Study On The Therapeutic Effects And Mechanism(s)of Tou Nong Sanon TNBS-induced Crohn's Disease In Rats

?Objective?To identify the effective components of the compound water decoction of TouNong San,and the therapeutic effect of TouNong San(TNS)on TNBS-induced crohn's disease in rats was evaluated by observing the pathological morphology of the colon,the levels of inflammatory factors in the intestines and serum of rats.On this basis,the mechanism(s)of treatment was explored,providing experimental basis for the clinical application of TouNong San in the treatment of inflammatory bowel disease.?Method?Pharmacodynamics aspect,this study adopts the weight only 200±20 g of SPF 60 SD male rats,raised in 22?±3?,humidity of 60%±10% of the animal room,fasting after 24 h with a dose of 100 mg/kg rectal enema dosing TNBS induced + ethanol mixed solution model of crohn's disease after building were randomly divided into normal control group,model group,positive drug group,and TouNong San low dose group(TNS-LD),medium dose group(TNS-MD),and high dose group(TNS-HD),and the treatment period is 7 days.The general conditions of the rats were observed daily and the changes in diet,water intake and body weight were recorded.On the 8th day,the serum was isolated for centrifugation and the supernatant was taken.The concentrations of TNF-?,IL-6,IL-1? and IL-17 were detected by ELISA.Colonic tissues were isolated and histologically graded at both macro and micro levels.And mRNA levels of TNF-?,IL-6,IL-1? and IL-17 in colonic tissues were detected by RT-qPCR,and IKK kinase,IKB protein and p65 in colonic tissues were detected by western blot.?Results?In this experiment,the experimental conditions of the fingerprint of TNS were optimized,and the main components in TNS were identified.Compared with the normal group,the weight,diet and water intake of the rats in each group were significantly decreased.HE staining and corresponding scoring showed that the positive drug group and the TNS groups could significantly improve the pathological morphology of the colon of crohn's disease rats,and reduce epithelial injury and submucosal inflammatory infiltration.Tissue and serum levels of inflammatory cytokines were significantly higher in the model group than in the normal group(P<0.01).The levels of inflammatory factors in tissues and serum were significantly improved in the positive drug group and the TNS groups(P<0.05).Mechanism aspect: compared with the normal group,p65 of model group were significantly increased in the nucleus,IKB protein significantly decreased,IKK kinase in the cells increased significantly,this has proved its inflammatory pathways are activated in the process of experiment.The p65 levels of TNS groups in the cell nucleus and cells IKK kinase decreased obviously,IKB protein level increased significantly.?Conclusion?(1)TNS can significantly inhibit the intestinal inflammation of TNBS-induced crohn's disease rats,improve their histopathological morphology,and reduce the levels of inflammatory factors in tissues and serum.(2)the improvement of inflammation in rats with crohn's disease may be related to its activity of inhibiting the NF-?B pathway.