| Objective:1.The pathogeny and pathogenesis of ulcerative colitis were complicated and it has a wide variety of syndrome types.Most of physicians treated it based on syndrome differentiation and there was a good curative effect in clinical so far.However,there is a limitation of clinical promotion,because syndrome-types and treating methods of ulcerative colitis are too scattered,and even we have not make a further discussion of its treatment and medicine regularity.Now we analyze the pathogenesis and treatment of ulcerative colitis in recent years by frequency analysis method to discuss the common pathogenesis and treatment of ulcerative colitis,in order to provide a documentary basis for clinical choose prescriptions and in-depth study.2.Sishen wan is an effective prescription to ulcerative colitis of yang deficiency of spleen and kidney.But the mechanism of action for treatment of ulcerative colitis is completely unknown.By observing the impact of ulcerative colitis of yang deficiency of spleen and kidney model rats’ serum and colonic IFN-γ,TGF-beta 1 factor content and colon tissue of NOD2,gene of SIGIRR and protein expression which take Sishen wan,then discuss the molecular mechanism of ulcerative colitis of yang deficiency of spleen and kidney treated by Sishen wan,in order to provide experimental basis for further application and development of the prescription.Methods:We use Senna irrigation stomach and hydrocortisone intraperitoneal injection +TNBS/ethanol mixed suspension enema to make rat models of ulcer colitis of yang deficiency of spleen and kidney.We evaluated the rat models through the rats’ general state, colon injury,pathological changes and so on.Then the experimental rats were randomly divided into blank group, model group,SASP group, Sishen wan low,medium and high dose group.Give SASP group 0.36g/kg/d drug dose gavage and Sishen wan low,medium and high dose groups dose gavage of 2.5,5,10g/kg/d. Blank group,model group were treated with equal volume of distilled water,continuous treatment of 21 d.Use ELISA to detect the content changes of rattissues and serum IFN-γ and TGF beta 1 factor;use RT-q PCR and Western blot method to detect the changes of rat colon NOD2,SIGIRR gene and protein expression.Discuss the expression changes of IFN-γ and TGF beta 1,NOD2, SIGIRR and the intervention effect of Sishen wan.Results:1.General state:rats in blank group were clean and smooth,good spirit,regular diet,normal defecate and there was no significant changes in the model and treatment period.Model group rats have unclean hair,poor spirit, sleepy and inactive,watery stool,onyx with lighter color,cold tail,atony arch,clinging to cluster,decreasing in eating and drinking after building period.Because itself has a certain repair cycle,in the 7th day after building,syndrome was most severe,and they didn’t have a certain improvement until 14 th day,while there was a most obvious improvement in the 21 th day.But compared with other treatment groups,the repairing effect was not very well.Through a period treatment with different drugs and different doses,the rats in each group had obtained different repairing levels.SASP group rats’ fur get cleaner,flexibility recovered slightly,has normal spirit,sensitive activity and normal defecate,only a few have watery stool.Rats in Sishen wan group:luster fur,sensitive reaction,good spirit, closing to the normal diet,granular stool.Among them,the middle dose group recovered best,there was no significant difference with the blank group.2.Gross morphology:colonic mucosa of model rats group had obvious congestion, edema,ulcers,bowel wall thickening,intestinal shortened and etc.Which confirmed the model were made correct(P<0.01).After three doses of Sishen wan and SASP treatment,rat colon mucosa destruction were improved,but it was different in each group.Wherein,in SASP group,Sishen wan high-dose group,there is only a small amount of congestion and edema in rats mucous membrane medium dose group had almost returned to normal form.Sishen wan group,SASP group contrasted with model control group,colon injury score all decreased(P<0.01).3.Pathological changes:in the blank group,the colon tissue was clear,the mucosa was complete,no ulcer,inflammation and so on,no obvious abnormality was found;in the model group,the structure of colon was not clear,inflammatory cell infiltration,depth of myometrial,ulcer formation,colon full layer thicknesss;tructure is not clear,a large number ofinflammatory cell infiltration,depth of myometrial,ulceration,colon full layer thickness;in SASP group,low dose of Chinese medicine group and the high dose Chinese traditional medicine group,the epithelium had different degree of recovery,muscle layer had slightly repair,but fault was still existing,in colon there were even a few inflammatory cells;in Chinese medicine dose group,colon structure is relatively complete,clear,close to normal tissue structure.4.Use ELISA to detect the content of IFN-γ and TGF-β1:Compared with the blank group,either in serum or colon of model group rats,the contents of IFN-γ were increased,meanwhile,the contents of TGF-β1 were decreased(P<0.01).Compared with the model group,either in serum or colon of treatment group rats,the contents of IFN-γ were decreased,while the contents of TGF-β1 were increased(P<0.05, P<0.01).5.Use RT-q PCR method to detect the expressions of NOD2 and SIGIRR m RNA:compared with the blank group,the expressions of m RNA NOD2 in model group rats were increased(P<0.01),furthermore,the expressions of m RNA SIGIRR were decreased(P<0.01);compared with the model group,the expressions of m RNA NOD2 in the treatment group were decreased(P<0.05),while the expressions of m RNA SIGIRR were increased(P<0.01, P<0.05).6.Use Western Blot method to detect the expressions of NOD2 and SIGIRR:compared with the blank group,the expressions of NOD2 in model group rats were significantly increased(P<0.01),in the contrary,the expressions of SIGIRR were decreased(P<0.01);compared with the model group,the expressions of NOD2 in rat colons of treatmentl group were decreased(P<0.01),while the expressions of SIGIRR increased(P<0.05).Conclusion:1.Using senna orally + hydrocortisone injection intraperitoneal injection + 2,4,6-trinitrobenzene sulfonic acid(TNBS)/ethanol enema can successfully establish rat models of ulcerative colitis of yang deficiency of spleen and kidney.2.Yang deficiency of spleen and kidney is an important and common UC evidence,wenshenjianpi treat is an important therapy for the treatment of UC.3.The macroscopic observation and pathological detection of rat colons tissue formsuggest that all of the Sishen wan low,medium and high dose have different levels of repair action on the lesion of colonic mucosa.4.Sishen wan makes decreased expressions of proinflammatory cytokines IFN-γ,NOD2,etc.in ulcerative colitis model rats serum and colons,and makes increased expressions of anti inhibiting inflammatory cytokine TGF-β1,SIGIRR and etc.Its mechanism of action may be through downregulation of proinflammatory factors and upregulation of anti inflammatory factors to keep balance of immune regulation,and promote mucosal repairing and ulcer healing,in order to achieve the direction of treating ulcerative colitis. |
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