Study On The Antidepressant Effect And Molecular Mechanism Of Mongolian Medicine

[Objective]l.No depression of the classical literature of Mongolian medicine records, but according tothe incidence of external factors and typical clinical manifestations and symptoms,depression and heart "Heyi" and "Badagan" is closely related to the. Mongolian medicine believes that relationship between the heart and the mind and spirit is most closely, depression is the disease of heart. So the study of Mongolian medical theory, combined with the depression of the domestic and foreign latest research developments and trends, the choice has the improved efficacyof blood running, sedative traditional Mongolian medicine ZADI-5 pills, through study the effect on CUM rats hippocampus, monoamine neurotransmitters, neurotrophic factors andcytokines in serum the anti depression effect, clarify ZADI-5 pills and its possible mechanism.2.By means of modern technology, combined with the complexity of the pathogenesis of depression, depressive mechanism of anti multi way to study ZADI-5 pills, further confirm its antidepressant effect in the view of molecular biology, and provide the experimental basis for clinical application and research.3.Further explore the etiology and pathogenesis of depression from the perspective of Mongolian medicine, fully embodies the theory and method of diagnosis and treatment intraditional Mongolian medicine and pharmacy is unique, and lay the foundation for thestudy of mechanism of action of antidepressant drugs provide the clinical application and development of new ideas and ZADI-5 pills.[Methods]1.Grouping and administration in rats:The 60 SD rats were raised after 1 week according to body weight were randomly divided into 6 groups,10 rats in each group. They are normal group, model group, positive control group, ZADI-5 pills in high dose group, low. The normal group and each cage 5, free drinking and eating, do not accept any stimulation. The remaining group of single cage feeding, and accept the stimulation of 21d. At the same time of modeling, the rats were administered, the positive control group was given fluoxetine hydrochloride (3.6 mg/kg), ZADI-5 pills, in low, high dosage group (were given ZADI-5 pills 0.8g,0.4g,0.2g/ kg), normal group and model group were given equivalent physiological saline.2.The method of establishing model and evaluation model:By CUMS with the lonely raise replication model of depression. Stress at the end of a change in weight, rat Open-field activity detection experiment, 1% sucrose consumption test and Morris water maze learning and memory ability of detectionexperiments behavior detection and evaluation model.3.Preparation of samples and test index:After the end of behavior observation, anesthetized rats, abdominal aortic blood,3000r/min and centrifuged for 15 minutes, the supernatant was saved to be measured. The rats were decapitated rapidly after stripping the brain hippocampus, quick freezing with liquid nitrogen preservation to be measured.① The ELISA technique was used to detect thecontent of rat hippocampal 5-HT, DA, NE and monoamine neurotransmitter content and serum IL-1, IL-2, IL-6, IFN-a, IFN-r, TNF-a, ACTH, CORT cytokines.② The expression was detected by immunohistochemical method in rats hippocampus CAland CA3 region of BDNF protein.③ The expression uses the RT-PCR technology of BDNF in the rat hippocampus detectionmRNA.4.By using the SPSS 19.00 statistical software processing, all data with the average standard deviation x±s said, the selection of single factor analysis of variance, with P<0.05 asthe difference had statistical significance.[Results]1. Effect of ZADI-5 Pills on the behavior of CUM rats1.1 Effect on body weight changes in rats:Observation of stimulated rat weight changes slowly to the course of the experiment, in experiments seventh,14,21 days as compared to the control group, model group rats’body weight decreased significantly (P<0.05, P<0.01). During the fourteenth day experiment, compared with the model group rats’ body weightdose group rats ZADI-5 pills increased significantly (P<0.05). In the experiment twenty-first days, and the rats of model group was significantly highercompared to the positive control group and ZADI-5 pills in low dose group, increase of body weight (P<0.05, P<0.01). The test results show that, ZADI-5 pills and fluoxetine can improve the changes of body weight in CUM rats.1.2 Effects of rat 1% sucrose water drinking amount:Compared with the blank group, significantly reduced 1% sucrose water consumption of the rats in the model group (P<0.05). Compared with the model group,1% sucrose water consumption increased significantly isthe positive control group, ZADI-5 pills of rats in high dose group (P<0.05). The test results show that the improved CUMS rats,1% sucrose water consumption is ZADI-5 pills and fluoxetine.1.3 Effects on Open-Field activity in rats:Compared with the blank group, significantly reduced the level crossing case number andnumber is erected in the rats of model group (P< 0.01). Compared with the model group, the square crossing significantly increase is the positive control group, ZADI-5 pills high, medium dose group rats (P<0.05, P<0.01), the erection times significantly increased ZADI-5 pills of rats in high dose group (P<0.01). The test results show that, on the outside of the fresh environment curiosity ZADI-5 pills can improve CUM rats decreased1.4 Influence on learning and memory ability of rats:The 21 day stress over time, compared with the blank group, the escape latency of model group increased significantly (P< 0.01). Compared with