ziziphi spinosae is obtained,and the total saponins of Semen ziziphi spinosae have a good sleep improvement effect on PCPA induced insomnia model mice.