本文選題：慢傳輸型便秘 + 腸道微生物群落結構��； 參考：《昆明理工大學》2017年碩士論文

【摘要】：慢傳輸型便秘(slow transit constipation,STC)是臨床常見的慢性頑固型便秘,占便秘發(fā)病率的25%。其臨床表現(xiàn)頑固,綜合保守治療效果不佳。由于臨床濫用瀉劑,導致便秘的進行性加重,最終導致藥物失效只能實施手術治療。由于該疾病的發(fā)病機制至今尚不明確,給臨床防治帶來極大困難。本研究使用分子生物學方法結合高通量測序分析慢傳輸型便秘及益生菌制劑緩解慢傳輸型便秘過程中糞便樣品中的微生物群落結構,并對構建的慢傳輸型便秘小鼠生理指標進行分析,研究益生菌混合發(fā)酵酸奶對慢傳輸型便秘的治療效果,以期為臨床診斷及治療提供理論依據(jù)。小鼠及便秘患者實驗結果表明,自主配方三菌酸奶可明顯緩解慢傳輸型便秘癥狀。通過對小鼠生理指標的檢測,可以看出三菌酸奶組小鼠的小腸推進率、首次排黑便時間均優(yōu)于其他組,且三菌酸奶可明顯逆轉(zhuǎn)小鼠的便秘癥狀。而通過小鼠糞便樣品中短鏈脂肪酸(short-chain fatty acids,SCFAs)的檢測可以看出三菌酸奶有明顯的降低能量供應的作用。從OTU數(shù)目、ACE和Chaol指數(shù)來看,隨著喂食酸奶天數(shù)的增加,小鼠糞便樣品及慢傳輸型便秘患者糞便樣品中的細菌多樣性顯著降低,而小鼠在喂食便秘藥物洛哌丁胺后,兩菌酸奶組小鼠OTU數(shù)目與健康對照組增加且無顯著差別,而三菌酸奶組與便秘模型組降低。慢傳輸型便秘患者糞便樣品中的細菌多樣性顯著降低,這說明便秘導致腸道中的有害微生物增加,而可能導致其他一些微生物的生長受到抑制甚至死亡,喂食益生菌混合發(fā)酵酸奶后,有益微生物在腸道內(nèi)大量定植,抑制了有害微生物的生長,從而使糞便樣品中細菌的多樣性呈現(xiàn)下降的趨勢。在小鼠實驗中,比較不同組所有小鼠的細菌群落結構顯示,在門的分類水平上,四個組的小鼠糞便樣品中主要的細菌為Bacteroidetes、Firmicutes和Proteobacteria。飲用兩菌酸奶組小鼠糞便樣品中Firmicutes與Proteobacteria的量明顯低于其他組,而Bacteroidetes所占序列比例更高。隨著實驗天數(shù)的增加,各組小鼠糞便樣品中的微生物群落發(fā)生著明顯的改變。對不同組別的小鼠按處理時間進行微生物群落分析可以看出,小鼠糞便樣品中Candidatusfamily ratAN060301C、Rikenellacae隨著喂食小鼠酸奶天數(shù)的增加而增加,而Lachnospiraceae、Desulfovibrionaceae 與 Carnobacteriaceae 是減少的,其之所以在糞便樣品中增加或減少是由于其在腸道內(nèi)的定植發(fā)生改變。UPGMA(unweighted pair-group method with arithmetic means)聚類和主成分分析(principal components analysis,PCA)都表明,不管何種方法,所得到的細菌群落聚類結果大致相同,喂食益生菌制劑前小鼠糞便樣品中的細菌群落結構分布不均勻,無顯著相關性,而喂食益生菌制劑后小鼠糞便樣品中細菌群落結構較為相似,且三菌酸奶組比兩菌酸奶組更為相似。比較所有便秘患者飲用益生菌混合發(fā)酵酸奶前后糞便樣品中細菌群落結構,可以看出,在門的分類水平上,飲用益生菌混合發(fā)酵酸奶前糞便樣品中的細菌多樣性高于飲用后,而其中有近一半的序列屬于Bacteroidetes和Firmicutes,Proteobacteria和Actinobacteria序列所占比例較少。在屬的分類水平上,飲用益生菌混合發(fā)酵酸奶前糞便樣品中的細菌多樣性高于飲用后,但兩個樣品中優(yōu)勢屬所占的序列百分比并無明顯差異,均為Bacreroides和Faecalibacterium。通過對每個患者飲用益生菌混合發(fā)酵酸奶前后糞便樣品中細菌的主要屬進行的分析和比對可以看出,絕大多數(shù)患者飲用益生菌混合發(fā)酵酸奶前后糞便樣品中的優(yōu)勢菌均為 Bacteroides,大多數(shù)是 Faecalibacterium。研究慢傳輸型便秘患者飲用益生菌混合發(fā)酵酸奶前后糞便樣品微生物群落結構組成和演替規(guī)律,結合小鼠便秘模型試驗,尋找引起便秘及益生菌混合發(fā)酵酸奶緩解便秘過程中的關鍵微生物,為便秘的臨床治療提供有力的新的治療手段。
[Abstract]:Slow transit constipation (STC) is a common chronic and stubborn constipation in clinic. The clinical manifestation of 25%. in the incidence of constipation is obstinate, and the effect of comprehensive conservative treatment is not good. Because of the clinical abuse of laxatives, the progressive constipation is aggravated and the drug failure can only be performed by operation. The system is still not clear to date and brings great difficulties to clinical prevention and treatment. This study uses molecular biological methods and high throughput sequencing to analyze slow transmission constipation and probiotics to alleviate the microbial community structure in the process of slow transmission constipation, and analyze the physiological indexes of the slow transmission type constipation mice. The therapeutic effect of probiotic mixed fermented yogurt on slow transit constipation was provided in order to provide theoretical basis for clinical diagnosis and treatment. The experimental results of mice and constipation patients showed that the spontaneous formulation of three bacteria yogurt could obviously relieve the symptoms of slow transmission constipation. By testing the physiological indexes of mice, the small intestine of the three bacteria yogurt mice could be seen. The rate of intake was better than the other groups for the first time, and the three bacteria yogurt could obviously reverse the symptoms of constipation in mice. Through the detection of short-chain fatty acids, SCFAs in the fecal samples of mice, it was found that three bacteria yogurt had a significant reduction in energy supply. From the number of OTU, ACE and Chaol index, the feed was fed with feeding. The increase in the number of yogurt days, the bacterial diversity in the fecal samples of the mice and the slow transit constipation patients decreased significantly, while in the mice fed with the constipation drug luperidine, the number of OTU in the two