Jiaotai Wan Alleviate Gut Microbiota-mediated Intestinal Mucosal Barrier Damage In The Stress Mice Following Colitis

Clinical and experimental evidence indicates that stress increases disease activity during ulcerative colitis,attributed to gut microbiota.Jiaotai Wan(JTW)has a long history of treating psychological stress.However,effect of JTW on concurrent stress and colitis is unclear.Firstly,Stress was induced by chronic restraint stress to imitate psychological stress and colitis was induced by drinking dextran sulfate sodium(DSS)to imitate ulcerative colitis,which explored that JTW could regulate alteration of gut microbiota induced by stress,thereby improving the colitis.Secondly,we built fecal microbiota transplantation(FMT)to determine whether JTW could improve progression of colitis by intestinal barrier function repairment driven by gut microbiota.Experiment 1Methods80 male C57BL/6J mice were randomized into eight group;Normal group(n=10),Stress group(n=10),JTW(high dose)+stress group(n=10),JTW(low dose)+stress group(n=10),DSS group(n=10),Stress+DSS group(n=10),JTW(high dose)+stress+DSS group(n=10),JTW(low dose)+stress+DSS group(n=10).Stress model was conducted by continuely restraint for 30 days.Subsequently,the colitis model was established by drinking 2.5%DSS solution for 7 days.JTW were administered from chronic restraint stress until the end.At the end of the experiment,all mice were sacrificed.The colon tissues and cecal contents were collected for the next detection analysis.The progression of colitis were assessed by colon length,the disease activity index(DAI)and histological scores.The intestinal mucosal barrier were assessed by PAS-AB staining and MUC2 immunohistochemical staining.qPCR were used to observe the differences in the expression of KLF4?MUC2?IL-1??IL-6?TNF-? in mice colon tissues.Cecal contents were profiled using 16S rRNA sequencing.ResultsChronic restraint stress induced depression-like behavior and exacerbated the development of colitis in mice.JTW administration could effectively relieve depressive-like behaviours.Besides,stress-induced intestinal mucosal barrier damage was repaired by intervention of JTW.Colon tissue mRNA expression of KLF4 and MUC2 decreased.High dose intervention of JTW could relieve intestinal injury on the concurrent stress and colitis,which was evidenced by reduced colon length shortening,DAI scores and histopathology scores.Colon tissue mRNA expression of inflammatory cytokines(IL-1??IL-6?TNF-?)decreased as a result of JTW administration.Analysis of 16S rRNA sequences demonstrated that JTW induced significant gut microbiota alterations,which included increased relative abundance of Akkermansia and decrceased Escherichia-Shigella at the genus le-vel.Experiment 2MethodsWe used a model of fecal microbiota transplantation(FMT)to further determine whether effect of JTW on concurrent stress and colitis were mediated by gut microbiota.125 male C57BL/6J mice were randomized into donor group and recipient group.Donor group included normal group,stress group,JTW group.The mixed antibiotic-induced recipient mice with FMT of microbiota from different donor mice were conducted.At the end of microbiota colonization,part of the mice were performed behaviorally test and sacrificed,and another part were free to drink 2.5%DSS solution for 7 days to establish colitis.At the end of the experiment,colon tissues and cecal contents of mice were collected.The progression of colitis,intestinal mucosal barrier damage and gut microbiota alterations were assessed by referring to Experiment 1.ResultsThe results showed that intestinal mucosal barrier damage and colon tissue mRNA expression of inflammatory cytokines(IL-1??IL-6?TNF-?)decreased in the JTW recipients.Meanwhile,gut microbiota of JTW recipients were significantly changed,which included increasing relative abundance of Akkermansia and decreasing Escherichia-Shigella at the genus level.Notably,stress recipients with live Akkermansia muciniphila reduced depressive-like behaviours,intestinal mucosal barrier and colon tissue mRNA expression of inflammatory cytokines.ConclusionsOur results indicate that JTW could alleviate gut microbiota-mediated intestinal mucosal barrier damage in the stress mice following colitis.Besides,JTW may be used as prebiotics of Akkermansia to treat concurrent stress and colitis.