| Objectives:The anxiety model of rats was established by chronic restraint stress,and the effects of Jiaotai Pill on the general state,body weight and behavior of chronic restraint stress rats were observed.The anti-anxiety mechanism of Jiaotai Pill was discussed by detecting the expression levels of glucocorticoid receptor(Glucocorticoid receptor,GR),brain derived neurotrophic factor(Brain derived neurotrophic factor,BDNF),tyrosine kinase receptor B(Tropomyosin receptor kinase B,Trk B)and p-Trk B protein,as well as BDNF m RNA and Trk B m RNA in hippocampus of rats.Methods:Sixty male Wistar rats were selected and randomly divided into blank group,model group,diazepam group and low dose,medium dose and high dose groups of Jiaotai Pill according to body weight and open field test results,each group had 10 rats and were kept in cages.The blank group did not receive any stimulation,while the other groups used 14 days of chronic restraint stress to establish anxiety model of rats.Each dose group of Jiaotai Pill and diazepam group were given low dose Jiaotai Pill(0.75 g crude drug /kg),medium dose Jiaotai Pill(1.5 g crude drug /kg),high dose Jiaotai Pill(3 g crude drug /kg)and diazepam(diazepam,1 mg/kg)respectively before daily restraint.The blank group and model group were given the same amount of ultrapure water.The general state and body weight changes of rats were observed,and the open field test(the number of erections and total distance of movement)and light-dark transtition test(times of box penetration and open box residence time)were observed to reflect the anxiety state of rats.The expression levels of GR,BDNF,Trk B and p-Trk B proteins in rat hippocampus were determined by Western bloting.The expression levels of BDNF m RNA and Trk B m RNA in rat hippocampus were determined by RT-PCR.Results:1.Effect of Jiaotai Pill on General State and Weight of Chronic Restraint Stress Rats:After 14 days of chronic restraint stress,compared with the blank group,the rats in the model group had the symptoms of mental stress,easy manic anger,vertical and scattered fur,yellowing and darkening,preference for dormant corners,and irregular stool.The anxiety performance of each dose group of Jiaotai Pill and diazepam group was improved compared with the model group.Changes of body weight in rats:compared with the blank group,the weight of the rats in the model group decreased significantly(P<0.01).Compared with the model group,the weight of the rats in the diazepam group increased significantly(P<0.05),the weight of the rats in each dose group of Jiaotai Pill showed an increasing trend,but there was no statistical difference.2.Effect of Jiaotai Pill on Behavior of Chronic Restraint Stress Rats:In the open field test,after 14 days of chronic restraint stress,compared with the blank group,the rats in the model group have significantly reduced the number of erections and the total movement distance(P<0.01).Compared with the model group,the erect times of rats in the medium dose group of Jiaotai Pill and diazepam group increased significantly(P<0.05),the high dose and low dose groups of Jiaotai Pill showed an increasing trend,but there was no statistical difference.Compared with the model group,the total exercise distance of rats in the low,medium and high dose groups of Jiaotai Pill and diazepam group increased significantly(P<0.05 or P<0.01).In the light-dark transtition test,after 14 days of chronic restraint stress,compared with the blank group,the number of times the rats wear the box and the residence time in the bright box were significantly reduced(P<0.01).Compared with the model group,The number of times the rats in the low,medium and high dose groups of Jiaotai Pill and diazepam group wore boxes increased significantly(P<0.05 or P<0.01).Compared with the model group,the open box residence time of rats in the medium dose group of Jiaotai Pill and diazepam group were significantly increased(P<0.05).3.Effect of Jiaotai Pill on BDNF-Trk B Signal Pathway in Chronic Restraint Stress Rats:Effect of Jiaotai Pill on expression levels of GR,BDNF,Trk B and p-Trk B proteins in hippocampus of rats with chronic restraint stress:after 14 days of chronic restraint stress,the expression levels of GR,BDNF,Trk B and p-Trk B protein in hippocampus of rats in the model group were significantly lower than those in the blank group(P<0.01).Compared with the model group,the expression levels of BDNF and p-Trk B protein in hippocampus of rats in each dose group of Jiaotai Pill and diazepam group were significantly increased(P<0.05 or P<0.01).Compared with the model group,the expression level of GR protein in hippocampus of rats in the low dose group of Jiaotai Pill was significantly increased(P<0.05),theexpression level of GR protein in hippocampus of rats in the medium dose,high dose groups of Jiaotai Pill and diazepam group showed an increasing trend,but there was no statistical difference.Compared with the model group,the expression level of Trk B protein in hippocampus of rats in the low dose,medium dose groups of Jiaotai Pill and diazepam group was significantly increased(P<0.05 or P<0.01),while the expression level of Trk B protein in hippocampus of rats in the high dose group of Jiaotai Pill was increased,but there was no statistical difference.Effect of Jiaotai Pill on expression levels of BDNF m RNA and Trk B m RNA in hippocampus of chronic restraint stress rats:after 14 days of chronic restraint stress,compared with the blank group rats,the expression levels of BDNF m RNA and Trk B m RNA in hippocampus of model group rats decreased significantly(P<0.01).Compared with the model group,the expression level of BDNF m RNA and Trk B m RNA in hippocampus of rats in each dose group of Jiaotai Pill and diazepam group were significantly increased(P<0.05 or P<0.01).Conclusion: Jiaotai Pill can improve anxiety-like behavior of chronic restraint stress rats,which may be realized by improving the level of key factors in BDNF-Trk B signal pathway. |
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