| Objective:Based on protein chip technology and enrichment analysis to screen relevant targets,the possible mechanism of Ding's Herb Enema Prescription(DHEP)to improve the intestinal inflammatory response in Ulcerative Colitis(UC)rats by regulating multiple cytokine targets was studied.Methods:48 rats were randomly divided into 6 groups,8/group,which were blank group,model group,positive group(Mesalazine enema group),Ding's Herb Enema Prescription solution high-dose group,Ding's ulcerated enema fluid.Dose group,Ding's Herb Enema Prescription liquid low dose group.Except for the blank group,the other five groups were induced with Dextran Sulfate Sodium(DSS)in a rat UC model.After successful modeling,each group was administered enema,once a day for 14 days,during which the general state of the rats was observed,including mental activity,dietary status,fecal nature,blood in the stool,change in body weight to calculate the weight gain rate,etc.Rat Disease Activity Index(DAI)was evaluated.One day after discontinuation of the drug,the rats were sacrificed and the colonic mucosa of the rats was removed.The protein changes of the intestinal inflammation were observed by protein microarray and the relevant targets of each group were screened.Result:1.After the rats were modeled,symptoms such as diarrhea,mucopurulence,and weight loss appeared.The DAI score was higher than that of blank control(P<0.01).After treatment,there was a statistically significant difference between the blank group and the model group(P<0.01).The treatment effect of the middle and high dose Ding's Herb Enema Prescription group was similar to that of positive group,and different concentrations of Ding's Herb Enema Prescription were observed.The comparison between the liquid and the model group showed that the low-medium-high group had therapeutic effects(P<0.01;P<0.05).2.Through the naked eye,the rats in the model group had erythema on the colon wall,normal texture of mucosal blood vessels disappeared,intestinal wall edema,diffuse hemorrhage,increased brittleness,thickened and dulled septum of colon pouch,accompanied by erosion and ulceration.The score was higher than the blank control group(P<0.01).The colonic mucosa of the positive group and Ding's Herb Enema Prescription solution only showed colonic erythema,edema,and mild erosion,but there was no obvious ulcer.The score was lower than that of the model group(P<0.01;P<0.05).3.After light microscope observation,in the model group rats,inflammatory lesions were confined to the mucosa and submucosa,accompanied by crypt abscess formation,vascular congestion,and decreased mucus produced by crypt epithelial cells,and there were a large number of neutral particles.The score of cells was significantly higher than that of blank control group(P<0.01).There was no obvious ulceration in the colon mucosa of the positive group and Ding's Herb Enema Prescription group.The inflammatory cell infiltration was reduced and the score was significantly lower than that of the model group(P<0.01).4.GSR-CAA-67 antibody protein chip technology was used to select differentially expressed proteins.IFN-?,erythropoietin(EPO),TIMP-2,TIM-1,IL-6,TIMP-1,TNF-?,and IL-22 were screened by comparing the model group with the blank control group.Proteins,the differences were statistically significant(P<0.05).In addition,Ding's Herb Enema Prescription group and mesalazine group had significant inhibitory effects on IL-6 and TIMP-1(P<0.05).Ding's Herb Enema Prescription group had TNF-a and IL-22 levels.Inhibitory effect(P<0.05),positive group had no inhibitory effect on TNF-?,IL-22(P>0.05).Conclusion:1.Ding's Herb Enema Prescription solution has a certain therapeutic effect on UC rats,relieves symptoms such as diarrhea,mucopurulence,and weight loss,and promotes the healing of intestinal mucosal ulcers.2.Ding's Herb Enema Prescription can improve the intestinal inflammation of UC rats by regulating multiple cytokines and inflammatory mediators and achieve the purpose of treating UC. |
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