The Efficacy Of QingChangHuaShi Method Treating Ulcerative Colitis And Experimental Study On TLR4/NF-κB Pathsway And Intestinal Mucosa Barrier

Objective Use QingChangHuaShi methods to treat ulcerative colitis, and then observe the efficacy and safety evaluation. Otherwise,To detect the effect of this method on the expression of NF-KB,TLR4,IL-8,occludin and claudin-1, and to explore the possible action mechanism through experiment.Methods Clinical part:According to the principle of randomization and contrast. The40cases diagnosed by colonoscopy with low-grade or middle-grade ulcerative colitis were divided into treatment group (20cases) and control group (20cases). Trratment group patients was given QingChangHuaShiFang(p.o.) and GuanChangFang (enem.).The control group was given Mesalazine (5-ASA)4g/d, The total period of treatment is six months. Observe the symptom disappeared-rates, the efficacy of TCM, Comprehensive effect,colonoscopy efficacy, the mucosal healing rates as well as the remission rate.Monitor the safety index. Experimental part:In vitro,The HT-29cell was induced to inflammation model by hTNF-a and LPS.HT-29cells were divided into6groups,as follows:normal control group,model control group,QingChangHuaShi Presciption high dose group, QingChangHuaShi Presciption middle dose group, QingChangHuaShi Presciption low dose group and SASP group. Use transwell to observe the macrophages migration,Use immune cell fluorescence method to detect NF-κB and TLR4protein, and use Elisa method to detect IL-8in the cells fluid. In vivo, Rat UC model induced by trinitrobenzene-sulfonic acid (TNBS) were randomly divided into normal group, model group, QingChangHuaShi Presciption group and SASP group. The rats were sacrificed to get their colonic mucosa and extract the whole-cell protein after10d of treatment. Western blot was performed to detect the protein levels of occludin and claudin-1.Results Clinical part: At the end of the treatment, the efficacy of colonscopy were better than the Mesalazine group(P﹤0.01). The two groups could improved abdominal pain,diarrhea, hematochezia,abdominal distension, anal burning,tenesmus, the efficacy of TCM, Comprehensive effect and ESR, but there were no statistical difference between the two groups. Also,There were no statistical difference in the mucosal healing rates (55.0%vs50.0%) and the remission rates (55.0%vs45.0%) between the two groups.There was no adverse events between the two groups too. Experimental part:In vitro, The macrophage in model group chemotactic increased, NF-κB activation into the nuclear apparently, TLR4protein expression and IL-8secretion increased.The role of High dose QingChangHuaShi Presciption to inhibit NF-κB activation is higher than SASP group (P﹤0.05).In vivo, QingChangHuaShiFang could lower both the pathology score and the general morphology score of colon. The protein expression of occludin and claudin-1in the model group were all lower than those in the normal group significantly. The protein expression of occludin and claudin-1in the QingChangHuaShi Presciption group were higher than that in the model group (P﹤0.05)Conclusion The results of clinical trial had shown that, QingChangHuaShi method was effective and safety in treating active ulcerative colitis of damp-heat syndrome,and were better than5-ASA in improving mucous membrane of colon. It was worthy to be popularized and make a further in-depth study. Experimental studies had shown that the therapy mechanism of QingChangHuaShiFang was involved in different degrees of intervention and regulation on the expression of NF-κB,TLR-4,IL-8,occludin and claudin-1. And then, It is may be through the above mechanism to suppress immune and reduce the inflammation.To achieve the purpose of treat ulcerative colitis.