| Treatment with syndrome differentiation as one of the characteristics of Traditional Chinese Medicine indicates the integrated , dynamic change and individual conception of TCM. With the development of biomedicine,the research of genomics and proteomics comes to the most important orientation.The functional disorder of genes and proteins cause the disease, the expression and regulation of genes and proteins are similar to the basic theory of TCM. So the research of syndrome focus on the technology of genomics and proteomics.Liver depression is one of common syndromes in clinical practice.As a complex symdrome,liver depression relates to the complex diseases of other zangfu organs,including nerve, endocrine, circulation, digestion, immunity, feeling and motion etc are involved. However, only part of known gene expression products (receptor, enzyme, cytokine and peptides etc) change is disclosed so far in some aspects, essence of liver stagnation syndrome is far away from uncovering.ObjectIn order to explore the essence of liver depression deeply,we chose the gene GSK-3βwhich maybe relate to liver depression we found in researches before,to find the sequential changes of it in the hole experiment:Methods1. Establish the Rat Model of Liver-Depression Syndrome by Chronic Unpredictable Mild Stress. There were 3 experiments in the whole research, the time last of the models are 1 week.2 weeks and 3 weeks.We used 32 male SD rats in every experiment,group the rats with a randomized and regional grouping design to 4 groups: the normal control group, the group of stress model , the group of stress model administered with Xiaoyaosan,the group of stress model administered with amitriptyline.2. The following day after the stress, we test the wastage of 1% saccharose solution of 1 hour and calculate the increasement of the weight.3.After the experiment, the rats were put to death and the serum was collected to mesure the corticosterone concentrations with ELISA Kit.4.After the experiment,the rats were put to death,apart the hippocamps on ice,then extracted the total RNA with Trizol,Amplificat the gene GSK-3βwith Reverse Transcription Nested PCR, to analyze the relative value of GSK-3βwith Phoretix 1D software.Results1. In the whole experiments ,compare with the normal control group ,the stress model rats were down in spirit ,the growth of weight were slower,the fur lost of gloss generally,less active,most time attached the wall.2. In the 1 week and 2 weeks models wastage of 1% saccharose solution in 1 hour had not significant differences;after 3 week's CUMS,the wastage of 1% saccharose solution in 1 hour between the normal rats and stress rats were significant different.It seems as that the copy of the liver depression model are successful. The differences of the weight increase are not regulate.3. The difference of the corticosterone concentrations in 1 week and 3 weeks model are not significant;in the 2 weeks model the corticosterone concentrations of stress rats are significant lower than other 3 groups.4. We can't find exactly variation in the relative value of GSK-3β,the result has not achieved our anticipation.ConclusionsBy evaluating the 1% saccharose solution in 1 hour,we can come to the conclusion that the copy of the liver depression model is successful basically,but the relative value of GSK-3βdon't have significant differences between model rats and normal rats.There are some reasons for the result:first of all,there are some mistakes during the experiment;secondly,there are some differences between animals and experiment situations;thirdly,there are more complex mechanisms of the gene expression in the liver depression, GSK-3βis not the most important gene relate to liver depression.All the above need deeper researchment in following days. |
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