| Backgroud:Ulcerative colitis(UC)is a chronic inflammatory disease characterized by long-term unpredictability and uncertainty,which can often trigger anxiety and depression in patients.Recent clinical studies have shown that depression is more common in patients with ulcerative colitis in the years prior to diagnosis,and that depression can exacerbate the progression of ulcerative colitis.The cause is unknown and may be closely related to gut microbiota.The Traditional Chinese medicine,Abelmoschus Manihot,as the "essential medicine for sores",is often used in the treatment of boils caused by carbuncle,rax,and skullcap.In the clinical practice of our department,Abelmoschus Manihot,is a commonly used Chinese medicine for ulcerative colitis.TFA is effective compound extracted from Abelmoschus Manihot and have significant therapeutic effects on Crohn's disease,chronic glomerulonephritis and poststroke depression.Therefore,it is speculated that the total flavone of Abelmoschus manihot can improve ulcerative colitis in depression state by regulating gut microbiota.Objective:To observe whether patients with clinical depression can disorder the gut microbiota of patients with ulcerative colitis,as well as the effect of the total flavone of Abelmoschus manihot on patients with ulcerative colitis in depression state,and to explore its internal mechanism and target from the direction of gut microbiota,so as to provide experimental basis for clinical application of the total flavone of Abelmoschus manihot.Methods:Clinical TrialsThis study included 72 active UC patients(36 with depression and 36 without depression).16srRNA method was used to detect gut microbiota of all subjects to identify gut microbiota characteristics related to UC depression.Experiments on animalsExperiment ?80 male C57BL/6J mice were randomly divided into 8 groups,the CK(Blank control)group,the CS(Chronic stress)group,TFA(low,L)+CS group,TFA(high,H)+CS group,DSS group,CS+DSS group,TFA(L)+CS+DSS group and TFA(H)+CS+DSS group.Depression model was established by using chronic restraint stress.After chronic stress modeling,the mice were subjected to behavioral tests(open-field,suspended tail test and forced swimming)to evaluate their depression-like performance.Depression UC mice were given 2.5%DSS solution after 30 days of chronic restraint stress for 7 days.During the experiment,TFA were administered daily by gavage.At the end of the experiment,mice were euthanized and cecal contents and colon tissues were collected.Colon inflammation in mice was identified by measuring colon length,assessing Disease Activity Index(DAI)and colon pathology,and detecting colon IL-6,IL-1? and TNF-? mRNA expression by qPCR.16S rRNA was used to detect the effect of TFA treatment on gut microbiota in mice.MUC2,KLF4 and Z0-1 positive cell staining,PAS-AB staining and MUC2 KLF4 ZO-1 mRNA expression were used to evaluate the repair effect of TFA on intestinal barrier function.Experiment ?Fecal Microbiota Transplantation(FMT)was used to further investigate whether the efficacy of TFA in treating depression UC was mediated by gut microbiota.First,90 C57BL/6J male mice were randomly divided into 30 donor groups and 60 recipient groups.The donor group was divided into CK group,CS group and TFA(L)+CS group.The donor group was modeled as experiment 1.After the modeling,the mice were euthanized,and the cecal contents were made into bacterial liquid.The mice in the recipient group were given mixed antibiotics for antibiotic cleaning.After the cleaning,the bacteria solution of recipient mice was transplanted into recipient mice,and then colony colonization was carried out,in which 30 recipient mice were subjected to behavioral detection and the other 30 recipient mice were given 2.5%DSS solution to drink freely for 7 days after colony colonization.After the experiment,the recipient mice were euthanized and the cecal contents and colon tissue were collected.The colonic injury,gut microbiota and intestinal barrier function of the recipient mice were evaluated as experiment?.Results1.Depression can alter the diversity and composition of gut microbiota in patients with ulcerative colitis.2.TFA can improve depression-like behavior in chronic stress mice,increase the range of activity in open field test,and reduce the immobility time of mice in tail suspension test and forced swim test.3.TFA improved colitis with chronic stress,increased colonic length,improved DAI score,HE staining score,and mRNA expression of IL-6,IL-1? and TNF-? in mice.4.TFA improved gut microbiota of mice,provide diversity of flora,improve beneficial bacteria,and reduce harmful bacteria.5.TFA can improve the intestinal barrier function of mice,increase the number of PAS-AB goblet cells,MUC2,KLF4,ZO-1 positive cells and increase the expression of intestinal barrier of MUC2,KLF4 and ZO-1 mRNA.6.TFA improved depression-like behaviors of mice by gut microbiota,increase the range of activity in open field test,and reduce the immobility time of mice in tail suspension test and forced swim test.7.TFA improved the intestinal barrier function of mice by FMT,increase the number of PAS-AB goblet cells,MUC2,KLF4 and ZO-1 positive cells,and improve the expression of intestinal barrier of MUC2,KLF4 and ZO-1 mRNA.8.TFA improved gut microbiota of mice by FMT,provide diversity of flora,improve beneficial bacteria,and reduce harmful bacteria9.TFA improved colitis with chronic stress by FMT,increased colonic length,improved DAI score,HE staining score,and mRNA expression of IL-6,IL-1 ? and TNF-?.10.The abundance of Alistipes decreased and the difference increased in the pathogenic process of chronic stress and in the treatment of TFA.ConclusionClinically,depression interferes with the gut microflora of patients with ulcerative colitis.Animal studies have shown that TFA can alleviate the colitis of mice under the state of depression through the intestinal mucosal barrier function mediated by gut microflora,and Alistipes may play an important role in the pathogenesis of depression and the treatment of TFA. |
PDF Full Text Request