MiR-155-mediated Deregulation Of GPER1 Plays An Important Role In The Gender Differences Related To Inflammatory Bowel Disease

Inflammatory bowel disease(IBD)is a chronic inflammatory intestinal disease.In recent years,the incidence rate in the world has been increasing year by year,which is a serious threat to human health.Due to the prolonged course of the disease and easy recurrence,there are currently no effective drugs and other factors,which have a huge impact on the emotional,economic,and social conditions of patients and their families.It has become an increasingly serious health problem in the world.IBD-related gender differences,psychological,genetic,environmental,and other factors may be related to gender differences,among which estrogen is considered to be the most important cause.Previous studies have reported that the role of estrogen and nuclear estrogen receptors in gastrointestinal diseases,such as the regulation of estrogen receptors?and?,but the role of G protein-coupled estrogen receptor 1(GPER1)was still poorly understood.GPER1 modulated the immune response,not only affecting intestinal cells,but also immune cells,and has been proven to regulate inflammatory bowel disease and even colon cancer.MiRNAs are involved in the pathogenesis of IBD.Micro RNA-155 is up-regulated in a variety of inflammatory diseases including IBD.It is considered to be an important regulator of T cell response,which is closely related to inflammation and tumorigenesis.Therefore,this topic focuses on the study of gender differences in IBD and related factors,whether miR-155regulates IBD related gender differences through estrogen and its possible mechanisms.This project firstly conducted a meta-analysis of the literature related to gender differences in IBD,and then analyzed IBD patients and control groups to explore the relevant data and influencing factors.Using q PCR,Western blot,IHC and other methods to study the levels of inflammatory factors,estrogen and estrogen receptors in the serum and colon tissues of the two groups of patients.We detected the expression of cytokines AKT1 and NF-?Bp65.GPER1 was predicted to be the target of miR155 by the database.Meanwhile,we detected the expression of miR155.Finally,The weight,survival rate,stool traits and occult blood,survival rate,disease activity index(DAI),colonmucosa damage index(CMDI),colon length,histological grading of the DSS miR155^-/-mice were recorded.We explored the possibility of miR155 regulating GPER1 participates in gender differences in IBD,providing a research basis for miR155 as a noninvasive diagnostic indicator for IBD in the future.Material and Methods:This study was divided into four parts.The first part took IBD patients as the research object.Meta-analysis was used to study the related literature on gender differences in IBD,and analyzed the related issues and influencing factors of gender differences in IBD.In the second part,74 participants(including 50 IBD patients and 24 healthy controls)were enrolled.The content of the study included baseline characteristics,clinical data collection and serological testing.The third part included volunteers who underwent colonoscopy and biopsy,and serological specimens were collected to detect the level of inflammatory factors in serum and colon tissue,the level of estrogen in serum,the protein of estrogen receptor and the downstream signaling pathway of estrogen in tissues,and expression of miR155.The fourth part of the study MiR-155^-/-mice were induced to construct a DSS colitis model.The weight,survival rate,stool traits and occult blood,survival rate,DAI,CMDI,colon length and histological grading of each group of mice were recorded.1.Meta-analysis of literatures were carried out on Pubmed database,Embase database,Medline database,Science Direct database,Cochrane Controlled Test Center database.2.From July 2018 to July 2019,in the Department of Gastroenterology at Daping Hospital,Army Military Medical University,a total of 50 patients with IBD were included in this study,and 24 healthy examinees were randomly selected as the control group.IBD group surveyed diagnose age,course of disease,complications,extraintestinal manifestations,surgery,hospitalizations,smoking history,drinking history,exercise history,drinking history,family history,education level,obesity,marriage,medication,etc.Serological data such as blood routine,biochemistry,and erythrocyte sedimentation rate were collected in each group.3.The serum levels of IL-6,IL-10,TNF?and estrogen in each group were detected.The levels of IL-6,IL-10,TNF?and estrogen receptor in the tissues were detected.The expression of AKT1 and NF-?Bp65 were examined,the database was used to predict the estrogen receptor GPER1 as the target of miR155,and the expression of miR155 was measured by q PCR.4.MiR-155^-/-mice were used to construct a DSS model.The mice were divided into 4groups.The weight,stool blood,survival rate,DAI,CMDI,colon length and histological grading were recorded.Results:1.A total of 7 studies were included in the meta-analysis,including 6299 males and 6987females.Male accounted for 46.4%and female accounted for 53.6%.The results suggested that male smokers had a lower risk of IBD;Male patients had fewer extraintestinal manifestations,especially CD;Male patients used immunosuppressive agents more frequently than female patients;Montreal classification found that male was more severe and female was mild.2.The erythrocyte sedimentation(ESR)level of male with IBD was significantly lower than that of female(P=0.005),and the PLT and ESR of male with CD were lower than those of female(P<0.05).Male CD patients had higher percentage than women in terms of complications and the use of biological agents.(p<0.05).3.The expression of the inflammatory cytokines IL-6 and TNF?was higher in both the serum and tissue of the patients with IBD compared to the control group(p<0.01)and the level increased more significantly in male(p<0.01).The level of estradiol decreased significantly in IBD patients(p<0.05),its expression was significantly decreased in male patients compared with female patients(p<0.001).4.The expression of AKT1 and NF-?Bp65 was significantly higher in the IBD patients than that in the control group(p<0.0001)and the level increased more significantly in male(p<0.0001).The expression of GPER1 in IBD patients was significantly lower than that in the control group(P<0.0001),and the expression of GPER1 in male was significantly lower than that in women(P<0.0001).5.GPER1 is the target of miR-155.The expression of miR-155 was higher in IBD patients((p<0.0001)),and the expression in male was higher than that in female(p<0.001).6.DSS-induced colitis model showed loose stools,bloody stools,weight loss,and varying degrees of shortening of the colon.Pathology showed loss of mucosa,thinning of mucosal thickness,destruction of epithelial structure,incomplete glands,inflammatory cell infiltration.The tissue structure was sparse,and the crypts were destroyed.7.In the DSS induced colitis model of MiR155^-/-mice,the degree of weight loss,survival rate,bloody stool rate,DAI,CMDI,colon length,and histological grading were more mild than those in the DSS WT mice.8.There were gender differences in miR155^-/-mice and the WT mice in the DSS group.The weight loss,survival rate,bloody stool rate,DAI,CMDI,colon length,and histological grading of male mice were more severe than those of the females.Conclusion:1.Meta-analysis showed that there were gender differences in IBD.2.The case-control studies suggested that there were gender differences in IBD patients.3.The inflammation of male IBD patients were more severe than female patients.GPER1participated in the downstream signaling pathway of estrogen.GPER1 exerted a negative regulatory effect on inflammation and participated in the mechanism of gender differences in IBD patients.4.MiR-155 could promote intestinal inflammation.Knocking out miR155 could reduce inflammation.In the pathogenesis of IBD,MiR-155 regulated intestinal inflammation through GPER1.