Study On Separation,purification And Biological Activity Of Zizyphi Spinosae Semen Saponins

Ziziphus jujuba Mill.var.spinosa（Bunge）Hu ex H.F.Chow,belonging to genus Rhamnaceae is both traditional Chinese herbal medicine and medicinal and food homologous variety.Zizyphi Spinosae Semen is the seed of Ziziphus jujuba.Modern pharmacological studies have shown that Zizyphi Spinosae Semen has good sedative and hypnosis activity,and the main active ingredient is Zizyphi Spinosae Semen saponins.At present,there are many reports on the sedative hypnosis of Zizyphi Spinosae Semen saponins,but little information is available about its other activities.In this paper,Zizyphi Spinosae Semen saponin was used as the research object,and its components are identified by spectroscopy,and its antioxidant,anti-tumor,hypoglycemic,anti-aging,and sedative and hypnotic activities were carried out.（1）After heating reflux extraction,organic solvent fractionation extraction and macroporous resin purification,the primary saponins（SZR Ⅰ）and secondary saponins（SZR Ⅱ）of Zizyphi Spinosae Semen were obtained.The content of SZR Ⅰ saponins was 52.1%and SZR Ⅱ saponins was 72.1%.Using fourier infrared transform and LC-MS spectrometry analysis,the results show that there are differences in the composition of SZR Ⅰ and SZR Ⅱ.SZR Ⅰ contains betulinic acid,jujuboside A,jujuboside B,spinosin,and SZR Ⅱ contains jujuboside A,jujuboside B,spinosin,jujuboside B₁.（2）The antioxidant and anti-tumor activitives of SZR Ⅰ and SZR Ⅱ are explored.SZR Ⅰ and SZR Ⅱ have certain antioxidant effect,and the antioxidant effect of SZR Ⅱ was better than that of SZR Ⅰ;SZR Ⅰ and SZR Ⅱ had almost no antitumor effect.（3）The results of hypoglycemic experiment show that SZR Ⅰ and SZR Ⅱ have a strong inhibitory effect onα-glucosidase andα-amylase.Meanwhile,SZR Ⅰ and SZR Ⅱ can improve the insulin resistance of Hep G2 cells,which may be related to jujuboside A and spinosin.Further investigation showed that SZR Ⅰ and SZR Ⅱ can increase the activity of superoxide dismutase（SOD）,reduce the content of malondialdehyde（MDA）,and also increase the activity of hexokinase（HK）and pyruvate kinase（PK）.（4）The anti-photoaging effects of SZR Ⅰ and SZR Ⅱ were investigated by Hacat.The results showed that SZR Ⅰ and SZR Ⅱ can promote the proliferation of UVB-damaged Hacat cells.Moreover,jujuboside A,jujube B and spinosine also shows similar effects.In-depth studies have found that they can reduce the level of reactive oxygen species（ROS）and the release of human metal matrix protease（MMP-1）,increase catalase（CAT）and glutathione peroxidase（GSH-PX）activity.Experiments with C.elegans as a model show that SZR Ⅰ and SZR Ⅱ can delay the senescence of nematodes caused by heat and oxidative stress,which may be related to their ability to improve the activity of SOD and CAT.（5）SZR Ⅰ and SZR Ⅱ have protective effects on corticosterone-induced SH-SY5Y cells damage.The insomnia model of ICR mice induced by chlorobenzoic acid（PCPA）was used to explore the sedative and hypnotic effects of SZR Ⅰ and SZR Ⅱ.Studies have shown that SZR Ⅰ and SZR Ⅱ can increase the sleep time of insomnia mice and reduce the sleep latency time.Further experiments show that SZR Ⅰ and SZR Ⅱ can increase the contents of5-hydroxytryptamine（5-HT）,dopamine（DA）,norepinephrine（NE）,interleukin IL-1βand carbon monoxide（NO）in the plasma of insomniac mice,and decrease the contents of glutamate（Glu）and interleukin IL-6.RT-q PCR and western blot results show that SZR Ⅰ and SZR Ⅱ may play sedative and hypnosis effect by regulating 5-HT_1A,5-HT_2A,GABA_ARα2,GABA_ARα3,IL-1β,IL-6 m RNA expression and protein expression.