The Study Of Macrophage-related Factors Associated With Behaviors In Depressed Mice And The Regulation Of Acupunture And TCM Treatment

Depression is a kind of syndrome with significant and lasting depressed mood as the main clinical features, and a common mental disease jeopardizing people's health. Involved in the pathogenesis of nervous, endocrine, immune, and many other changes, its exact pathogenesis is still unclear. In addition, we seldom discussed the depression-related changes of cytokines during the development of depression in previous studies, and rarely reported the changes of related cytokines during the treatment of depression, while also knowing little about the immunological mechanism in the process of forming and development of depression. This project aims to study the changes of macrophage-related cytokines in the development of depression, and treat it with Chinese medicine to investigate the immunological mechanism.The research includes:1.The changes of related macrophage cytokine in chronic unpredictable mild stres (CUMS) modelCUMS was employed to induce depression in mice. Mice were given a certain intensity of random stimulation in the continuing 21 days:fasting, tilt feeding, shaking horizontally, cultivating wetly, reversing day and night, lighting overnight, clamping tail, cold stimulation, thermal stimulation, noise stimulation, etc. Open-field test and sucrose preference test were operated on model mice after modeling,at the same time the measure of mice weight and food intake were needed,as a index to evaluate the results of the modeling. The results showed that, with the model of chronic unpredictable mild depression developing, the weight of model mice (14 d,21 d) decreased significantly than the control mice; the food intake of model mice also showed a declining trend, compared with the control group. Significant differences were observed after 21 days since beginning, so 21 days can form a model of depression.The peritoneal macrophages were Isolated from modeling mice under sterile conditions, in order to use ELISA method and MTT method to measure the changes of contents of NO, TNF-a, IL-lβ, IL-6 and other cytokine in the supernatant fluid of peritoneal macrophages after they were cultured and stimulated with LPS. The results showed that NO, secreted by macrophage, changed little in the first 3d and 7d, but decreased significantly in the 2Id. The ability of TNF-a's secretion had the trend to improve meanwhile the ability of IL-iβ's secretion in the middle and late time (7d,2Id) increased significantly, and ability of IL-6's secretion improved obviously in the later time (21 d) in modeling.2. The effects of acupuncture combined with Fluoxetine (i.g.) on improving behaviour and on macrophages in CUMS depression.Mice were given treatment of acupuncture combined with Fluoxetine (i.g.) to chronic unpredictable depression modeling mice, and observe the changes of their behaviour in different time, and determine the relevant peritoneal macrophage cytokine content. The results showed that, after 7 days'treatment, the fluoxetine group and acupuncture combined group mice could both improve the horizontal movements, sugar preferences and such behaviour related to depressing. After 14 days'treatment, the modeling animals had a certain self-recovery capability, performing as improvements on the opening behavior. However, sucrose preference still remained on low levels, and the treatment did not show any effects of improving. After 14 days' intervention treatment, both groups have shown significant improvements in mice sucrose preference. After 7 days and 14 days treatment, the ability of secreting NO by macrophages significantly increased. After 7 days'treatment, acupuncture combined group's secretion of macrophage cytokines TNF-a, IL-lβ, IL-6 decreased significantly. After 14 days'treatment, the cytokine TNF-a, IL-6 decreased significantly, as well IL-lp showed decline trend.3. The effects of Ganmai Dazao Tang on improving CUMS and on macrophages.Use Ganmai Dazao Tang to treat the mice, observe its impact on mice behaviour, determine mice peritoneal macrophages cytokine contents, and discuss the immune mechanisms of Chinese medicine treatment on depression. We found that, after seven days'treatment, Ganmai Dazao Tang showed significant effect on the sugar preference rate, fluid consumption, and weight of mice, and after fourteen days'treatment, the sugar preference rate has been significantly improved. The mice secretion of NO of Ganmai Dazao Tang group in both 7d and 14d also showed a significant improvement, similar as fluoxetine. The secretion of IL-6 by macrophages decreased significantly after seven days'treatment with Ganmai Dazao Tang, while TNF-a and IL-lβsecretion also showed the same trend, but decreased little. After 14 days'treatment with Ganmai Dazao Tang, the ability of secreting TNF-a and IL-6 by macrophages had significant declined.We can conclude from the above results that, in the process of CUMS depression, the ability of modeling mice secreting cytokines by peritoneal macrophages have changed, performing as the increase of the ability of secreting TNF-a, IL-6, IL-lp, and the decrease of ability of secreting NO, after the modeling is completed. Indeed, It indicates inflammation and immune response activation in the formation of depression.Traditional Chinese medicine (acupuncture combined with Fluoxetine (i.g.) and Ganmai Dazao Tang) can significantly improve the chronic unpredictable modeling mice behaviour of depression, develop the mice's ability to respond to the happy events. At the same time, it can increase the secretion of NO by mice peritoneal macrophages, while reduce the ability of secreting TNF-a and IL-6, and affect the capacity of the secretion levels of IL-lβ(with no significant difference). The regulation effect of drug therapy suggest further that macrophage related cytokines have an important role in the pathogenesis of depression.