Study On Effects And Mechanisms Of Wendan Decoction In Rats With Schizophrenia

objective:Wendan decoction (WDD in abbreviation) is widely used in clinic, especially in depressive and manic psychosis which is similar to schizophrenia in western medicine.To treat the schizophrenia by TCM, most physicians would start with phlegm and attach an importance to regulating qi to resolve phlegm. WDD is a classic formula for treating the psychiatric diseases induced by the invisible phlegm. Our experiments are to systematically investigate the effects and mechanisms of WDD through the 2 pathogenesis of schizophrenia respectively----hypofunction of Glutamic acid (Glu in abbreviation)and hyperfunction of Dapamine (DA in abbreviation),to investigate effects of WDD on learning-memory, synapse long-term potentiation (LTP in abbreviation) in model rats with schizophrenia and effects of WDD on changes of Glu, DA and metabolite in cerebral cortex, hippocampus and corpus striatum, to observe influence of WDD on expression of receptor D2 of DA in model-rats’corpus striatum, Glu in hippocampus, subunit NR1 and NR2B of N-methyl-D-aspartate receptor and protective effects of pathological injuries,further, to look into the effects of WDD on dynamic changes of free radical,nitric oxide (NO), T cell subclusters,interleukin-2(IL-2 in abbreviation) and to observe effects of WDD on immune-regulating and effects of WDD against oxidation injury, All the experiments mentioned above try to preliminarily investigate net-regulating effects of WDD on Glutamic acid----Gama aminobutyric acid----DA neuron and to explore the relationship among oxygen-derived free radicals, NO,and immunization and their role in psychiatric diseases, whishing the experiments can provide new suggestions and ideas for therapeutic methods and pathogenesis of schizophrenia and afford reliable, systematical experimental foundations for clinical application.Methods:The therapeutic effects and mechanisms of WDD were systematically studied by utilizing model rats with different schizophrenia types induced by MK801-receptor antagonist of NMDA and APO--receptor agonist of DA.1. The effects of WDD on model rats’learning memory function were observed by Morris water maze--examining methods of behavior science, the electronic physiological observation on the functional plasticity of PP-DG synapse and its manifestations in model rats with schizophrenia were carried out, which may expound the mechanism of WDD to improve model rats’learning memory.2.By the methods of High performance liquid chromatograph--fluorescence detection(HPLC-FD) and electrical chemistry detection (HPLC-ECD), the contents of Glu, GABA, Aspartic acid (ASP),DA and its metabolites were determined to investigate the regulating effects of WDD on Glu and DA system in model rats.3.By the changes of immunohistochemistry and neuropathological morphology we observed influence of WDD on expression of receptor D2 of DA in model-rats’NA,Glu in hippocampus,receptor subunit NR1 and NR2B of N-methyl-D-aspartate and protective effects of pathological injuries on model rats’hippocampus.4.Through the experiments of DA receptor binding, We observed the effects of WDD on the affinity of D2 receptors and the number of expression and explored the effects and mechanisms of WDD against schizophrenia by the receptor level.5.To investigate effects of WDD on immune-regulating and effects of WDD against oxidation injury in model rats with schizophrenia by determination of activities of SOD,MDA, CAT and NO, by quantitative determination of TC subcluster (CD3,CD4 and CD8) through flow cytometer (FCM) and by determination of serum IL-2 through ELISA.Results:1.The results of behavioral science examination shows that in the Morris water maze, compared with the model group, the latent period of rats in WDD group was significantly shorten and the frequency of bestraddling-dais was remarkably increased.In the experiment of transmission efficacy of synapse, WDD could improve the transmission efficacy of dentate gyrus of hippocampus,raise the PS value of dentate gyrus of model rats induced by HFS,shorten peak of the latent period, further, increase the maintenance ability LTP, indicating WDD may improve learning-memory through enhancing the plasticity of transmission efficacy of synapse,increasing the contents and functions of Glu, preventing the neuron cell in hippocampus from denaturing or delay the denaturalization to improve the ability of learning memory. 2.Compared with the normal group and WDD group,The contents of Glu and ASP of prefrontal cortex and hippocampus in model rats with schizophrenia induced by MK801--receptor antagonist of NMA were significantly lower, however, the contents of GABA is remarkable higher, and the contents of DA, DOPAC,HVA of corpus striatum in model rats remarkable higher those of normal group and WDD group, whereas, the contents of Glu and ASP in WDD group were higher than those of other gropus, and the DA and its metabolites were lower those of other groups, indicating that there are changes in Glu and DA system which can interact each other in model rats with schizophrenia and that WDD can regulate the dysfunctions of Glu and DA system in model rats’ brain.3.The results of morphological observation show that there are significantly differences in expression of Glu, subunit NR1 and NR2B of NMDA receptor in all groups and there also are significantly differences in cell structure and cell denaturalization of CA1 area in hippocampus.WDD can improve disorder of cell rearrangement,relieve the process of nuclei dissolution, postpone the denaturalization of hippocampus cell,indicating WDD has certain protective effects of pathological injury in model rats’hippocampus.4.The classic behavior such as smelling, biting,licking and so on arose in all the model rats induced by APO,However, the rats in normal group did not have such behavior, compared with model group, such behavior in WDD group inhibited 20 minutes latter, indicating WDD has the effect of against classic behavior induced by APO.D2 receptor competitive binding experiments shows that WDD can decrease number of D2 receptor, raise the affinity with D2 receptor, indicating WDD can prevent schizophrenia from attacking through hindering D2 receptor.5.Compared with Normal group and WDD group, the plasma contents of SOD and CAT in model group were lower,the contents MDA were higher, while in WDD group the contents of SOD and CAT increased to certain extent indicating WDD can eliminate free radical in model rats, relieve the peroxidation injury. In addition, The NO activity of model group is higher than those of normal group and WDD group, indicating that WDD prevents NO participating in neuro-toxicity of ASP through decreasing activity of NO. 6.The determinations of FCM and ELISA shows there are difference in contents of CD3,CD4 and IL-2 among all groups, the contents of CD3+,CD4+,CD4+/CD8+ in model group are significantly lower those of normal group and WDD group, however, there are no significantly difference among all the groups, the contents of CD3+,CD4+,CD4+/CD8+ and IL-2 increase,indicating that WDD can regulate the contents of TC subcluster and IL-2,and suggesting WDD can reinforce integral immunity of model rats with schizophrenia through contributing clone proliferation of particular T cells and increasing number of T cells.Conclusion:To sum up, WDD has better therapeutic effects on model rats with schizophrenia. Its mechanism may correlate with regulating dysfunctions and imbalance of Glu and DA in brain, enhancing the plasticity of hippocampus synapse, protecting hippocampus from injury, relieving the harm resulting from oxygen-derived free radicals, improving the immunoregulation.Our research affords WDD systematically experimental foundation for further exploitation.