The Anti-anxiety And Mechanism Study Of Baixiangdan Capsule On The Model Rats With Anxiety

Objective: To study the the anti-anxiety effect of Baixiangdan capsules, differentmethods were selected to establish a rat model with anxiety. This research will study thebasic pharmacodynamic of anti-anxiety of the Baixiangdan capsule based on selecting asuitable modeling method, the paper. Using appropriate modeling methods of anxiety ratsand BXD best effect dose, by comparing the model group and treatment group serumnoradrenaline (NA), brain-derived neurotrophic factor (BDNF) levels and brainα1A-adrenergic receptor (α1A-AR), BDNF protein expression changes, reveal BXDanxiolytic effects at the molecular level possible drug targets and mechanism.Method: Establish rat model by uncertain empty bottle stimulation method and footshock method respectively, evaluate each rat group’s behavior by open field test andelevated plus maze test, and observe the anti-anxiety effects of Baixiangdan capsule. Studythe drug effect gradient of low (100mg·kg-1·d-1), medium (200mg·kg-1·d-1) and high (400mg·kg-1·d-1) doses on Baixiangdan capsule’s anti-anxiety effect in appropriate rat models ofanxiety and select diazepam (1mg·kg-1·d-1) as positive control. Detecting the serum NA andBDNF levels of each rat group by ELISA method, detecting the brain region α1A-AR andBDNF protein expression of each rat group by Western Blot method. Experimental datawill be analyzed by using the software SPSS17.0and the figures for the statistical analysisresults will be made by the software Graph Pad Prism5.0.Results:1. Uncertainty empty bottle stimulation method and foot shock method are chosen tobuild anxiety rat model, and the experimental results of the open field experiments and theelevated plus-maze(EPM) experiments shows that the total path of the rats in model groupis longer than the rats in blank control group in open field experiments(P<0.01), and thepercentage of entries into open arms(OE%) and of time spent on open arms(OT%) decrease(P<0.01). Both of uncertainty empty bottle stimulation method and foot shockmethod are the effective anxiety rat modeling method, and the preliminary determinationresults of pharmacodynamics shows that the Baixiangdan (BXD) capsule have anxiolyticeffect. By contrasting the two models, the anxiety symptom caused by the foot shockmethod can preferably simulate the clinical manifestation of anxiety neurosis and is somature and reliable that it can be used to do the experiment on the drug screening on amass scale. As a result, this study choose the foot shock method to build anxiety rat modelfor the subsequent experiments on the study of the pharmacodynamics and its mechanism.2. Produce the rat model of anxiety by footshock method and evaluate the dose-effectrelationship for anti-anxiety effects of the Baixiangdan capsule. By contracting to the ratsof blank control group, the total path of rats in the model group increase(P<0.01) in openfield test, the percentage of entries into open arms(OE%) and of time spent on openarms(OT%) decrease(P<0.01) in EPM(Elevated Plus-Maze) test. By contracting to themodel group, the total path of the BXDII group and DZP group decreasesignificantly(P<0.01), and both the OE%and OT%increase in EPM. However the totalpath, OE%and OT%of BXDI group and BXDIII increase and decrease seperately, buttheres’s no statistical significance. The results show that BXD has a marked anxiolyticeffect in middle dosage (200mg·kg-1·d-1), but the anti-anxiety effect of BXD do notincrease or decrease by increasing the dosage of BXD.3. BXD capsule can affect the NA and BDNF level in the serum and encephalic regionof the rats in the anxiety model which is built by the foot shock method. The ELISA resultsshow that, contracting to the blank control group, the NA level in the serum of the modelgroup increases(P<0.01) and the BNDF level decreases(P<0.01). After intervention ofdrugs, the NA level in serum of the DZP group and BXDII group is lower than the modelgroup(P<0.05), the BNDF level is higher than model group(P<0.05). The western blotresults show that, the expression level of BNDF protein in the left hippocampal, the righthippocampal of model group is lower than blank control group(P<0.05or P<0.01), theexpression level of BNDF protein in the left hippocampal, the right hippocampal of BXDIIgroup and DZP group is higher than the model group(P<0.05or P<0.01). The expressionlevel of the BNDF protein in the frontal lobe and the hypothalamus of model group ishigher than the blank control group(P<0.01), the he expression level of the BNDF proteinin the frontal lobe and the hypothalamus of BXDII group and DZP group is lower than themodel group(P<0.01).The expression level of the α1A-AR protein in each encephalic region(the left hippocampal, the right hippocampal, the frontal lobe and the hypothalamus)of the model group is higher than the blank control group(P<0.01), and the expression levelof the α1A-AR protein of the BXDII group and DZP group is lower than modelgroup(P<0.05or P<0.01).Conclusions:1. Both the uncertainty empty bottle stimulation method and foot shock method couldbuild anxiety rat model, chinese medicine Baixiangdan could improve anxiety-likebehavior and has a good anti-anxiety effect. However, the foot shock method is an idealmethod to build the anxiety rat modeling method for its good reproducibility, mature,reliability and applicability in the experiment on the drug screening on a mass scale. Thepresent research observes the anti-anxiety effect of BXD from unconditioned response andconditioned response and the reliability of pharmacodynamic evaluation increases.2. The evaluation results of the dose-effect relationship for anti-anxiety effects of theBaixiangdan capsule show that, the total path in open field experiments of model groupincreases than the blank control group(P<0.01), and the percentage of entries into openarms(OE%) and of time spent on open arms(OT%) decrease(P<0.01) in EPM tes. The totalpath,OE%and OT%of the medication administration team is higher or lower than themodel group seperately and BXDII group (8times than the clinical dose) has a significantdifference than others(P<0.01), but the anti-anxiety effect do not increase or decrease byincreasing the dosage.3. By studing the mechanism of BXD capsule, the resulth shows that BXD capsulecan resist anxiety neurosis by reducing the level of the NA and improving the level ofBNDF in serum while improving the expression level of BNDF in the hippocampal,reducing the the expression level of BNDF in the frontal lobe and the hypothalamus orreducing the expression level of α1A-AR protein in central, BDNF and α1A-AR may bethe effect target of Baixiangdan capsule. The findings could provide the clues of the effecttarget and mechanism for the Chinese herbal compound.