Study On The Improvement Of Sleep Active Sites In Suanzaoren

Zizyphus jujube mill.Var.spinous(Bunge)Hu ex h.f.chow)is the dried and mature seed of Rhamnaceae.It was first recorded in Shennong material media and listed as the top grade.It is the primary choice Chinese medicine for insomnia treatment.Modern research is shown that saponins,flavonoids and fatty acids in semen Ziziphi Spinosae have good tranquilizing and tranquilizing effects.Objective: To study the active part of semen Ziziphi Spinosae,which can improve sleep by using the method of pharmacogenetics tracking and modern extraction,separation and purification technology,and to study the material basis and mechanism of pharmacogenetics effect of the active part,so as to provide a theoretical basis for the better development and utilization of semen xiii Spinosae,a traditional Chinese medicine.Methods: 1.Study on the separation and purification of the best active site: firstly,the pharmacokinetics screening of different extracts which have the effect of improving sleep according to the literature reports was carried out based on the number of mouse activities and the central synergy experiment of pentobarbital sodium,from which the best extract of improving sleep was obtained;secondly,the classic drug chemical separation technology combined with modern multi walled carbon nano was used Based on the results of improving sleep activity,the best active part for improving sleep was obtained.2.Study on the basis of effective substances of active parts: the effective components and components in the best active parts were studied by modern analytical techniques such as high performance liquid chromatography and liquid chromatography mass spectrometry.On this basis,according to the effective components and components obtained,the high-performance liquid fingerprint chromatogram was established by using Agilent dad-hplc,and the substance basis of improving sleep efficacy of Suanzaoren was studied by using the established fingerprint chromatogram and serum pharmacochemistry method.3.A study on the basis of effective substances of active parts: the effective components and components in the best active parts were studied by modern analytical techniques such as high performance liquid chromatography and liquid chromatography massspectrometry.On this basis,according to the effective components and components obtained.The high-performance liquid fingerprint chromatogram was established by using Agilent dad-hplc,and the substantial basis of improving sleep efficacy of Suanzaoren was studied by using the established fingerprint chromatogram and serum pharmacochemistry method.Results: 1.The separation and purification of the active part showed that: using the pharmacokinetics tracking method,taking the number of mouse's autonomous activities and the central synergy experiment of pentobarbital sodium as the inspection index,the five extracts designed were screened,and finally extract 2(alcohol extract SZR-E2)was the best extract for Zizyphus spin-offs to improve sleep.Five separation sites were produced by chemical separation method of traditional Chinese medicine.Aiming for the best improvement of sleep activity separation site 4(n-butanol-site SZR-S4),modern multi wall carbon nanotube extraction separation and purification technology was used to obtain the best improvement of sleep active site(refined product 5: n-butanol-eluat SZR-R5).The orthogonal experiment results showed that the best purification process of Zizyphus spinous for improving sleep was as follows: The ratio of the amount of eluate(ml)to MWCNTs(g)was40:1,the concentration of drug solution was 1.5g/ml,and ultrasonic extraction for 30 min.The results of activity verification were: SZR-ACT-MATT>SZR-S4>SZR-E2,which indicated that szr-act-mat was successfully obtained through the above extraction,separation and refining process.2.The study on the basis of the effective substance of the active part shows that:In view of SZR-ACT-MATT,the effective components of SZR-ACT-MATT was analyzed by HPLC.The results showed that the average contents of Jujuboside A,spironolactone,total flavone,total saponin,total alkaloid,total polysaccharide,total phenolic acid,total protein,total fatty acid in szr-act-matt were 0.586,0.184,2.114,6.851,0.985,0.811,0.997,0.031 and5.833,respectively.SZR-ACT-MATT was designated by LC-MS 24 compounds were obtained,including 2 saponins,4 flavonoids and 10 fatty acids.On this basis,by using HPLC and similarity evaluation software of traditional Chinese medicine chromatographic fingerprint,SZR-ACT-MATT fingerprint was established by using multi-point correctionmethod.According to the 14 common peak fingerprint pattern maps generated,9 peaks were identified by comparison of retention time and chromatographic behavior,which were puerarin,ferric acid and spironolactone Rutin,hesperidin,hyaline,quercetin,Jujuboside A and Jujuboside B were all included in the results of SZR-ACT-MATT liquid quality analysis.By utilizing the fingerprints obtained,26 transitional components in rat serum were identified,among which 11 and 15 were metabolites.By comparing the retention time with the control,11 prototypes were identified Seven components were puerarin,ferric acid,spironolactone,rutin,quercetin,Jujuboside A and Jujuboside B.3.The mechanism of action of the active site was studied : The results of morphological observation showed that most of the related neurons in the hypothalamus and hippocampus of the insomnia rats in the positive control group and SZR-ACT-MATT group were recovered compared with those in the model group;the content of Glu and GABA in the hypothalamus of the insomnia rats in the SZR-ACT-MATT group was increased,Glu content was decreased,and GABA content in the hippocampus was increased in the high dose group only-The results of HT and Da showed that SZR-ACT-MATT could increase the 5-HT content of hypothalamus and hippocampus and decrease the Da content at the same time,especially the 5-HT content of hippocampus in the high and medium dose groups of SZR-ACT-MATT;the immunohistochemistry results showed that SZR-ACT-MATT could significantly improve the expression of GABAAR?1 and ?2 positive cells in hypothalamus of insomnia rats There was no significant difference in the expression of GABAAR?1 and ?2positive cells in hippocampus.Based on the above experimental results,it can be confirmed that the mechanism of SZR-ACT-MATT its " SZR-ACT-MATT is based on GABA-A receptor system pathway,which can improve insomnia by regulating the GABA and Glu content of hypothalamus in brain tissue".At the same time,because the high and medium dose of SZR-ACT-MATT can significantly increase the 5-HT content in the hippocampus,we conclude that the mechanism of SZR-ACT-MATT may also be related to the 5-HT receptor in the hippocampus.Conclusion: In this experiment,SZR-ACT-MATT was obtained by modern extraction,separation and purification technology with pharmacogenetics tracking method.On the basis of mass spectrometry analysis,HPLC fingerprint,serum pharmacochemistry,immunohistochemistry and other studies,the pharmacokinetics substances and mechanism of SZR-ACT-MATT were discussed preliminaries,which laid a theoretical foundation for better development and utilization of SZR-ACT-MATT.