發(fā)布時間：2018-05-25 13:45

  本文選題：嗜酸乳桿菌上清 + 長雙歧桿菌上清��； 參考：《天津醫(yī)科大學》2017年碩士論文

【摘要】：目的:腸易激綜合征(Irritable Bowel Syndrome,IBS)是一種以腹痛、腹脹、腹部不適為特征癥狀,常伴有排便頻率和(或)糞便性狀改變的臨床常見消化道功能性疾病。近年研究表明,5-羥色胺(5-hydroxytryp-tamine,5-HT)及其轉(zhuǎn)運體(Serotonin Transporter,SERT)的代謝紊亂可能導致內(nèi)臟感覺異常、腸道運動紊亂,在IBS的發(fā)病中有重要作用。目前研究表明SERT表達的高低除受SERT基因多態(tài)性的影響外,還受腸道微生態(tài)的影響。大量臨床試驗證明益生菌可以改善IBS患者腹痛、腹脹等癥狀,對于治療腸易激綜合癥療效顯著。乳酸桿菌和雙歧桿菌是IBS中最常研究的益生菌,其中嗜酸乳桿菌及長雙歧桿菌已經(jīng)作為益生菌制劑應用于臨床。然而益生菌制劑改善IBS癥狀的具體作用機制尚不明確。我們之前的研究表明鼠李糖乳桿菌上清可以上調(diào)腸道上皮細胞和小鼠結(jié)腸組織SERT的表達,然而目前尚無研究證實嗜酸乳桿菌和長雙歧桿菌上清是否也能夠調(diào)節(jié)SERT的表達。本研究的目的是觀察嗜酸乳桿菌和長雙歧桿菌上清對腸道上皮細胞SERT mRNA及SERT蛋白的影響,從而為研究益生菌治療IBS作用機制提供新的理論依據(jù)。方法:本實驗分別用1:100、1:50、1:20 3種不同稀釋濃度的嗜酸乳桿菌(ATCC4356)上清和長雙歧桿菌(ATCC 15707)上清刺激HT-29和Caco-2細胞12小時和24小時,然后應用RT-PCR檢測細胞的SERT mRNA的表達水平變化,應用Western-Blot檢測細胞的SERT蛋白的表達水平變化。統(tǒng)計學分析應用SPSS 17.0統(tǒng)計軟件:用均數(shù)±標準差(±s)來表示計量資料。應用單因素方差分析來比較3組以上計量資料的組間差異,用LSD方法檢驗方差齊性數(shù)據(jù),用Dunnett方法檢驗方差不齊數(shù)據(jù)。以P0.05表示差異有統(tǒng)計學意義。結(jié)果:1:100,1:50和1:20稀釋的嗜酸乳桿菌上清刺激HT-29細胞12小時和24小時,刺激組SERT mRNA表達水平分別是對照組的1.80,2.24,2.28,和2.04,2.30,2.80倍(P0.05,P0.05,P0.05,和P0.05,P0.05,P0.05);1:100,1:50和1:20稀釋的嗜酸乳桿菌上清刺激Caco-2細胞12小時和24小時,刺激組SERT mRNA表達水平分別是對照組的1.20,1.22,1.61,和1.16,1.50,2.13倍(P0.05,P0.05,P0.05,和P0.05,P0.05,P0.05);1:100,1:50和1:20稀釋的長雙歧桿菌上清刺激HT-29細胞12小時和24小時,刺激組SERT mRNA表達水平分別是對照組的0.96,1.90,2.50,和2.58,2.19,3.21倍(P0.05,P0.05,P0.05,和P0.05,P0.05,P0.05);1:100,1:50和1:20稀釋的長雙歧桿菌上清刺激Caco-2細胞12小時和24小時,刺激組SERT mRNA表達水平分別是對照組的1.25,2.06,1.83和1.84,2.23,2.13倍(P0.05,P0.05,P0.05,和P0.05,P0.05,P0.05)。各濃度嗜酸乳桿菌與長雙歧桿菌上清均上調(diào)HT-29細胞和Caco-2細胞SERT蛋白的表達,上調(diào)水平與相應的mRNA上調(diào)水平大致相同。結(jié)論:(1)嗜酸乳桿菌及長雙歧桿菌上清可以上調(diào)腸道上皮細胞SERTmRNA及SERT蛋白的表達;(2)嗜酸乳桿菌及長雙歧桿菌上清上調(diào)SERTm RNA和SERT蛋白的表達具有時間依賴性和濃度依賴性。
[Abstract]:Objective: Irritable Bowel Syndrome syndrome (IBS) is a common clinical gastrointestinal functional disease characterized by abdominal pain, abdominal distension and abdominal discomfort, often accompanied by defecation frequency and / or fecal changes. Recent studies have shown that the metabolic disorder of serotonin transporter serotonin (serotonin) and serotonin transporter (serotonin) may lead to visceral sensory disorder and intestinal motility disorder, which may play an important role in the pathogenesis of IBS. Current studies have shown that the expression of SERT is affected not only by the polymorphism of SERT gene, but also by intestinal microecology. A large number of clinical trials have proved that probiotics can improve abdominal pain, abdominal distension and other symptoms in patients with IBS, and it is effective in the treatment of irritable bowel syndrome. Lactobacillus and Bifidobacterium are the most commonly studied probiotics in IBS. Lactobacillus acidophilus and Bifidobacterium long have been used as probiotics in clinic. However, the mechanism of probiotics in improving IBS symptoms is unclear. Our previous studies have shown that the supernatants of Lactobacillus rhamnoides can up-regulate the expression of SERT in intestinal epithelial cells and colonic tissues of mice. However, there is no study on whether the supernatants of Lactobacillus acidophilus and Bifidobacterium can also regulate the expression of SERT. The purpose of this study was to observe the effects of supernatants of Lactobacillus acidophilus and Bifidobacterium longidus on SERT mRNA and SERT protein in intestinal epithelial cells, and to provide a