Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用及对肠道菌群的影响

发布时间：2018-01-01 04:22

本文关键词：Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用及对肠道菌群的影响　出处：《南京中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的：探讨Faecalibacterium prausnitzii (.F. Prausnitzii,普拉梭菌)联合溃克灵防治炎症性肠病的疗效机制及对肠道菌群的影响。方法：将50只Sprague-Dawley大鼠随机分为5组：正常组、模型组、F.prausnitzii组、溃克灵组、F.prausnitzii联合溃克灵组,干预组于造模前各灌胃一周,采用2,4,6-三硝基苯磺酸(TNBS)法(80mg/KgTNBS+50%的乙醇)灌肠诱导结肠炎大鼠模型,观察大鼠的生存状态,于3天后全部处死,观察大鼠体重及结肠长度变化,并行结肠组织病理学评分；ELISA法检测各组大鼠血浆IL-10、IL-12、TNF-α、IFN-γ水平,计算IL-10/IL-12比值；免疫组化法检测肠粘膜IL-10、TNF-α的表达；实时荧光定量PCR法定量检测各组大鼠粪便中普拉梭菌、长双歧杆菌、嗜乳酸杆菌、大肠杆菌含量,并计算长双歧杆菌(B)与大肠杆菌(E)的比值(B/E值)。结果：1.造模后大鼠均出现腹泻症状,与正常组相比,模型组大鼠体质量明显下降,结肠明显缩短,结肠组织损伤明显(均P0.001)；与模型组相比,F.prausnitzii联合溃克灵组、溃克灵组Neurath积分降低(均P0.05), F.prausnitzii联合溃克灵降低了体质量的下降、结肠的缩短(均P0.05)。相对于模型组,各预防用药组大鼠血浆IL-10水平、IL-10/IL-12比值明显升高(均P0.05),其中以F.prausnitzii联合溃克灵组效果最为显著(P0.001,P0.003)；相对于模型组,各预防用药组血浆TNF-α水平无明显差异,F.prausnitzii联合溃克灵组大鼠肠粘膜IL-10表达升高(P0.05)、血浆IL-12下降(P0.05),各预防用药组肠粘膜TNF-α表达均下降(均P0.05)。F.prausnitzii联合溃克灵组、溃克灵组血浆IFN-y水平明显低于模型组(P0.001, P0.05)。与F.prausnitzii组、溃克灵组相比,F.prausnitzii联合溃克灵组大鼠结肠组织病理学评分降低, IL-10/IL-12比值及血浆IL-10水平、肠粘膜IL-10表达升高,血浆IL-12、IFN-γ、TNF-α水平及肠黏膜TNF-a表达下降,但差异无统计学意义。2.模型组大鼠粪便中普拉所菌、嗜乳酸杆菌含量较正常组均明显下降(P0.003,P0.05),模型组、溃克灵组、F.prausnitzii组大鼠粪便中大肠杆菌较正常组含量明显升高(P0.003或P0.01)。与模型组相比,F.prausnitzii组、F.prausnitzii联合溃克灵组大鼠粪便中普拉梭菌含量明显升高(P0.05,P0.003),溃克灵组普拉梭菌含量及各预防用药组长双岐杆菌、嗜乳酸杆菌的含量有升高,但均无统计学差异。与正常组相比,模型组、F.prausnitzii组长双歧杆菌(B)与大肠杆菌(E)的比值(B/E值)均下降(P0.001,P0.05)。F.prausnitzii联合溃克灵组与模型组相比,B/E值升高(P0.05),F.prausnitzii联合溃克灵组与F.prausnitzii组、溃克灵组相比,普拉梭菌、长双歧杆菌、嗜乳酸杆菌含量、B/E值均上升,大肠杆菌含量下降,但均无统计学意义。结论：益生菌F.prausnitzii联合中药溃克灵对结肠炎大鼠肠粘膜的预防保护作用较单药显著,可能与抑制炎症因子IL-12、TNF-α、IFN-γ分泌、促进抗炎因子IL-10表达的免疫调节作用相关；可以增加结肠炎大鼠肠道益生菌的含量、抑制致病菌的生长,提示F.prausnitzii联合溃克灵对纠正肠道菌群紊乱有一定的作用。
[Abstract]:Objective: To investigate the Faecalibacterium prausnitzii (.F. Prausnitzii, faecalibacterium prausnitzii) effect mechanism of combined effects of kuikeling treatment of inflammatory bowel disease and effect on intestinal microflora. Methods: 50 Sprague-Dawley rats were randomly divided into 5 groups: normal group, model group, F.prausnitzii group, kuikeling Decoction group, F.prausnitzii combined with kuikeling Decoction group, the intervention group before modeling each gavage for a week, using 2,4,6- three trinitrobenzene sulfonic acid (TNBS) method (80mg/KgTNBS+50% ethanol enema) induced rat colitis model rats, observe the survival of the state, all animals were killed in 3 days, to observe the changes of body weight and length of the colon of rats, parallel scores colon tissue pathology; to detect the plasma IL-10, ELISA IL-12, TNF- alpha, IFN- levels, IL-10/IL-12 ratio was calculated; immunohistochemistry was used to detect the expression of TNF- in intestinal mucosa IL-10; real-time fluorescence quantitative PCR to detect the legal amount of excrement At faecalibacterium prausnitzii, Bifidobacterium longum, Lactobacillus acidophilus, Escherichia coli, and Bifidobacterium longum (B) and Escherichia coli (E) ratio (B/E value). Results: 1. after the model rats showed symptoms of diarrhea, compared with the normal group, rats body weight decreased significantly. Colon colon tissue damage was significantly shortened (P0.001); compared with the model group, F.prausnitzii