Effect Of Tonify Deficiency And Remove Blood Stasis Therapeutic Method On ERα,ERβ,BDNF,TrkB Expression In Hippocampus Of Postpartum Depression Model Rats

Objective:We intend to intervene the postmartum depression rat model by Chinese medicine chanshu granules (shenguiren mixture), Then observe the changes of behavior in rats. detect serum estrogen (E2) levels, estrogen receptor (ERa, ERP) and brain neurotrophic factor (BDNF), tyrosine kinase receptor (TrkB) mRNA and proteinexpression in hippocampus of PPD model rats.To explorethe correlation between the method of Tonify deficiency and remove blood stasis, estrogen, BDNF and postpartum depression. And then Reveal the mechanism and scientific connotation of tonify deficiency and remove blood stasis therapeutic method treat postpartum depressionMethods:1. A total of 120 qualified by open field test screening of mature female SD rats. Randomly divided into normal control group (Ctl) 16, The other rats were prepared with PPD models. PPD models were received continuous subcut-aneous injection of estrogen and progesterone after castration. The depressionstatus of PPD model rats were evaluated after modeling by using sugar consumption experiment,Open Field Test and Forced Swimming Test.According to the results of the open field test, PPD model rats were randomly divided into 6 groups,16 rats in each group:①The shenguiren mixture of high dose group (S-H group),② the reference to dose group (S-M group),③kernel mixture in the shenguiren mixture of low dose (S-L group), ④the estradiol benzoate group (EB group),⑤and sertraline hydrochloride group (Ser group),⑥the model group (Mdl group).2.To start the drug intervention after categorized. S-H, S-M, S-L group were intragastrically administered with high, medium and low dose of Chinese medicine shenguiren mixture. Ser group received intragastric administration of positive control drug sertraline hydrochloride.Ctl group, Mdl group were given equal volume of pure water. EB group use positive control drug estradiol benzoate subcutaneous injection. Drugs were chronically administered to animals for 4 weeks.All of the rats were observed with behaviors tests for 2 weeks and 4 weeks.3.After 4 weeks of administration,correlated indexes were detected as follow: ①observing serum E2 level by RIA.②checking expression of ERa、ERβ、BDNF、 TrkB mRNA in hippocampus in brain through RT-PCR.③measuring protein expression of ERa、ERβ in hippocampus by the method of immunohistochemical. ④measuring protein expression of BDNF, TrKB in hippocampus of brainby Western blotting (WB)Results:1.behavior index:After the modeling, compared with Ctl group, PPD rats sugar consumption significantly reduced, open field test score level and verticalscore decreased significantly, real time increased significantly in the forced swi-mming test.After drug intervention.Except Ser group, other groups for 2 weeks,4 weeks of sugar consumption were increased significantly compare with Mdl group. Each treatment group open field level scores were higher than that in Mdl group (P<0.01).Vertical scores of Each treatment group were higher than Mdl group, but the difference was not statistically significant(P>0.05).except Ser group, the immobility time of the other groups were decreased compared with Mdl group.2. Results of E2 level:Compared with Mdl group. The level of serum E2 in S-M group and EB group was significantly increased (P<0.01).3.Results of RT-PCR:Compared with Ctl group, the mRNA expression of ERα、 ERβ, BDNF, TrkB were down regulated in the hippocampus of Mdl rats(P<0.01). Compared with Mdl group, the level of ERa. ERβ, BDNF mRNA increased in S-H、 S-M、S-L、EB group,TrkB mRNA Increased in S-M group, S-L group, EB group (P<0.01, P<0.05).4. Results of IHC:The ERα、ERβ protein expression in Mdl rats were lower than Ctl group(P<0.01).Compared with Mdl group.The protein expression of ERa in the hippocampus of each group increased in different degrees(P<0.0 1,P<0.05). Compared with Mdl group,The protein expression of ERβ in the hippocampus of S-H、S-M、S-L、EB group increased in different degrees (P<0. 01, P<0.05),among them, the most obvious increase was the S-M group.5.Results of WB:The BDNF、TrkB protein expression in hippocampus of Mdl group were lower than Ctl group(P<0.05, P<0.01). the protein expression of BDNF in S-H group,S-M group, EB group wre higher than the Mdl group (P<0.05, P<0.01). The protein expression of TrkB in S-H group,S-M group,S-L group, EB group wre higher than the Mdl group (P<0.01).Conclusion:1.Hormone stimulated pregnancy may provide a valuable animal model of PPD after hormone withdrawal. The model simulated the incidence of PPD in the etiology and behavior.This model can provide a good research platform for further exploring the pathogenesis of PPD.2.Shen Gui Ren mixture has the function of tonify deficiency and remove blood stasis. It has significant moderating effect on the Depression Behavior of PPD model rats.3. The change of serum E2 levels, hippocampal ERa, ERβ, BDNF, TrkB mRNA and protein of PPD model rat may be the important basis of the development of postpartum depression.4.The treatment of postpartum depression by Shengguiren mixture may improve the expression of estrogen and its receptor in hippocampus. Its anti-depression effect may be through the induction of BDNF, TrkB and other gene expression.5.The effect of middle dose of Shengguiren mixture is better than high dose and low dose, so it can be considered that middle dose (3.6g/kg.d) as the best effective dose.