| Objective The pathological changes of rats in different concentration of probiotics in the treatment of ulcerative colitis in rats, Changes in the expression of inflammatory factors and the changes of intestinal flora to exploration the optimal dose of probiotics in the treatment of ulcerative colitis and intestinal microflora, To provide theoretical basis for clinical diagnosis and treatment of UC.Methods1. The experimental group:39 rats were randomly divided into, The blank control group, DSS model group, saline group, low concentration group, the concentration of treatment group, high concentration group, heat inactivated treatment group.2. The object of study:Rat colitis and colon tissue and feces.3. The test content and method:Observe the general condition of the rats during the experimental period, activity, diet, body weight monitoring, Observe the stool occult blood and have no observation, e. anatomy of rat after the gross changes and the measurement of length, General damage on colonic morphology score. The rat colon tissue HE staining score and the testing level and cytokine by immunohistochemical method. firstly feces smear, gram stain and bacterial flora analysis were carried out; and then bacteria strains were be cultured, isolated and identified from feces by traditional bacteriology method, include aerobic and anaerobic culture; finally,6 bacterial toxin genes were be detected by real-time fluorescent quantified polymerase chain reaction (RT-PCR) method, including the heat-labile enter toxin gene(LT) of enterotoxigenic escherichia coli, toxin vira gene of enteroinvatsive escherichia coli, enterotoxin bft gene of bacteroides fragilis, exotoxin cpa gene of clostridium perfringens, and toxin A gene and toxin B gene of clostridium difficile.4. Statistical processed:data analysis was performed using SPSS 19.0.Results1、At the end of modeling, the DSS model group rats, DIA score of 6.6+1.1402, significantly higher than the control group; the pathological score was 8.75+0.5, significantly higher than the control group.2、Did not receive probiotic gastric gavage of DSS rats in the general condition of the poor, some rats appeared mucopurulent bloody stool or bloody stool, the blank control group rats have no abnormal performance, the general condition of rats in the treatment group was the blank control group and model group. The DAI value of the treatment group with the prolongation of treatment time showed a downward trend, the end of the treatment, the treatment group was significantly lower than that of DSS model group DAI score higher than that of the control group.3、 After probiotics treatment, each treatment group rats colitis lesions compared with DSS model group reduced significantly, and the middle dose group and high dose treatment of colon injury group rats were reduced more significantly. And after the probiotic treatment, reduce the pathological score of colon mucosa of rats in all treatment groups compared with the DSS model group rats showed different degrees of reduction of colonic mucosal erosion, ulcer and inflammation recess and crypt abscess, The lesion range is narrow, the extent of damage of intestinal epithelial loss, only a part of glandular epithelial cells necrosis, infiltration of inflammatory cells also decreased.4、Immunohistochemical detection of inflammatory cytokines, DSS made module rats colonic inflammatory factor INF-α、IL-6 the highest level anti-inflammatory factor TGF-P IL-10 level was the lowest. The probiotic treatment, the treatment group of inflammatory factor TNF-a IL-6 expression compared with DSS model group decreased obviously, anti-inflammatory cytokine of TGF-P IL-10 levels were significantly increased. And the level of anti-inflammatory cytokines probiotics high concentration increased more obviously.5、Fecal microbiota smear results show that each treatment group flora was lower than that of DSS group. The treatment group of flora imbalance degree is lower than DSS model group, significantly higher than the control group. The results showed that the fecal culture and identification of the pathogens, treatment group was lower than that of DSS growth model, the growth rate is higher than the DSS probiotic advantage model.Conclusion1、DSS modeling method can be successfully induced rat colitis model, similar to human UC. And the method is simple and easy, good repeatability, high safety factor.2、Different doses of VSL#3 to alleviate the symptoms of UC in colitis rats to different degrees, probiotics, high dose of VSL#3 (10.8×1011CFU Kg/day) effect is most obvious, middle dose (1.08×1011CFU Kg/day) of low dose (0.108×1011CFU Kg/day) and poor effect of heat inactivated probiotics.3、VSL#3 can repair damage in inflammatory tissue of colitis rats, alleviate the symptoms of inflammation in rats, reduce the pro-inflammatory cytokines in colon tissue the content of IL-6、TNF-α, Inhibition of the secretion of pro-inflammatory cytokines; Improve the content of anti-inflammatory factor TGF-β、IL-10, Blocking the inflammatory response, shorten the duration of inflammation.4、VSI#3 can reduce the intestinal dysbacteriosis severity to a certain extent, reduce the flora imbalance rate. Increase the Bifidobacterium, Bacteroides bacteria ratio. Virulence genes to reduce intestinal colitis in rats with invasive Escherichia coli plasmid carrying rate. |
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