combined with kuikeling Decoction group, kuikeling Decoction group Neurath score decreased (P0.05), F.prausnitzii combined with kuikeling Decoction reduces body weight, shorten the colon (P0.05). Compared with the model group, preventive medication rats, plasma IL-10 levels, IL-10/IL-12 ratio increased significantly (P0.05), of which F.prausnitzii combined with kuikeling Decoction group was the most significant (P0.001, P0.003); compared with the model group, no significant difference between preventive groups, plasma TNF- levels, F.prausnitzii combined with kuikeling Decoction group The rat intestinal mucosa increased expression of IL-10 (P0.05), plasma IL-12 decreased (P0.05), the preventive medicine group of intestinal mucosa TNF- expression decreased (P0.05).F.prausnitzii combined with kuikeling Decoction group, IFN-y group of plasma Kui Keling was significantly lower than the model group (P0.001, P0.05). With the F.prausnitzii group, kuikeling decoction group F.prausnitzii, the joint effects of kuikeling rats colon tissue pathology score decreased, IL-10 ratio and plasma IL-10/IL-12 level, IL-10 expression in intestinal mucosa increased, plasma IL-12, IFN- gamma, TNF- alpha level and intestinal mucosal expression of TNF-a decreased, but the difference was not statistically significant.2. stool rats model in the Pula bacteria, Lactobacillus acidophilus compared with that in the normal group were significantly decreased (P0.003, P0.05), model group, kuikeling Decoction group, significantly higher than the normal group content of Escherichia coli in feces of rats in the F.prausnitzii group (P0.003 or P0.01). Compared with the model group, F.prausnitzii group, F.prausnitz II grams of faecalibacterium prausnitzii Kui Ling group content in feces of rats increased significantly (P0.05, P0.003), the Kui Keling group of faecalibacterium prausnitzii content and the preventive medication head of Bifidobacterium, Lactobacillus acidophilus with content increased, but there was no statistical difference. Compared with the normal group, model group, F.prausnitzii group of Bifidobacterium (B) Escherichia coli (E) and the ratio (B/E) decreased (P0.001, P0.05).F.prausnitzii combined with kuikeling Decoction group compared with the model group, the increase of B/E value (P0.05), kuikeling decoction combined with F.prausnitzii group and F.prausnitzii group, kuikeling Decoction group, faecalibacterium prausnitzii, Bifidobacterium longum, Lactobacillus acidophilus content the value of B/E, increased, decreased the content of Escherichia coli, but had no statistical significance. Conclusion: F.prausnitzii probiotics combined with traditional Chinese medicine collapse prevention protection spirit grams of the intestinal mucosa in rats with colitis significantly than single drug, may be related to the inhibition of inflammatory cytokines IL-12, TNF- alpha, gamma IFN- Secretion can promote the immunoregulatory effect of IL-10 expression, increase the content of probiotics and inhibit the growth of pathogenic bacteria in colitis rats, suggesting that F.prausnitzii combined with kukling can play a role in correcting intestinal flora disturbance.

【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R574.62

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