| 1 background:Depression is a chronic mental illness,with significant and persistent low mood as the main clinical features,mainly for depression,depression and low self-esteem,mental retardation,and even be pessimistic and worldweary,suicidal,and has the characteristics of high morbidity,high disease burden,Gao Fufa rate,high disability rate,high rate of Dutch act etc..The syndrome of liver depression and spleen deficiency is a common clinical syndrome in Chinese medicine.Modern medicine advocates "disease dif-ferentiation and treatment",which advocates recognizing the disease from the whole process of disease.However,traditional medicine advocates "syndrome differentiation and treatment"to judge the location and nature of the disease from the current manifestation of the disease,so as to seize the main contradiction.Due to the different emphasis of "differentiation of dis-ease or treatment" or "syndrome differentiation and treatment",this paper explores the de-pression and spleen asthenia syndrome of depression from the perspective of "combination of disease and disease" from the perspective of "macro" and "micro".Astrocyte(Ast)is the most widely distributed type of cell in the central nervous system.With the further study of Ast function,it has been found to play a more and more important role in the occurrence and development of depressive disease.At present,the research of Ast has become a hot spot in the study of the pathophysiological mechanism of depression in so-cial stress and the target of preventing and controlling depression.In addition,some studies have found in the number of young early-onset depression in patients with cerebral Ast and its specific cytoskeletal protein GFAP expression was significantly decreased,while the ex-pression level of GFAP was significantly increased with the increasing of the age,the normal control group in older patients with brain age matched comparison this change was not ob-served significant.The results seem to suggest that the pathogenesis of GFAP in depression is different in both early and late depressive disorder.Glutamate is the most important excitatory transmitter in mammalian brain.It is con-verted from glucose to alpha ketosglutaric acid through the production of three carboxylic acid,and can also be hydrolyzed by glutamine.GlU is not evenly distributed in the central nervous system,with the highest content in the cerebral CORTex,hippocampus,cerebellum and striatum,second in the brain stem and hypothalamus,and more in the spinal cord.Gam-ma aminobutyric acid is the most important inhibitory neurotransmitter in the brain.It is produced by L-carboxyl decarboxylation,which is rich in brain.GABA neurons are distrib-uted in various layers of cerebral CORTex and cerebellar CORTex,striatum,hippocampus and so on.These brain regions are closely related to emotional and mental activity.The mag-netic resonance imaging(MRI)study and postmortem examination of patients with monoph-asic depression and manic depression found that glutamate energy loops were abnormal.Xiaoyao Powder is a classic Chinese medicine used in the treatment of liver qi stagnation and spleen deficiency syndrome.It has been widely used in the clinical and experimental study of liver depression and spleen deficiency syndrome.Animal experimental study group in our laboratory has been engaged in the depression of liver stagnation and spleen deficiency syndrome,has established a rat model of chronic restraint and CUMS,found that the classical Ming Xiaoyaosan has a more significant role in the regulation of the Xiaoyaosan can interfere with a variety of neural pathways,multi targets,multi role orientation.However,the current mechanism is still unclear whether GFAP and NMDA are regulated by regulation.2 objective:This paper through the animal model of rat CUMS is unpredictable stress depression copy of liver stagnation and spleen deficiency,first through the confirmation and change of CORT behavior model is successful,then the change of real-time fluorescent quan-titative PCR and WB experiments to further explore the GFAP and NMDA receptor,and ob-serve the changes of Xiaoyao Powder the "macro" behavior,including eating and weight changes and micro index.3 method:This topic consists of four parts,First,through combing the literature on de-pression in ancient literature,we initially comb the etiology,pathogenesis,pathogenesis and rationale and prescriptions of depression from the theoretical level,in order to provide theo-retical basis and support for experimental research.Two:on the basis of the experience and understanding of the subject group,the rat model of the liver depression and spleen deficien-cy of the rats was replicated by the method of CUMS,that is,unpredictable stress.The gen-eral physical condition of the rats was observed,such as body weight,food intake,open field and the behavioral changes in the sugar water experiment,and the changes of CORTi-costerone(CORT)in blood.And the traditional Chinese medicine Ming Fang Xiaoyao Pow-der and Western medicine for the treatment of depression were treated with fluoxetine in CUMS rats.Three:experiment two used real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)to detect the expression of GFAP and NMDA2B in the hippocampus of rats.Meanwhile,we