bacteria yogurt mice increased and no significant difference was found, while the three bacteria yogurt group and the constipation model group decreased. The bacterial diversity in the fecal samples is significantly reduced, which indicates that constipation leads to the increase of harmful microbes in the intestinal tract, which may lead to the inhibition or even death of some other microorganisms. After feeding the probiotics to the mixed fermented yogurt, the beneficial microbes colonize the intestinal tract and inhibit the growth of harmful microorganisms, thus causing feces. In the mice experiment, the bacterial community structure of all the mice in the different groups showed that the main bacteria in the four groups of mice fecal samples were Bacteroidetes, Firmicutes and Proteobacteria. in the fecal samples of the two bacteria yogurt mice, and Firmicutes and Pro in the fecal samples of the mice. The amount of teobacteria was significantly lower than that of the other groups, and the proportion of Bacteroidetes was higher. With the increase of the experiment days, the microbial community in the fecal samples of each group changed obviously. The microorganism community analysis of the mice in different groups could be seen, and the Candidatusfamily ratA in the mice feces samples. N060301C, Rikenellacae increased with the increase in the number of yoghurt days in feeding mice, while Lachnospiraceae, Desulfovibrionaceae and Carnobacteriaceae were reduced, and the reason for the increase or decrease in fecal samples was due to the change of.UPGMA (unweighted pair-group method with arithmetic means) clustering in the intestinal tract. Principal components analysis (PCA) and the principal component analysis (analysis) showed that the results of the bacterial community clustering were almost the same regardless of the method. The bacterial community structure in the fecal samples of the pre feeding probiotic preparation was not uniform, and the bacterial community structure in the fecal samples of the mice after feeding probiotics was fed. The three bacteria yogurt group was more similar than the two bacteria yogurt group. Compared with the bacteria community structure in the fecal samples before and after the mixed fermented yogurt of all constipation patients, it can be seen that the diversity of the bacteria in the fecal samples before the mixed fermentation of the yogurt was higher than that in the faecal samples before the mixed fermentation of the yogurt. Half of the sequences belong to Bacteroidetes and Firmicutes, and the proportion of Proteobacteria and Actinobacteria sequences is less. In the classification level of the genus, the bacterial diversity in the fecal samples of the mixed fermented yoghurt before drinking is higher than that after drinking, but there is no significant difference in the percentage of the dominant genera in the two samples, all of which are Bacreroid By analyzing and comparing the main genera of the bacteria in the fecal samples before and after the mixed fermented yogurt of each patient, ES and Faecalibacterium. can see that the dominant bacteria in the fecal samples of the overwhelming majority of patients who drank the probiotic mixed fermented yogurt were Bacteroides, most of which were the slow transmission of Faecalibacterium.. The microbial community structure and succession law of the fecal sample before and after the mixed fermentation of probiotics in the constipation of the constipation patients, combined with the mouse constipation model test, to find the key microbes in the process of constipation causing constipation and probiotic mixed fermented yogurt to relieve constipation, and provide a powerful new therapeutic means for the clinical treatment of constipation.

【學位授予單位】：昆明理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R574.62

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