| Objective:Stress,also known as stress response,are adaptative nonspecific responses of whole body,when suffered all kinds of environment society and psychological factors.At the beginning,stressor induces the disorder of homeostasis inside body.But when the stressor affects body continuously,diseases occur.Long－time chronic stress easily induces abnormal behaviour of anxiety,depression in rats,companying with related endocrine changes.In earlier stage,our team has researched liver depression and spleen deficiency syndro me of rats induced by 21 days chronic immobilization stress for many years.So far,rat depression molding medthods,including 21 days chronic unpredictable mild stress,21 days chronic unpredictable stress and 21 days chronic immobilization stress,have been universally acknowledged.However,how to make rat anxiety model with liver depressbn and spleen deficiency syndrome is still uncertain.Our team has compared behavior,related protein and gene of brain from rats of different molding durations,and found that 14 days chronic immobilization stress was able to establish anxiety model of liver depression and spleen deficiency syndrome.In order to identify the model,the effects of Chinese medicine Xiaoyaosan on rat model,and explore the mechanism of Xiaoyaosan treating anxiety with liver depression and spleen deficiency syndrome,based on previous foundation and related references,behavioristics and included molecular biological indexes in JNK/MAPK signaling pathway were observed and tested.As the same time,fluoxetine ’was used as positive nedicine to evaluate effects of Xiaoyaosan,and SP600125,inhibitor of JNK/MAPK signaling pathway,was applied to research the mechanism.Methods:(1)36 SD rats were randomly divided 4 groups.Beside normal groiup,the other was given continuous 14 days chronic immobilization stress,3h/d.Model group,Xiaoyaosan group and fluoxetine group were respectively gavaged corresponding drugs.Body weight and general status of rats were observed in o,7,15 days,and behavioristics,such as open-field test,glucose consumption test,elevated plus-maze test,and so on,were observed and evaluated.(2)After 4 group rats were executed,Western blot and qRT-PCR methods were used to detect P-JNK,JNK,P-c-Jun contents in hippocampus.(3)40 SD rats were randomly 5 groups,8 rats each group.Sham operation group and SP600125 group were given bilateral ventricle cathetering operatioa After 1w recovery,beside normal group,the other four groups were given chronic restrain stress,continuous 14 days,3h/d.Each group was given corresponding drug.After molding,general status,body weight,behavioristics indexes were evaluated.(4)JNK,C-Jun,Cyt-C protein and gene were tested by Elisa,Western blot and qRT-PCR methods.Results:(1)After 14 molding,normal group,Xiaoyansan group and fluoxetine group rats are in good condition,bright and tidy hair,active,calm,moderate stool,while model group rats look like frail,weak,anxiety,messy hair,sloppy stooL Body weight of model group is lower than the other groups,and the difference is significant(P<0.05).There are no differences among four groups in glucose consunption and open-field test In elevated plus-maze test,total distaince,open arms entrance times and detention durations in model group are shortened than the other groups(P<0.05,P,0.01).In novelty suppressed feeding test,feeding time of model group rats is obvious longer than the other groups(p<0.05).(2)There are statistical differences between model group and the other 3 groups in P-JNK,JNK,P-c-Jun protein expressions and JNK,c-Jun mRNA in hippocampus(P<0.05,P<0.01).(3)Sham operation grovp and model group rats in this experiment have similar behavioral performance with model rats in experiment 1(P<0.05,P<0.01),while Xiaosansan group and SP600125 group have similar manifestations with normal group.(4)Elisa results show P-JNK,P-c-Jun protein expressions of hippocampus in model and sham operation group are higher than the other groups(P<0.05,P<0.01).Western blot method indicates P-JNK,JNK,P-c-Jun protein expressions in model and sham operation groups are higher than normal group(P<0.05,P<0.01),and there are no statistical differeces among normal group,Xiaoyaosan group and SP600125 group.qRT-PCR method indicates JNK,c-Jun mRNA egressions in model and sham operation groups are higher than the other 3 groups(P<0.01).What’s more,there is no difference of Cyt-c protein and mRNA expression among 5 groups.Conclusion:(1)Xiaoyaosan as one of classical prescriptions,has certain effects on anxiety model rats with liver depression and spleen deficiency syndrome.(2)14 days chronic immobilization stress can induce rat anxiety-like behaviors,and successfully establish rat model of anxiety.(3)Antianxiety effects of Xiaoyaosan on model rats of liver depression and spleen deficiency syndrome are related with JNK/MAPK singling pathway.(4)The mechanism of Xiaoyao san action has nothing to do with Cyt-c. |
PDF Full Text Request