Construction Of A Risk Prediction Model For Fecal Incontinence In CD And The Amelioration Of TNBS-induced Colonic Fibrosis In Mice With Total Flavone Of Abelmoschus Manihot

BackgroundCrohn’s disease is a non-specific inflammatory disease of the intestine that affects the entire tract of the digestive tract from the mouth to the anal.The disease is segmented and penetrates the entire intestinal wall.Fecal incontinence(FI)is a frequent complication of CD,defined by the involuntary loss of liquid or solid.Because of social stigma and embarrassment,FI is an underdiagnosed symptom in IBD,which leads to the serious neglect of the problem,bringing insidious harm to patients.However,most of the previous related studies have focused on the analysis of related factors and the impact on the quality of life.There are no tools available for clinicians to effectively predict FI occurrence,making early intervention difficult.AimTo explore the independent risk factors influencing the occurrence of FI in patients with CD,constract a FI prediction model,and verify the model in internal data sets and prospective cohort.MethodsThis was a multicenter,combined prospective and retrospective observational study.In the retrospective study,patients with CD treated in Jiangsu Hospital of Traditional Chinese Medicine from June 2016 to April 2021 were collected.Collect relevant data,disease information,etc.,and record the patient’s Wexner score.Patients were divided into FI group(Wexner≥5)and non-FI group(Wexner<5)according to Wexner score.Univariate analysis was used to initially screen out statistically significant indicators.A bivariate logistic regression analysis was used to construct a prediction model.And the Nomogram was visualized.Plot ROC curves to evaluate model performance and perform internal validation of predictive models.In a prospective study,CD patients who visited five IBD clinics from 2021.06 to 2021.10 were collected with the same inclusion and exclusion criteria.According to whether the patients developed FI,they were divided into FI group and non-FI group,the risk factors of the two groups were compared,and the AUC value representing the model discrimination and the HosmerLemeshow test of consistency were calculated.Result1.Construction and validation of a risk predictive model for FI in CD patients.A total of 461 CD patients were included,and the overall incidence of FI was 28.8%(133/461).Using the bootstrap resampling method,the overall data were randomly divid ed into training set(368 people)and validation set(93 people)according to 8:2.The trai ning set was used to construct the prediction model,whereas the validation set was used to evaluate the efficiency of the prediction model.According to the presence or absence of FI,the training set samples were divided into FI group and non-FI group.After comparison,it was found that FI was related to BMI,history of non-fistulae perianal disease,number of loose stools in the past week,current anal fistula,PDAI score in active perianal dise ase>4,and CDAI score in active disease>150(P<0.05).Multivariate regression analysis showed that the history of surgery for non-fistula perianal disease(OR=2.1,95%CI 1.48-5.67),the numbe of loose stools in the last week was 1-14(OR=2.05,95%CI 1.16-3.98),The number of loose stools in the last week≥14(OR=2.78,95%CI 0.995-4.35)and active perianal disease(OR=2.57,95%CI 1.566-3.4.899)were independent of fecal inconti nence in CD patients risk factors.Increased BMI was a protective factor for FI(OR=0.91,95%CI 0.822-0.976).The FI prediction model was constructed based on the AIC princ iple,and the risk prediction nomogram was established.Internal validation of the model showed an AUC of 0.797,showing great accuracy for the model.2.External validation of fecal incontinence risk prediction model in CD patients.A total of 225 patients were prospectively enrolled,and the overall incontinence rate was 4.9%(11/225).The patients were divided into the FI group and the non-FI group.The comparison found that the two groups had complex anal fistula,a history of perianal abscess,the number of perianal operations ≥ 3 times,the current anal fistula,and the active perianal disease.The distribution of patients was statistically different(P<0.05).The AUC value of the prediction model in the validation population was 0.7795,and the Hosmer-Lemeshow test was0.826,showed that there was good consistency between the predicted FI of the response model and the actual probability.ConclusionThis study developed a predictive model for the risk of FI in CD patients.The model included five predictors:non-fistula perianal disease surgical history,the number of loose stools in the last week,the degree of perianal disease activity,and BMI.It has been verified that the model has good accuracy and discrimination,and can be used as an important prediction tool for the risk of FI in CD patients,providing patients with early prevention and timely intervention.BackgroundCrohn’s disease(CD)is a chronic and idiopathic inflammatory disease that can involve any part of the digestive tract.Although the clinical manifestations and natural course of the disease vary in CD patients,transmural inflammation leading to fibrosis of the intestinal wall,intestinal stenosis,and ultimately intestinal obstruction are the characteristic clinical manifestations.Surgery remains the major available strategy in inflammatory bowel disease(IBD)fibrotic strictures because no available drugs have sufficient prevention and treatment in this complication.Abelmoschus manihot L.Medic is a traditional Chinese medicine,commonly used to treat diseases such as surgical sores.Clinical application in the treatment of IBD has achieved good results.Total flavone of Abelmoschus manihot L.Medic(TFA)is the main flavonoid compound extracted from Abelmoschus manihot.Modem pharmacological studies have shown that TFA has anti-inflammatory,inhibiting epithelial-mesenchymal transition mechanism,and inhibiting renal fibrosis.Therefore,it is speculated that TFA can interfere with intestinal fibrosis in CD.ObjectiveThis study aimed to evaluate the efficacy of the total flavone of Abelmoschus mamhot L.Medic(TFA)on the development of colonic fibrosis in mice and its possible mechanism.MethodsChronic colonic inflammation-associated fibrosis was induced in BALB/c mice by 4 weekly repeated administration with 2,4,6-trinitrobenzene sulfonic acid(TNBS).Animals were assigned to five groups: ethanol group,saline group,TNBS group,TFA in modeling group,TFA after modeling group.The colon was removed after sacrificed and assessed by macroscopic,histological and immunohistochemical analyses.Inflammatory cytokines in serum and tissue was examined with ELISA.Extracellular matrix(ECM)deposition in the colon was examined with image analysis of Masson staining and Verhoeff’s Van Gieson(EVG)Staining.Total levels of colla2,col3a2,TGF-β,α-SMA,Vimentin, MMP2,MMP9 and TIMP-1 were measured by reverse transcription-quantitative polymerase chain reaction or Western blot analysis,respectively.Results1.Compared with the model group,oral administration of TFA attenuated body weight loss,reduced colon length shortening,lowered the morphological damage index score,and notably ameliorated the inflammatory response.2.Masson staining and EVG staining showed that after repeated administration of TNBS,intestinal fibrosis in the colon of mice was significantly increased.The content of collagen fiber in the TFA intervention group was significantly lower than that in the modeling group3.Elisa experiment showed that compared with model group,TFA downregulated proinflammatory cytokines IL-6,IL-17,TNF-α,IFN-γ productions,and increased the levels of anti-inflammatory cytokine IL-10 and TGF-β.4.TFA significantly reduced the expression of fibrosis-related collagens colla2,col3a2 and hydroxyproline,the main component of collagen,and reduced the pro-fibrotic component TGF-β and markers of myofibroblast phenotype in a TNBS-induced colonic fibrosis model.Levels of α-SMA,a mesenchymal cell maker vimentin;decreased expression of TIMP-1 and increased levels of MMPs to degrade extracellular matrix deposition.ConclusionTogether,we herein provide the evidence to support that TFA may restore the imbalance of Thl7/Treg and decrease the generation of ECM.This may be a potential mechanism by which TFA protects the intestine under inflammatory conditions and acts as a therapeutic agent for the treatment of intestinal fibrosis in Crohn’s disease.