Preliminary Study On Antidepressant Effect Of Total Alkaloids From Uncaria And Its Serum Pharmacochemistry

Depression is a psychiatric disorder characterized by low mood,and is graduall y becoming one of the most important public health problems.At present,in clinica l,western medicine is given priority to be used to treat the disease,but western m edicine treatment is easy to relapse and toxic side effects and other issues,which g reatly affected the clinical efficacy.Traditional Chinese medicine has the characterist ics of stability,lasting effect,and less adverse reaction,has a long history in terms of the prevention of depression,is now becoming a hot research at home and abro ad.Uncaria rhynchophylla.(Miq.)Miq.exHvail.,the main variety of Uncaria,is a traditional Chinese medicine,with sedative,anticonvulsant,anxiolytic,neuroprotective and anti-Alzheimer's disease and so on.In recent years,studies have shown that U ncaria macrophylla Wall alkaloids,Uncaria lanosa Wallich var.Appendiculata Ridsd ethanol extract and isorhynchophylline have antidepressant effect,but the antidepres sant research of Uncaria rhynchophylla.(Miq.)Miq.exHvail is rarely reported.Indole alkaloids are the main effective components,according to whether the C-2 oxidated which can be divided into the type of oxidation indole alkaloids and in dole alkaloids.The earlier research showed that different regions of Uncaria rhynch ophylla.(Miq.)Miq.exHvail.can be divided into two large categories including S1 main containing oxidationindole alkaloids and S2 mainly containing the indole alka loids.Therefore,based on the previous experiments and the extensive pharmacokinet ic activity of Uncaria,the depressive model mice were prepared by CRS to explore the antidepressant activity of total alkaloids from Uncaria rhynchophylla.(Miq.)Mi q.exHvail,so as to provide the basis for the antidepressant effect of the total alkal oids from Uncaria rhynchophylla,and to observe and compare the pharmacological effects of the two groups of S1 and S2 which components are different,and to deepen the previous research results.Therefore,CRS was used to prepare depressive mo del mice to observe and compare the antidepressant effects of total alkaloids of S1 and S2 on CRS mice.Clarify the effectiveness of Chinese medicine in addition to discover its effectiveness and mechanism of action,but also should be aware of what material works.Compared with the extensive pharmacological activity of Uncaria alkaloids,its in vivo and metabolic processes are not clear.Traditional Chinese medicine serum pharmacochemistry is based on traditional drug chemistry research methods,with a variety of modern analysis techniques,analysis of oral administration of serum after the migration of components(including the prototype components and metabolites),and to study its relevance with the efficacy,Thus a clear subject of traditional Chinese medicine efficacy material basis.Therefore,on the basis of an comprehensive and accurate grasp for the vitro composition of Uncaria rhynchophylla.(Miq.)Miq.exHvail.Extract,and according to the theory and methods of serum pharmacochemistry of Traditional Chinese Medicine(TCM),the preliminary study on the transitional constituents to blood of Uncaria alkaloids extract of S1 and S2 in rats was be carried out in this study.The main experimental content are as follows: Objective:To study the antidepressant effect of(total alkaloids from Uncaria,TAU)and its possible mechanism,and to study the blood components in TAU of S1,S2 and t o further explore its pharmacodynamic material basis.Methods:1?The mice were restrained for 8 h daily in a special centrifuge tube for 10 consecutive days to establish the mode of depressive mice.During CRS,all mice were intragastrically administrated TAU of S1 and S2(80,40 and 20 mg/kg)and fluoxetine(FLU,10 mg/kg).The behavioral changes were observed by spontaneous activity test,forced swim test and sucrose preference test to evaluate antidepressant effect of the drugs.The ELISA method was used to detect the levels of the serum corticosterone(CORT),TNF-? and NO;To establish an HPLC-FLD method for the investigation of monoamine neurotransmitters(NE,E,Tyr,DA,5-HIAA,5-HT and Trp)in mice brain,and this method was used to analyze the contents differences of monoamine neurotransmitters in different groups.2?The difference of components of Uncaria alkaloids extract of S1,S2,drug-c ontaining serum and blank serum were compared with UFLC-IT-TOF-MS technology to determine its blood components.Results:1?Compared with the control group,after restrained in a special centrifuge tube for 10 consecutive days,the mice,s spontaneous activity,body weight and sucrose preference were significantly decreased,and the mice,s immobility period were significantly prolonged,Futhermore,the levels of mice,s CORT,TNF-? and NO in serum and its contents of NE,E,Tyr,DA,5-HIAA,5-HT and Trp were significantly decreased;Compared with the model group,the TAU of S1 and S2 both could significantly increase body weight,shorten immobility period and enhance sucrose preference of the depression model mice.Futhermore,the TAU of S1 and S2 both could significantly decrease the contents of CORT,TNF-? and NO in depressive mice,and the TAU of S1(80,40 and 20 mg/kg)all could significantly increase the contents of 5-HIAA,5-HT and Trp,but had no significant effect on the contents of NE,E,Tyr and DA,while the TAU of S2(80,40 and 20 mg/kg)all could significantly increase the contents of NE,E,Tyr,DA,5-HIAA,5-HT and Trp.2?Twenty six constituents absorbed into rat blood in Uncaria alkaloids extract of S1 containg seven prototype components(C,IC,R,IR,Isorhynchophyllic acid,DCA and DCAB)and nineteen metabolities,and twenty three constituents absorbed into r at blood in Uncaria alkaloids extract of S2 containg seven prototype components(G M,HT,HS,C,IC,R,and IR)and sixteen metabolities were identified by co-chromatogr aphy of samples and comparison of the retention time,characteristic molecular ions and fragment ions with those of the authentic stands or tentatively identified by MS/MS data based on the related literature.Conclusion:1?The total alkaloids from Uncaria rhynchophylla.(Miq.)Miq.exHvail.has obvious antidepressant activity in CRS mice,its mechanism may be related to regulati on of hypothalamus-pituitary-adrenal(HPA)axis function,reduce serum TNF-? and NO levels and increase the contents of monoamine neurotransmitters in the mice br ain.However,each of the drugs on the role of brain tissue in mice with the target point of the neurotransmitter has a certain focus.The TAU of S1 mainly containin g oxidized indole alkaloids have antidepressant effects by increasing the contents of5-HIAA,5-HT and Trp in mice brain tissue,but the effects of NE,E,Tyr and D A were not significant.The TAU of S2 mainly containing non-oxidized indole alkal oids have antidepressant effects by increasing the contents of 5-HIAA,5-HT,Trp,N E,E,Tyr and DA.2?The serum pharmacochemistry study of Uncaria rhynchophylla.(Miq.)Miq.exHvail.alkaloids extract of S1,S2 showed that the oxidation indole alkaloids mainly by hydrolysis,oxidation,glucuronidation and hydroxylation,methylation and water adduct as metabolic pathway and non-oxidation indole alkaloids are mainly demethylated,dehydrogenation and water adduct for metabolic pathway,and the compounds absorbed into blood and its metabolites may well be the effective constituents,which provides scientific basis for clear its pharmacodynamic material basis and reference for the rational use of these two types of Uncaria.