observed the moderating effect of Xiaoyao San and fluoxetine.It is a certain exploration from the gene.Four:in experiment three,the expression of GFAP and NMDA2B in hippocampus of rat hippocampus was detected by immunoblotting(WB).In addition,the intervention of Xiaoyao Powder and fluoxetine was further summarized and discussed.4 results:Experiment one:the first successfully established CUMS rat model of chronic unpre-dictable stress,rats gradually appear under stress reduced food intake and weight gain de-creased,the hair becomes yellow,dry stool,reduce body fatigue activity does not adjust physical appearance changes of the state body syndrome rats;behavior study show:the total distance and the central movement of the rats in the open field test,the residence time de-creased gradually,reduce the rate of sugar sucrose preference test;micro index detection:el-evated serum CORT levels.While the Xiaoyao Powder group and the Qi Xi Ting group were significantly improved compared with the model group.Experiment two:on the basis of experiment after six weeks of chronic unpredictable stress rats were harvested after the detection of micro level indicator genes,the experimental results show that the rats in the normal group,model group and drug group rat GFAP and NMDA2B mRNA have a more significant difference 22,there were statistically significant differences.Experiment three:on the basis of Experiment 1,protein level was detected after sam-pling.In the detection of GFAP,there was a significant decrease in the model group com-pared with the normal group and the drug delivery group.In NMD2B,the model group was significantly higher than the normal group and the drug group,and the results were statisti-cally different.5 experimental conclusionsIn theory,the ancient books of depression were carefully discussed and arranged,and the syndrome of liver depression and spleen deficiency was the most common syndrome,but the deficiency of liver Yang Qi and kidney yang deficiency could not be ignored in the middle and severe stages of depression.The behavior and macro signs in the first experiment showed that the experiment suc-cessfully replicated the rat model of CUMS chronic unpredictable mild stress liver depression and spleen deficiency syndrome.The rat model was similar to emotional depression(depres-sive like behavior),depression and depression,abdominal distention,low stool(slow weight growth and lower food intake).While the traditional Chinese medicine Xiaoyao Powder and the classical western medicine for the treatment of depression,the reversal of the indexes af-ter the intervention of fluoxetine hydrochloride was also proved to be the successful estab-lishment of the model.In the detection of real time fluorescence quantitative PCR in experiment two,the expres-sion of NR2B in the model group was significantly reduced.The comparison between the model group and the Xiaoyao Powder and the fluoxetine group showed significant difference(P<0.01).The results showed that the expression of NR2B was down regulated by the chronic stress of CUMS in the rats with depression liver depression and spleen deficiency,but it was in the Chinese medicine compound Xiaoyao Powder and the western medicine fluoxetine.The expression of NR2B was improved to some extent.The GFAP in the gene model group was significantly different from the traditional Chinese medicine group and the western med-icine group(P<0.05),indicating that Xiaoyao Powder and fluoxetine could improve the damage of astrocytes and GFAP caused by chronic stress to a certain extent.The results of experiment three Westernblot showed that the protein expression of NR2B in the hippocampus of the model group decreased as compared with the normal group.After the treatment of Xiaoyao Powder and fluoxetine,the protein content of NR2B increased in varying degrees(P<0.05),and the difference was statistically significant.It indicates that Xiaoyao San and fluoxetine can increase the expression of NR2B induced by chronic stress.The expression of GFAP was significantly reduced in the model group.After the treatment of Xiaoyao Powder and fluoxetine,a significant increase was found,and the groups were statis-tically significant(P<0.05)compared with the model group(P<0.05),indicating that Xiaoyao Powder and fluoxetine could improve the expression of GFAP in the hippocampus of rats.These experiments confirm the hypothesis that the CUMS chronic stress causes the in-crease of the end product CORT of the HPA axis in the model group,which leads to the mod-erate activation of the Glu system and the increase in the adaptability of the body.However,as the stress continues to continue,the hippocampus deity is damaged and the astrocyte GFAP is reduced.In the end,it leads to the occurrence of depression.Xiaoyao Powder and fluoxetine are commonly used in the treatment of depression and spleen deficiency syndrome.Through experiments,we can see that Xiaoyao Powder and fluoxetine can significantly in-crease the expression of GFAP and NR2B genes and proteins in the "microcosmic" level.Under the intervention of Xiaoyao Powder and fluoxetine,the rats also appear in the aspects of macroscopic characterization and behavior.A certain improvement. |
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