new theoretical basis for studying the mechanism of probiotics in treating IBS. Methods: in this experiment, HT-29 and Caco-2 cells were stimulated with 1: 100 and 1: 50: 20 supernatants of 1: 100, 1: 50, 1: 20 with different diluted concentrations of Lactobacillus acidophilus (ATCC4356) and Bifidobacterium longidus (ATCC15707) for 12 and 24 hours, respectively. The expression of SERT mRNA was detected by RT-PCR. The expression of SERT protein was detected by Western-Blot. Statistical analysis using SPSS 17.0 statistical software: mean 鹵standard deviation (鹵s) to represent the measurement data. Univariate analysis of variance was used to compare the differences among three groups of measurement data. LSD method was used to test the homogeneity data of variance and Dunnett method was used to test the uneven data. The difference was statistically significant with P0.05. Results HT-29 cells were stimulated by the supernatant of Lactobacillus acidophilus diluted at 1: 1: 100 and 1:20 for 12 hours and 24 hours, respectively. The SERT mRNA expression levels in the stimulated group were 1.80 / 2.242.28 and 2.042.302.80 times as compared with those in the control group, respectively, and the Caco-2 cells were stimulated by the supernatants of P0.05P0.05P0.05P0.05P0.05P0.05: 50 and 1:20 diluted by Lactobacillus acidophilus for 12 hours and 24 hours, respectively, in which the supernatant of Lactobacillus acidophilus diluted at 1: 1: 50 and 1:20 stimulated Caco-2 cells for 12 hours and 24 hours, respectively. The expression levels of SERT mRNA in the stimulated group were 1.20 ~ 1.22 / 1.61, 1.16 ~ 1.50 ~ 1.50 ~ 2.13 times, P _ (0.05) P _ (0.05) P _ (0.05) and P _ (0.05) P _ (0.05) P ~ (0.05) respectively, and the supernatant of 1: 100: 1: 50 and 1:20 diluted bifidobacterium longus stimulated HT-29 cells for 12 and 24 hours, respectively. The expression levels of SERT mRNA in the stimulated group were 0.96 ~ 1.90 ~ 2.50, 2.58 ~ 2.19 and 3.21 times of the control group, respectively, and the levels of SERT mRNA expression in the stimulated group were as follows: P0.05, P0.05, P0.05, and P0.05P0.05, respectively, and the SERT mRNA expression levels in the stimulated group were 12 hours and 24 hours after the supernatant of P0.05: 100: 50 and 1:20 diluted by Bifidobacterium longii, respectively. The SERT mRNA expression levels in the stimulated group were 1.252.061.83 and 1.82.232.13 times of those in the control group, respectively. The levels of SERT mRNA expression in the stimulated group were 1.252.06.83 and 1.82.232.13 times that of the control group, respectively, and the expression levels of SERT mRNA in the stimulated group were 1.252.06.83 and 1.82.232.13 times as much as that in the control group. The supernatants of Lactobacillus acidophilus and Bifidobacterium longidus upregulated the expression of SERT protein in HT-29 and Caco-2 cells, and the up-regulation level was approximately the same as that of mRNA. Conclusion the supernatants of Lactobacillus acidophilus and Bifidobacterium longifera can up-regulate the expression of SERTmRNA and SERT protein in intestinal epithelial cells. The supernatants of Lactobacillus acidophilus and Bifidobacterium can up-regulate the expression of SERTm RNA and SERT proteins in a time-and concentration-dependent manner.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R574.4

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