Clostridium Butyricum Protect And Relieve The Visceral Hypersensitivity In Mice Induced By Citrobacter Rodentium Infection With Chronic Stress

Part ?:Clostridium butyricum relieve the visceral hypersensitivity in mice induced by Citrobacter rodentium infection with chronic stressBackground:Visceral hypersensitivity is a common symptom in patients with postinfectious irritable bowel syndrome(PI-IBS),and change of the microbiota is a vital etiological factor of it.Clostridium butyricum(C.butyricum)is one of the probiotics which is reported as the active components in the treatment of IBS,especially IBS with diarrhea.Citrobacter rodentium(C.rodentium)is an enteropathogenic bacteria which can produce self-limiting colitis in mice,which have been used to produce a PI-IBSlike mice model.Whether C.butyricum could influence the visceral hypersensitivity and gut microbiota of PI-IBS is still unknown.Our study aimed to examine whether the intervention of C.butyricum or antibiotics could affect the etiology of visceral hypersensitivity.Methods:(1)C57BL/6 male mice were gavaged with the C.rodentium to induce a infective colitis.The C.butyricum and antibiotic compound were used to intervene the infected mice 3 days later.A 9-days chronic water avoidance stress(WAS)process was implemented to help induce the visceral hypersensitivity.(2)The abdominal withdrawal reflex(AWR)score was detected to show the visceral hypersensitivity of different mice groups.(3)The HE staining was conducted in the colonic histology specimens of the six C.rodentium infected mice,and we chose to sacrifice the mice at D0,D5,D21 after the infection.(4)On the 7th,14th,and 30th days after infection,mice feces were collected and high-throughput sequencing was carried out to analyze their gut microbiota.Results:(1)Combined post the C.rodentium infection plus chronic stress(WAS)could induce the visceral hypersensitivity in mice.(2)Treatment of the C.butyricum after the C.rodentium infection could relieve visceral hypersensitivity of mice,while no difference was observed in the antibiotic treatment group.(3)The colonic inflammation pathological changes of the infectious group had recovered to normal within the assessment of the histopathological score at Day 21 after C.rodentium infection.(4)The gut microbiota diversity of C.rodentium infected mice was similar to the noinfected mice,while there showed different microbial communities' structure between them.The Shannon and Chao indexes significantly decreased in the antibiotic treatment group compared to other groups at 7th,14th,and 30th post-infection days,while treatment of C.butyricum could maintain the indexes within normal range.At day 14 after infection,the structure of microbiota headed towards normality after the C.butyricum treatment.After the WAS,the Shannon and Chao indexes of the control group decreased and the structure of microbiota changed.The C.butyricum treatment could prevent these changes of the gut microbiota induced by WAS.Conclusion:C.butyricum could relieve the visceral hypersensitivity in mice induced by C.rodentium infection plus chronic stress.It could also remodel the microbiota change caused by the infection and chronic stress.It may be a more effective treatment strategy for PI-IBS than antibiotics.Part ?:Clostridium butyricum protect the genaration of visceral hypersensitivity in mice induced by Citrobacter rodentium infection with chronic stressBackground and objectives:Irritable bowel syndrome(IBS)is one of the most common functional gastrointestinal disease in worldwide,characterized as the disorder of gastrointestinal movement and the visceral hypersensitivity.Alteration of gut microbiota is one of the most important etiology of IBS.There is one kind of probiotics named Clostridium butyricum(C.butyricum)has been used in the treatment of IBS,especially the diarrhea-predominate IBS.Post-infectious irritable bowel syndrome(PI-IBS)is one subtype of IBS,which means the patients which show the IBS-like symptoms after recovered from acute gastroenteritis.There was no specific study has examined the role of C.butyricum on the prevention for PI-IBS.In our previous study about C.butyricum,we found that this probiotic could relieve the visceral hypersensitivity of PI-IBS-like model mice,and gut microbiota play an important role on it.Whether the C.butyricum could influence the visceral hypersensitivity and gut microbiota of PI-IBS is still unknown.Our study aimed to examine whether the advanced interfere treatment of C.butyricum or antibiotics could affect the generation of visceral hypersensitivity,and how did the gut microbiota change during this process.Methods:(1)C57BL/6 male mice were gavaged with the C.rodentium to induce a infective colitis.The C.butyricum and antibiotic compound were early used to intervene the infected mice 7 days before infection.A 9-days chronic water avoidance stress(WAS)process was implemented to help induce the visceral hypersensitivity.(2)The abdominal withdrawal reflex(AWR)score was assayed to indicate the visceral hypersensitivity of different groups.(3)On the 7th,14th,and 30th days after the infection,mice feces were collected and high-throughput sequencing was carried out to analyze their gut microbiota.Results:Combined post the C.rodentium infection plus chronic stress(WAS)could induce the visceral hypersensitivity in mice.Treatment of the C.butyricum before C.rodentium infection could relieve visceral hypersensitivity of mice,while no difference was observed in the antibiotic treatment group.The gut microbiota diversity of C.rodentium infected mice was similar to the no-infected mice,while there showed different microbial general structure between them.The Shannon and Chao indexes significantly decreased in the antibiotic treatment group compared to other groups at 7th,14th,and 30th post-infection days,while pretreatment of C.butyricum could maintain the indexes within normal range.At day 7 after infection,the structure of microbiota headed towards normality after the C.butyricum treatment.Conclusions:Early use of C.butyricum treatment could prevent the visceral hypersensitivity in mice induced by C.rodentium infection plus chronic stress.It could also remodel and prevent the microbiota change caused by the infection and chronic stress.It may be a more effective treatment strategy for PI-IBS than antibiotics.