the model group, the escape latency was shortened the fluoxetine and ZADI-5 pills high, low dose group rats (P<0.05,0.01), no significant differences in middle dose group, but there is the tendency to reduce certain. Compared with the blank group, model group, the average times across the platformreduced, but increased in varying degrees of rat model of average times across the platform colour of fluoxetine and ZADI-5 pills have no significant difference between the pill. The test results show that, ZADI-5 pills and fluoxetine can improve the learning and memory ability of CUM rats, has certain antidepressant effect.2. Effect of ZADI-5 pills on serum cytokines of CUM rats2.1 Effects on IL-1, IL-2, IL-6:Compared with normal group significantly increased the content of serum IL-1, IL-2, IL-6 of the rats in the model group (P<0.01). Compared with model group, positive control group and ZADI-5 pills high dose group rats serum IL-1 contents were significantly decreased (P<0,05); no significant differences in ZADI-5 pills low dose group, but there was a tendency of decrease.2.2 Effects on IFN-a, IFN-r, TNF-a:Compared with the normal group, the serum IFN-r, TNF-a content in rats of model group (P<0.01). Compared with the model group, significantly reduced the serum IFN-r, TNF-a content wasthe positive control group and ZADI-5 pills of rats in high dose group (P<0.05); there isa tendency to decrease but no significant difference is ZADI-5 pills, in low dose group.2.3 The influence on the CORT and ACTH:The model group rats serum ACTH and CORT content was obviously higher than normal group (P<0.01). Positive control group and ZADI-5 pills high, medium dose group rats serum ACTH andCORT content were significantly lower than those in the model group (P<0.05,0.01); no significant differences in ZADI-5 pills low dose group serum, but there is a tendency ofdecrease.3. Effects of Rat Hippocampal Monoamine transmitter:The rats in the model group were significantly lower than normal group in hippocampus 5-HT, NE, DA content (P<0.05, P< 0.01). The model group was significantly lower than that of the positive control grouphippocampus 5-HT, the content of NE (P<0.05), model group was lower than that in ZADI-5 pills high dose group, middle dose group of NE in the hippocampus and hippocampal 5-HT, DA content were increased significantly (P<0.05).4. The expression of BDNF protein in hippocampus of rats:Cause the change of BDNF expression in rat hippocampus CUMS, mainly in CA1, CA3 region, CA3 region which is more outstanding. The normal control group, positive cells in rat hippocampal CA1 and CA3 regions of densely arranged, zonal distribution.Hippocampal BDNF immune response in rats of model group decreased and the differencecompared with the normal control group statistically. Fluoxetine and ZADI-5 pills group,the expression of BDNF in hippocampus of rats was improved to different extent, which ZADI-5 pills, large dose group and fluoxetine group BDNF of the rat hippocampal CA3 region positive cells arranged in dense, zonal distribution, the number and percentage ofIOD positive cells increased significantly, compared with the model group, the differencethere were statistically significant (P<0.05, P<0.01).Dose group rats hippocampal CA13 area of BDNF positive cell number ZADI-5 pills inpercentage and IOD increased significantly, compared with the model group, the difference was statistically significant (P<0.05).5. nfluence on expression of BDNF mRNA in hippocampus of rats:Altered expression in hippocampus of rats caused by BDNF mRNA CUMS, and the normal control group was significantly decreased (P<0.05). Fluoxetine and ZADI-5 pills group of rat hippocampal BDNF mRNA expression increased in varying degrees (P<0.05, P<0.01), expression ZADI-5 pills low dose group of rat hippocampal BDNF mRNA one (P<0.05).[Conclusion]1. This study used the chronic unpredictable mild stress (CUMS), with the method of feeding alone replicate depression model, simulate the natural pathological process and the ability of learning and memory disorder, depression monoamine neurotransmitters decreasedactivation, immune cells, neurotrophic factors change due to characteristics of sub lack ofbehavior and pathology, with science and the reliability of the strong.2.Learn to ZADI-5 pills can improve the behavior of CUM rats, and can improve the Open- Field activity and the ability of learning and memory, has a role in the prevention and treatment of depression.3.ZADI-5 pills can improve CUM rat brain 5-HT, NE, DA and monoamine neurotransmitters,the effect may be related to the content of monoamine neurotransmitters.4.ZADI-5 pills can inhibit CUM rat serum IL-1, IL-2, IL-6, IFN-a, IFN-r, TNF-a, ACTH, CORT and other cytokine activation, its effect may exert antidepressant effects through immune suppression and regulation of HP A axis function.5.The expression of BDNF protein and mRNA ZADI-5 pills on hippocampal tissue in CUM rats with upregulation of nerve, has good protection of hippocampal neuron damage, repair and promote the regeneration. One of the important ways of this mechanism may be anti depressive mechanism of ZADI-5 pills.6.Traditional Mongolian medicine and Western medicine ZADI-5 pills Fluoxetine Hydrochloride Capsules on CUMS rats antidepressant effect comparison of results, ZADI-5 pills on the behavior and hippocampal neural endocrine, immune suppression, neuron nutrition protection effect was significant, suggesting that t ZADI-5 pills antidepressant usesimilar to fluoxetine, and as traditional and natural medicine, its toxic side effect, has thevalue of further research and development.