The Study Of Xiaoyao San On Improving Hippocampal Neuroplasticity In Depressed Mice

Depression is a common clinical disease that seriously endangers the physical and mental health of human beings.In recent years,the incidence rate has been continuous increasing,which seriously threatens people's physical and mental health and has a negative impact on people's daily life and work.However,there are obvious adverse reactions and side effects due to antidepressants,then the research on this field has good clinical significance and application prospects.Traditional Chinese medicine treatment of emotional diseases,the classic Fang Xiaoyao San treatment of depression,the curative effect is exact,and its related mechanism deserves further research and discussion.ObjectiveThis experiment aims to confirm that Shugan Jieyu Recipe Xiaoyao Powder can regulate the expression of its molecular chaperone protein(FKBP5),enhance the activity of glucocorticoid(GR)and up-regulate the brain source by acting on chronic stress-induced hippocampal neuronal cells.The expression of neurotrophic factor(BDNF)can improve the neuroplasticity of hippocampus,and attempts to reveal the molecular and molecular mechanisms of the effect of liver-relieving and relieving stagnation by regulating neuroplasticity and exerting anti-stress effects.MethodsExperiment 1(Animal experiment)A chronic stress model of FKBP5 knockout mice was established by chronic unpredictable stress(CUMS).Gene knockout mice were randomly divided into control group,chronic stress group,Xiaoyao San intervention group,low,medium and high dose group and GR receptor blocker intervention group.Behavioral indicators were measured in each group of mice,serum glucocorticoid levels were measured by ELISA,and changes in hippocampal synaptic structure and neuronal regeneration were detected by stereological methods.HE staining and MAP-2 fluorescence immunohistochemistry were performed.Learn to observe.The expressions of GR,BDNF and FKBP4/FKBP51 mRNA in hippocampus were detected by RT-PCR.The total protein,cytoplasm and GR protein expression in GR,BDNF and FKBP4/FKBP51 in hippocampus were detected by Western Blot.Immunoprecipitation was detected by immunoprecipitation.FKBP4/FKBP5 combined with GR.The mechanism of Xiaoyao Powder on hippocampal neuroplasticity in rats with chronic stress injury was studied from the experimental level in vivo.Experiment 2(cell experiment)used glucocorticoids(GCs)to establish a model of stress cell injury.The primary cultured neurons for 7-8 days were divided into low-dose control group of corticosterone+Xiaoyao San,low-dose stress model of corticosterone+Xiaoyao Powder,corticosterone+Xiaoyao San+FKBP5 low expression hippocampal neurons,corticosterone+Xiaoyao San+FKBP5 overexpression,corticosterone+GR receptor agonist,corticosterone+GR receptor blocker intervention group.Normal light microscopy and inverted phase contrast microscopy were used to observe the morphology of neurons in 1 day and 7 days,CCK8 to detect cell proliferation activity,and EDU to detect cell proliferation.Western blot was used to detect primary cultured neurons GR,p-GR,BDNF and Changes in protein and mRNA expression of FKBP5.From the in vitro experimental level,the mechanism of Xiaoyao Powder on hippocampal neuroplasticity induced by glucocorticoids was discussed.ResultsExperiment 1(animal experiment)1.The activity distance of the mice in the model group was reduced,and the rest time was prolonged,which was significantly different from the blank control group(P<0.05).Compared with the model group,the activity distance and activity of each Xiaoyao Powder group were higher,and the rest time was lower than that of the control group.The statistical difference was obvious.(P<0.05).2.The sucrose preference of the model group decreased,which was statistically significant compared with the blank control group.(P<0.05).The sucrose preference of the low-medium-high group and the mifepristone group in Chinese medicine increased significantly(P<0.05).3.The results of Western-blot showed that pGR,GR in the model control group compared with the blank control.,BDNF average optical density value decreased.Compared with the model control group,Xiaoyao was significantly up-regulated pGR,GR,and BDNF average optical density values(P<0.05).4.Quantitative PCR results showed that the expression levels of BDNF mRNA were significantly decreased in the stress model group and each drug-ad?? istered group compared with the blank control group.Experiment 2(Cell experiments)1.When FKBP5 is overexpressed,it can inhibit cell activity,and when it interferes,cell activity is enhanced.2.When FKBP5 is overexpressed,the increase of GR into the nucleus increases,and when it interferes,the nucleus decreases.Moreover,when the expression of FKBP5 is interfered,the expression of BDNF in the cells is increased,which also indicates that FKBP5 overexpression has a certain protective effect on damaged cells.ConclusionShugan Jieyu Recipe Xiaoyao Powder can regulate the expression ofFKBP5 in hippocampal neurons of rats with chronic stress injury,enhance the activity of GR into the nucleus,up-regulate the expression of GR target gene BDNF,and improve the neuroplasticity of hippocampus,thereby exerting anti-stress injury effect.Overexpression of FKBP5 can affect the nuclear activity of GR,leading to increased expression of BDNF,thereby exerting protective effects on damaged cells.