Effects Of An Shen Jie Yu Decoction On Behavior As Well As Cytoskeleton Protein And HPA Axis Of The Hippocampus In Rats With CUMS

Objectives The depression model of rats was established by chronic unpredicted mild stress(CUMS).The effects of An Shen Jie Yu Decoction(ASJYD)on behavior as well as microtubule-associated protein and hypothalamic-pituitary-adrenal(HPA)axis of the hippocampus in rats with CUMS were detected to clarify the pathogenesis of depression and provide some experimental basis for treatment.Methods Sixty male SD rats(180～220g)were randomly divided into control group(Control),depression model group(CUMS),and An Shen Jie Yu Decoction treatment group(ASJYD)(n = 20),respectively.The chronic unpredicted mild stress(CUMS)was used to establish a model of depression in rats through a classic modeling method.Rats in the control group were raised normally without special treatment.After CUMS,the treatment group was administered with a dose of 5.5 g/kg of ASJYD to depressed rats by intragastric administration.Rats were sacrificed after continuous intragastric administration once a day for 14 days,Behavioral testing of each group at corresponding time points.The behavior of the control group,the CUMS group,and the treatment group was observed with the sucrose preference test,open field test,and Morris water maze experiments.HE staining was used to observe the changes of hippocampal tissue structure.Transmission electron microscopy was used to observe the ultrastructure of hippocampal neuron cells of each group of rats.The location expression of phosphorylation-Tau(p-Tau)in the hippocampus of each group was observed by immunofluorescence.The expression level of tubulin p-Tau was detected by immunoblotting.ELISA was used to detect the changes of adrenocorticotropic hormone and(ACTH)and croticotropin-releasing facto(CRF)in each group of rats.The data were analyzed by SPSS19.0,and the difference was statistically significant at P <0.05.Results 1 Sucrose preference experiment: There was no significant change in tap water consumption in each group.The amount of sucrose consumption and the percentageof sucrose preference in the model group were significantly lower than those in the control group and the treatment group(P < 0.01).2 Open field experiment: The walking distance,central-lattice time,standing frequency,and grooming behavior in the CUMS group were significantly lower than those in both the control group and the ASJYD treatment group(P<0.01).3 Morris water maze experiment: The average escape latent period in both the control group and the ASJYD treatment group was lower than that in the CUMS group(P<0.01).4 Nissl staining: The hippocampal tissue structure of each group changed significantly.The results showed that the hippocampal cells of the control group and the ASJYD treatment group were neatly arranged,with a high density and a large number,and the nucleus was normal.The hippocampal cells in the CUMS group were irregularly arranged,with a low density and a small number,and the nucleus was incomplete.5 Transmission electron microscopy observation: The results showed that the nuclear membrane of hippocampal neurons in the control group and the ASJYD treatment group was relatively complete,and the ultrastructure was normal.The nucleus of hippocampal neurons in the CUMS group atrophied.It was found that the endoplasmic reticulum swelled,organelles are relatively reduced and structures were destroyed,and vesicles appeared in the cytoplasm.6 Immunofluorescence results: p-Tau was observed using red fluorescence excited by Alexa Fluor 594,and the results showed that p-Tau was expressed in the cytoplasm of hippocampal neurons.7 Western Blot test results: The protein expression of p-Tau protein in the model group was significantly lower than that in the control group after CUMS(P<0.01),and it was significantly increased after treatment with ASJYD compared with the CUMS group(P<0.01).8 The ELISA test results showed that the CRF content and serum ACTH level in the hypothalamus of the CUMS group were significantly higher than those of the control group,and there was a significant improvement after treatment with ANJYD(P<0.01).Conclusions 1 The structure of rat hippocampal neurons changed after CUMS.2 The expression level of cytoskeleton tubulin p-Tau was significantly reduced,suggesting that it may be related to changes in hippocampal cytoskeleton.3 ASJYD has antidepressant effect on CUMS rats,which can regulate the changes of hypothalamus CRF and serum ACTH,the mechanism of action may be closely related to the regulation of the HPA axis.Figure11;Table6;Reference 102...