Regulatory Study On The Imbalance Of Intestinal Flora In UC Rat Model By Mongolian Medicine

Ulcerative colitis (ulcerative colitis, UC) is a primary, chronic rectum and colon mucosa disease. Ulcerative colitis (UC) is one of the most intractable diseases at present. Recently, the incidence of UC increases year by year. Now human’s health and their quality of life are more and more seriously influenced by UC. Western medicine mainly use the way of anti-inflammatory and immune suppression to cure the UC. But the side-effect is big, the response rate is low and the recurrence rate is high. Traditional usually use compound decoction and/or enema treatment to cure the disease and the treatment effect is good, but the patient adherence is poor. Chinese medicine is not easy to used for a long time on the treatment of UC. The Mongolian medicine for the treatment of chronic bowel disease has a long history and good curative effect. With long-term clinical observation, the research group found that the Garidi powder has good effect, low toxicity, and hardly relapse after cure. According to the theory of traditional Mongolian medicine and its application background, our study use modern separation and analysis techniques to extract and separate out effective parts from Papaver nudicaule L and Artemisia frigida Willd. And we also study the Garidi powder’s role of correcting intestinal flora imbalance in UC model rat.Method:1. Optimizing the extraction method of effective parts from Garidi powder.(1) The determination of factors and levels:Select different ethanol concen trations, the volume of solvent, reflux reaction time, reflux times. Each fa ctor set three levels.(2) Determine the contents of total alkaloids and flavonoids:use the Ultraviolet-visible spectrophotometry to determinine the flavonoids content, use non-aqueous titration/potentiometric titration method to determine the total alkaloids content in the extracting solutions and use LC-MSn to determine the alkaloids types.2. The research on the mechanism of Garidi powder’s function in correcting the UC model rat intestinal flora imbalance.The molecular target was intestinal bacteria’s 16SrRNA. The template was the total DNA separated from the rat colon contents. The intestinal flora fingerprint was drawn by capillary electrophoresis combined with terminal restriction fragment length polymorphism. We discussed the relationship between the intestinal flora variation and pathogenesis of ulcerative colitis. At the same time the experiment revealed Garidi powder’s function in correcting the UC model rat intestinal flora imbalance.Result:1. According to the actual conditions of production and cost, the optimum extraction conditions of Garidi Powder was 70% ethanol,22 times the volume of medicine plants,5 hours on extracting and 1 time for extracting.2. According to diversity index, HaeⅢ enzyme digestion samples’microbial diversity is higher than the HhaⅠ and MspⅠ enzyme digestion samples’. Therefore, we used the results of HaeⅢ enzyme digesting for the following analysis.3. The result of one-way analysis of variance showed that the intestinal fl ora which have statistic significance are Halomonas sp. wp29, uncultured Acidobacteria bacterium XB3A1, uncultured actinobacterium NLS2.9, uncult ured bacterium(T-RF:102bp), uncultured bacterium JH-WH31 and unculture d bacterium W23.Conclusion:1. According to Orthogonal array test, the experiment used visible spectrophotometry, non-aqueous titration and LC-MSn to optimize the extraction method for effective parts from Garidi Powder. According to the actual conditions of production and cost, the optimum extraction conditions of Garidi Powder was 70% ethanol,22 times the volume of medicine plants,5 hours on extracting and 1 time for extracting.2. The UC model rat’s intestinal flora imbalance are related with the foll owing two kinds of bacteria:Halomonas sp. wp29 and uncultured bacteriu m W23. The Garidi powder can maintain the UC model rat’s intestinal flo ra balance by adjusting the colonization of Halomonas sp. wp29, unculture d Acidobacteria bacterium XB3A1, uncultured actinobacterium NLS2.9, unc ultured bacterium(T-RF:102bp), uncultured bacterium JH-WH31 and uncultu red bacterium W23.