| Objective:This project is divided into two parts: clinical research and animal experimental research.Part of the clinical study evaluated the clinical efficacy of enteryu enema in the treatment of patients with ulcerative colitis(rectal type)with large intestine dampness-heat syndrome.Animal experiments were conducted to observe the effects of enteroyu enema prescription on the basic conditions of ulcerative colitis model rats,intestinal mucosal damage,intestinal status,the effect of Occludin and Claudin-1protein expression in the colon and the effect on the expression levels of Occluding,Claudin-1 and ZO-1 m RNA,and comprehensively observe the effect of enteroyu enema prescription on the mechanical barrier of intestinal mucosa,and theoretically analyze the relevant mechanism of enteroyu enema prescription in the treatment of ulcerative colitis(rectal type).Research methods:Clinical study: A total of 60 patients with rectal ulcerative colitis who met the diagnostic criteria and whose TCM syndrome was large intestine dampness-heat syndrome were selected as the research objects.According to the principle of random allocation,they were divided into the treatment group and the control group,both of which were 30 patients.The treatment group received enema treatment with enteryu enema prescription,and the control group received anal embolization treatment with mesalazine,the course of treatment was 4 weeks.Mayo score,TCM syndrome effect,and Baron score of rectal mucosa under the microscope were used as evaluation indexes before and after treatment.Animal experiment: 50 SD grade male rats were selected from experimental animal center of anhui medical university.They were 2 months old and weighed 200 ± 20 g.After 1 week,the rats were randomly divided into the normal group,the normal saline group,the mesalazine group,the medium dose enema group and the high dose enema group,with 10 rats in each group.In addition to the normal group,the other four groups of rats were intended to replicate the UC rat model with the TNBS/ anhydrous ethanol modified compound method.After the successful establishment of the model,rats in each group were treated with the corresponding drug enema for 4 weeks,and the changes of general signs and colonic mucosal injury index of rats in each group were observed.Immunohistochemistry was used to detect the expression changes of Occludin and claudin-1 in the colon tissues of rats,and PCR was used to detect the corresponding m RNA expression changes of Occludin,claudin-1 and zo-1.Results analysis:Results of the clinical study: a total of 58 patients were able to conduct statistical analysis,including 30 patients in the treatment group and 28 patients in the control group.After treatment,the Mayo score in both groups significantly decreased(P<0.05),indicating that both drugs were effective for the treatment of UC.The improvement of Mayo score and mucosal score under colonoscopy in the treatment group was better than that in the control group(P<0.05).The curative effect of TCM syndrome and mucosal score under colonoscopy were higher in the treatment group than in the control group(P<0.05).The results showed that the clinical efficacy of enteryu enema was better than that of mesalazine suppository.Animal experiment: after the success of the model,the rats in each model group were treated with brown and yellow watery stools or mushy stools,and the number of stools increased.Some of the stools were accompanied by mucous pus and blood stools.The food intake was significantly reduced and the weight was significantly reduced.Compared with the normal group,DAI scores of disease activity index were significantly increased in normal saline group,mesalazine group,medium dose enteryu enema group and high dose enteryu enema group(P<0.05).It shows that the UC rat model has been made.Comparison of the DAI score of disease activity index: compared with the normal group,the DAI score of disease activity index of rats in the normal saline group was significantly increased(P<0.05).Compared with the normal saline group,the DAI scores of the two groups with different concentrations of enteryu enema were significantly reduced and there were significant differences(P<0.05).There was no significant difference in DAI score between the mesalazine group and the medium dose enema group(P>0.05),while there was a significant difference between the high dose enema group and the medium dose enema group(P<0.05).Compared with the normal group,DAI scores of disease activity index were significantly increased in normal saline group,mesalazine group,medium dose enteryu enema group and medium dose enteryu enema group(P<0.05).It shows that the UC rat model has been made.Comparison of relative expression levels of Occludin and claudin-1 proteins in the colon tissues of rats: compared with the normal group,the expression levels of Occludin and claudin-1 proteins in the normal saline group were significantly decreased(P<0.05).After intervention,compared with the normal saline group,the expression of Occludin and claudin-1 proteins in each treatment group increased(P<0.05).The expression of Occludin and claudin-1 proteins increased in the high-dose enteroyu-enema group and the mesalazine suppository group(P<0.05).Compared with mesalazine group,there was no significant difference in the expression of Occludin and claudin-1 proteins in the high-dose enteroenema group(P>0.05).Comparison of relative expression levels of Occludin and Claudin-1 proteins in rat colon tissues: Compared with the normal group,the expressions of Occludin and Claudin-1 proteins in the colon tissues of the saline group rats were significantly reduced(P<0.05);after intervention treatment,Compared with the normal saline group,the expressions of Occludin and Claudin-1 proteins in the colon tissue of the three groups were increased(P<0.05).Compared with the medium-dose treatment group,the high-dose Changyu enema formula group and mesalazine The expressions of Occludin and Claudin-1 proteins in colon tissue of rats in the suppository group were increased(P<0.05).Compared with the mesalazine group,There was no statistical difference in expression(P>0.05).Comparison of transcriptional levels of Occludin,Claudin-1 and ZO-1 genes in the colon tissues of rats: compared with the normal group,m RNA expression levels of Occludin,Claudin-1 and ZO-1 genes in the normal saline group were significantly reduced(P<0.05).After intervention,the m RNA expression levels of Occludin,Claudin-1 and ZO-1 in each treatment group were significantly increased compared with the normal saline group(P<0.05).The expressions of Occludin,Claudin-1 and ZO-1 m RNA in the high-dose enteroyu-enema group and the mesalazine group increased(P<0.05).Compared with mesalazine group,the expression of Occludin,Claudin-1 and ZO-1 m RNA in the high-dose enteroyu-enema group was relatively increased(P<0.05).Conclusion:1.Enteryu enema prescription has a significant effect on patients with damp heat syndrome of ulcerative colitis(rectal type)large intestine,and is superior to the western drug mesalazine in improving Mayo score,colonoscopic mucosal score and clinical efficacy.2.Enteryu enema significantly improved the spirit,reaction,diet,feces and other general conditions of UC rats.3.Enteryu enema prescription has obvious effects on alleviating disease activity index and colonic mucosal injury index(CMDI)score in UC rats.4.The action mechanism of enteroyu-enema in the treatment of UC may be related to the increased expression of Occludin and claudin-1,as well as the increased expression of Occludin,claudin-1 and zo-1 m RNA.By maintaining the integrity and stability of TJ in UC rats,the mechanical barrier function of intestinal mucosa can be indirectly repaired to treat UC. |
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