The Study On Mechanism Of Sinisan Improving Sleep

In the past,a large number of clinical and experiments showed thattraditional compound Chinese medicine SNS has significant improvements insleep,but its mechanism in improving sleep is unknown.In order to clarifythe mechanism of SNS improve sleep,the experiment begin with sodiumpentobarbital collaborative experiment that evolve the research aboutdose-response relationship and time-concentration of SNS freeze-driedpowder.We carried out multiple-targets' mechanism research of SNSfreeze-dried powder improve sleep,based on the GABAergic nervous system,serotonergic nervous system,NO signaling pathway,theory of sleep mediatedby SP,IL-1βand TNF-α,used a series of research methods and technologysuch as sodium pentobarbital collaborative experiment,brain microdialysis,HPLC-EC technique,RT-PCR tech nology and enzyme linked immunosorbentassay.The results:1.The research about dose-response relationship and time-concentration ofSNS freeze-dried powder improve sleep.In the dose-response relationship study established five dose(312.5,625,1250,2500,5000mg/kg,i.g.,7 days),the best dose is 1250mg/kg,which registeras extend pentobarbital sodium-induced sleep time in mice and shorten theincubation period,exceed or lower than this dose,both have no synergy withpentobarbital sodium.In the time-concentration study(3 days,5 days,7 days,9 days),time to onset of SNS freeze-dried powder (1250 mg/kg) iscontinuous intragastric administration 7 days.Continuous intragastricadministration 9 days,the effect of synergy with pentobarbital sodium getmaximize.2.The mechanism research based on SNS improves sleep mediated byGABAergic nervous system.Flumazenil has obvious antagonism against the enhancement of sleep inmice of pentobarbital sodium-induced by diazepam (2.5mg/kg,i.g.) and SNS freeze-dried powder (1250mg/kg,9days,i.g.),Theresults showed that SNSfreeze-dried powder improve sleep through the BDZ receptor-mediated.On the bases,use RT-PCR technology observes the expression of mRNAof GABAA receptor al and a5 subunit which acted by SNS freeze-dried powder.Experimental results show that,SNS freeze-dried powder raise the expressionof mRNA of GABA_A receptor al and a5 subunit in mice cerebral cortex,itsrole is similar to diazepam (2.5mg/kg,i.g.),however,has different intensity.SNS freeze-dried powder raise focus on the a5 subunit as well as diazepamraise focus on the al subunit.3.The mechanism research based on SNS improves sleep mediated by NO signalingpathway.SNS freeze-dried powder (1250 mg/kg,9 days,i.g.) can significantlyincreased NO content in the whole brain of mice and enhance the vitality ofNOS.Subthreshold doses of SNS freeze-dried powder (312.5 mg/kg,9 days,i.g.) combined application of L-Arg (125mg/kg,i.p.) can significantly prolongsodium pentobarbital-induced sleep time in mice,which showing a certainsynergy.However,threshold dose of SNS freeze-dried powder (1250mg/kg,9 days,i.g.) combined application of L-Arg (500mg/kg,i.p.) can significantlyshorten sodium pentobarbital-induced sleep time in mice.Given antagonistL-NAME (80 mg/kg,ip)can obviously antagonism of the hypnotic effects ofSNS.Based on the above findings show that the improvement in sleep of SNSis by regulating the content of NO.4.The mechanism research based on SNS improves sleep mediated byserotonergic nervous system.Subthreshold doses of SNS freeze-dried powder (312.5 mg/kg,9days,i.g.)combined application of 5-HT precursor 5-HTP(25mg/kg,ip) can significantlyprolong sodium pentobarbital-induced sleep time in mice,which showing acertain synergy.Subcutaneous injection of PCPAcan significantly shortensodium pentobarbital-induced sleep time in mice,for such mice,the thresholddose SNS has obvious antagonism.The action of SNS prolongs sodium pentobarbital-induced sleep time in mice can interdiction by PCPA.The abovefindings indicate the role of SNS improve sleep mainly depends on theexistence of 5-HT.Give SNS combined p-MPPI or Ritanserin to experiment animal of mice.p-MPPI (0.05mg/kg,i.p.) can obviously antagonism of the effect of SNS(1250mg/kg,9days,i.g.) prolong sodium pentobarbital-induced sleep time in mice,there was no significant difference between merger Ritanserin (8 mg/kg,ip) or givenalone SNS freeze-dried powder (1250 mg/kg,9 days,ig).The research results showthat the extension of SNS prolongs sodium pentobarbital-induced sleep time in micehas nothing to do with the 5-HT₂ receptor but the 5-HT_1A receptor.Using the method of vivo microdialvsis and electroencephalogram/electromyogram (EEG/EMG)synchronous recording in rats,observed the impactof SNS act on release of the contents of 5-HIAA in raphe nucleus of brain in run-freerats.The results show that after intragastric administration of SNS can besignificantly reduced the content of 5-HIAA in raphe nuclei during awark andNREMS period.5.The mechanism research based on SNS improves sleep mediated by SP.Use enzyme linked immunosorbent assay,observed the influence of SPcontent in whole brain of mice acted by SNS.The experimental results showthat SNS freeze-dried powder (1250mg/kg,9days,ig) can significantly decreasedthe SP content in whole brain of mice.6.The mechanism research based on SNS improves sleep mediated by cytokine.Use enzyme linked immunosorbent assay,observed the influence of IL-1βand TNF-αcontent in whole brain of mice actd bv SNS.The experimentalresults show that SNS freeze-dried powder (1250mg/kg,9 days,ig) cansignificantly decreased the IL-1βand TNF-αcontent in whole brain of mice.Summary:Based on the above results show that SNS improves sleep throughmultiple-targets' mechanism.