| Objective: In this paper,the Sugemule-4 decoction was used as the research object,and the basic components of its pharmacodynamics were studied.The pharmacodynamics of Sugemule-4 decoction in the treatment of insomnia and its related neurotransmitters were studied through animal experiments.The influence of quality level provides a reliable theoretical basis for the development of Sugemule-4 decoction.Methods:(1)The Mongolian drug Sugemule-4 decoction was extracted by hexane,ethyl acetate and n-butanol system.The hexane and ethyl acetate were detected by GC-MS.Peaks and their corresponding compounds were identified through database index.(2)Institute of Cancer Research(ICR)mice were selected and divided randomly into 5 groups.Through the experiments of measuring the frequency of mice' autonomic activities,the hypnotic effect of pentobarbital sodium and the subthreshold hypnotic effect of pentobarbital sodium,the number of autonomic activities,sleep latency,sleep time and sleep rate of mice in the 5groups were compared.After the last administration and recording,the mice were anesthetized and the eyeballs were removed for blood biochemical detection.(3)ICR mice were randomly divided into blank group,model group,Sugemule-4 decoction high,medium and low dose groups,and Anshen Bunao liquid control group,10 mice in each group.Duplicate insomnia model mice by means of p-chlorophenylalanine(PCPA)abdominal injection.The mice were injected with PCPA for 2 days.After the model was successfully duplicated,the mice were intragastrically administered every day at 8:00 am on the third day.The Anshen Bunao liquid control group was intragastrically administered at a concentration of 0.13ml/10 g.The high,medium and low dose groups of Sugemule-4 decoction were administered by 52mg/10 g,26mg/10 g,13mg/10g(calculated as raw drug content).The blankcontrol group and the model control group were intragastrically administered with the same volume of distilled water.Before administered,the mice were fasted for 12 h,once a day,and continuously administered for 7 days.After the last administration for 24 hours,the mice were treated with decervical treatment to obtain brain tissue.The effects of Sugemule-4decoction on the six neurotransmitters: Serotonin(5-HT),norepinephrine(NE),dopamine(DA),gamma-aminobutyric acid(GABA),glutamic acid(GLU),glycine(GLY)in mouse brain tissue were observed by LC-MS/MS.Results: A total of 59 components were isolated from the hexane fraction,43 components were isolated from the ethyl acetate fraction,and most of them were identified.Its chemical composition in hexane and ethyl acetate extracts consists mainly of terpenoids,triterpenoids,phenylpropionic acids,alkaloids,steroids,aliphatic compounds,aromatic acids,organic acids,aldehydes,esters,alcohols,hydrocarbons and many derivatives of benzene derivatives.These ingredients are known to be paeonol,piperine,etc.may be related to the anti-insomnia effect of the drug.The number of autonomic activities in Sugemule-4 each dose group compared with the normal control group,the differences were statistically significant((49)<0.05).The sleep latency of Sugemule-4 high dose group and Anshen Bunao Liquid control group were significantly shorter than that of normal control group,the differences were statistically significant((49)<0.05).There was no significant difference in sleep latency of between Anshen Bunao Liquid control group and normal control group((49)>0.05).The sleep time of Anshen Bunao Liquid control group and Sugemule-4 each dose group were significantly longer than that of normal control group,the differences were statistically significant((49)<0.05).The sleep rate of mice in Anshen Bunao Liquid control group,and Sugemule-4 each dose group were significantly better than that of normal control group,the differences were statistically significant((49)<0.05).The alanine aminotransferase(ALT),aspartate aminotransferase(AST),AST/ALT,blood glucose(GLU),glutamyltranspeptidase(GGT),blood uric acid(Blood uric acid,UA-G),serum creatinine(CRE-G),total bilirubin(TBIL),direct bilirubin(DBIL),indirect bilirubin(IDBIL)index in each dose group of Sugemule-4 were no significantly different from that in the normal control group((49)>0.05).The total cholesterol(Cholesterol,CHOL)index in the low dose group of Sugemule-4 was significantly different from that inthe normal control group and the Sugemule-4 medium dose group(P<0.05).The contents of5-HT,GABA,GLY and NE in the brain of Sugemule-4 high-dose group were significantly higher than those in the PCPA model group((49)<0.05),and the contents of DA and GLU were significantly decreased((49)<0.05).The content of GABA in the brain tissue of the mice in the Sugemule-4 medium dose group was significantly higher than that in the PCPA model group((49)<0.05).The content of 5-HT and GLY was higher than that in the PCPA model group but did not reach a significant level((49)>0.05).The content of GLU is significantly reduced((49)<0.05),The content of DA and NE decreased significantly but did not reach a significant level((49)>0.05);The content of GLU in the brain tissue of the mice in the low dose group of Sugemule-4 was significantly higher than that in the PCPA model group((49)<0.05),but the content of DA and NE decreased but did not reach a significant level((49)>0.05).The contents of 5-HT,GABA,and GLY were increased compared with the PCPA model group but did not reach a significant level((49)>0.05).Conclusion: The GC-MS method was used to compare the low polarity chemical composition of Mongolian medicine Sugemule-4 decoction for the first time.The extract contained higher concentrations of piperine related to hypnosis.It also contains other active ingredients such as paeonol and linalool.Sugemule-4 decoction has a certain degree of sedative and hypnotic effects on mice,which can promote sleep,prolong sleep time.The biochemical tests of mice showed that there was no significant difference in liver and kidney function between the experimental mice in the experimental group and the normal control mice.Sugemule-4 decoction can significantly increase the levels of 5-HT,GABA,GLY and NE in the brain of PCPA-induced insomnia mice,reaching normal levels,among which the neurotransmitters 5-HT,GABA and GLY,with the increase of the contents,the inhibition effect increases;and can reduce the contents of DA and GLU in the brain of insomnia mice,and reduce the content of the awakening neurotransmitter,so that the sleep condition of PCPA-induced insomnia mice is improved. |
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