| Depression is a common mental disorder that has complex clinical manifestations. Because of there are many predisposing factors of depression, the pathogenesis of depression has been unclearly explained. Epidemiological survey has found that depression patients are not only displaying mental disorder, but also accompanying with irritable bowel syndrome, functional dyspepsia and other gastrointestinal dysfunction. However, the present theoretical hypothesis could not explain this phenomenon. To clarify the pathogenesis of depression, it is an urgent need to find new ideas and methods. As an important environmental factor, intestinal microbiota plays an important role in gastrointestinal function, and it has became a new hot spot of disease, so it is necessary to discuss the relationship between intestinal microbiota and depression.In this thesis, we, with some emphasis, discuss three problems around intestine microbiota in depression situation as following:①To establish ERIC-PCR method to study intestinal microflora in depressed situation by optimizing sample preparation, genomic DNA extraction and ERIC-PCR amplification, compare the polymorphism of intestinal flora in different parts of intestine tract, and analyze whether the structure of intestinal flora was changed in chronic unpredictable mild stress (CUMS) model rats after the intervention of Xiao yaosan.②To establish PCR-DGGE method of intestinal microflora in depressed situation by optimizing 16S rDNA PCR amplification, compare the structure of intestinal flora in CUMS model after the intervention of drugs, and identify different strains.③Using paraffin embedding, RT-PCR and GC-MS techniques, analyze alteration of inflammatory response and chemical constituent in intestinal tissue in CUMS model rats after the intervention of drugs.The results showed that:①Animal model of depression is successfully induced by CUMS, and Xiao yaosan and Fluoxetine Hydrochloride have significant antidepressant effects. In addition, unnormal performance of caecum was also observed in Model rats, such as increase of cecal weight and cecal index, and the intervention of Xiao yaosan could alleviate this effect.②Caecum could be used as a new material owing to the most significant polymorphism of cecal microbiota. Compared with Healthy Control (HC) group, ERIC-PCR fingerprinting was changed significantly in Model group, but recoveried in treatment group.③Compared with HC group, PCR-DGGE fingerprinting was changed significantly in Model rats, but varied in treatment group. Sequence analysis results showed that the bacteria strains were Lachnospiraceae bacterium, Lactobacillus animali, Burkholderiales bacterium and Lactobacillus reuteri.④Cecal tissue had no significant abnormalities in HC group. It had changed in Model rats, but no obvious inflammatory cell infiltration. The treatment group had some improvement.⑤Compared with HC group, gene expression related to inflammatory response were up-regulated in Model rats. After the intervention of Xiao yaosan and Fluoxetine Hydrochloride, down regulated gene expression were.⑥Compared with HC group, the chemical composition of cecal tissue in Model rats was changed, but the treatment group showed a different characteristics.In summary, we established ERIC-PCR and PCR-DGGE methods for studying intestinal microflora in animal model of depression, and found that CUMS could significantly change the diversity of intestinal microbiota and morphology, histochemisty and gene expression related to inflammatory response in caecum. And Xiao yaosan and Fluoxetine Hydrochloride could not only adjust intestinal microflora, but also Xiao yaosan had roles in prevention of inflammatory response. Besides, caecum may play a role of preventing organ in depression. Changes of these indicators except histochemisty could be used as supplementary information to evaluate occurrence of depression. |
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