| Part 1 Observation on the therapeutic effect of Huazhuo Jiedu Shugan Formula on patients with mild to moderate depression.Objective: To Study the effects of Huazhuo Jiedu Shugan Formula(HJSF)on the patients with mild to moderate depression.Methods:A total of 48 patients(24 in the treatment group and 24 in the control group)were diagnosed as mild to moderate depression with TCM turbidity toxin accumulation syndrome.The therapeutic effect was evaluated by HAMD score and TCM symptom scoreResults: 1.HAMD score showed that the scores of traditional Chinese medicine group and western medicine group were significantly reduced after treatment,and there was no difference between the two groups.2.TCM symptom score showed that the scores of TCM group and western medicine group were significantly reduced after treatment,and there was no difference between the two groups.The total effective rate of TCM group was 90.5%,which was better than that of western medicine group.Conclusion: 1.Huazhuo Jiedu Shugan Formula has an antidepressant effect.The effect on depression is similar to that of western medicine fluoxetine.2.The effect of Huazhuo Jiedu Shugan Formula on the improvement of TCM symptoms is similar to that of fluoxetine,but there are fewer adverse reactions in TCM group.Part 2 Observation of the effect of Huazhuo Jiedu Shugan Formula on thebehavior and the ultrastructure of neurons of chronic unpredictable mild stress(CUMS)miceObjective: To investigate the effect of Huazhuo Jiedu Shugan Formula on the behavior of CUMS mice,the ultrastructure of neurons,and to verify the antidepressant effect of this recipe through animal experiments.Methods:48 male ICR mice were randomly divided into normal group,model group,western medicine group and Chinese medicine high,medium and low dose group.CUMS model was established to observe the degree of anxiety and depression in mice through sugar water preference test,open field test,forced swimming test and elevated cross maze test,and observe the ultrastructure of neurons in the hippocampal CA3 region of mice by light microscopy and electron microscopy.Results: 1.In general,mice in the model group were observed to have dark hair color,timid and easily frightened,slow reaction,reduced activity,low spirit,reduced food intake,and even stress defecation.2.The sugar water preference index in the model group was significantly decreased compared with that in the normal group.3.The open field test scores of mice in the model group were significantly decreased,and both the traditional Chinese medicine group and the western medicine group were improved.4.In the model group,the total immobility of forced swimming was significantly prolonged,and the incubation period of immobility was shortened.5.Compared with the normal group,the visits and residence time of the open arm were significantly reduced in the model group,while the visits and residence time of the closed arm were significantly increased.6.light microscope: the nerve cells in the model group were disordered in arrangement,with nuclear pyknosis,more apoptotic bodies,and more glial cells and inflammatory cells.In TCM group,nerve cells were well arranged,with slight nuclear pyknosis,a small number of apoptotic bodies,and a small number of glial cells and inflammatory cells.7.Electron microscope showed that the number of neurons reduction,fuzzy structure,incomplete cell membrane,mitochondria swelling and vacuolar degeneration,chromatinaggregation,synapse numbers reduction in hippocampal CA3 area of the model group rats,which were alleviated with HJSF treatment.Conclusion: 1.The depression model of mice can be established successfully by CUMS and separation,and the model is stable and reliable.2..Huazhuo Jiedu Shugan Formula can improve the depression degree of CUMS mice.The group with high dose of traditional Chinese medicine has the best effect,which is equivalent to fluoxetine.3.It was confirmed from the aspect of morphology that CUMS can cause the damage of neurons in the hippocampal CA3 area in mice.4.Huazhuo Jiedu Shugan Formula can protect and reverse the neuron damage in the hippocampal CA3 area of mice,so as to restore the hippocampal structure,and provide theoretical basis for the discussion of the pathogenesis of depression from the perspective of morphology.Part 3 Observation of the effect of Huazhuo Jiedu Shugan Formula on the neuroplasticity of CUMS(chronic unpredictability mild stress)mice via Rac1-LIMK1-cofilin pathway.Objective: To investigate the expression of BDNF and receptor in CUMS mice and the possible mechanism of anti-depression effect of Huazhuo Jiedu Shugan FormulaMethods: 48 male ICR mice were randomly divided into normal group,model group,western medicine group and Chinese medicine high,medium and low dose group.The levels of BDNF,TrkB,SYP,PSD-95,Rac1,LIMK1,p-cofilin were detected by immunohistochemistry,western blot and real-time quantitative PCR.Results: 1.The expression of BDNF and TrkB in hippocampal CA3 were decreased in the model group,and Western Blot results further confirmed the immunohistochemical results.Q-PCR results showed that BDNF mRNA level in the model group was significantly lower than that in the normal group,and the expressions were increased in the high and medium dose groups of traditional Chinese medicine and the western medicine group.2.In the modelgroup,the expression of SYP and PSD-95 in hippocampus CA3 decreased.Western Blot results further confirmed the immunohistochemical results.The Q-PCR results showed that the SYP and PSD-95 mRNA levels in the model group were significantly lower than those in the normal group,which was consistent with the above results.The expression levels in the high and medium dose groups of traditional Chinese medicine and the western medicine group were increased.3.The expression levels of Rac1,LIMK1 and p-cofilin in the hippocampus of the model group decreased,while the expression levels increased in the high and medium dose groups of traditional Chinese medicine and the western medicine group.Conclusion: 1.Chronic unpredicted mild stress can successfully induce depression-like behavior in mice,accompanied by changes in hippocampal BDNF and receptors,synaptic related proteins and Rac1-LIMK1-cofilin pathway proteins.2.The mechanism of depression-like behavior in mice may be related to the protein changes in the Rac1-LIMK1-cofilin pathway mediated by BDNF-TrkB.3.Huazhuo Jiedu Shugan Formula has neuroprotective effects by affecting the protein changes in the Rac1-LIMK1-cofilin pathway mediated by BDNF-TrkB,thus producing an antidepressant effect. |
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