The Aryl Hydrocarbon Receptor Are Involved In The Formation Of Inflammation By Mediated Abnormal Differentiation Of Th17/Treg In Crohn's Disease

The aryl hydrocarbon receptor are involved in the formation of inflammation by mediated abnormal differentiation of Th17/Treg in Crohn's diseaseBackgroundCrohn's disease(CD),one of major phenotype of inflammatory bowel disease(IBD),is characterized by chronic relapsing inflammatory disorder of the gastrointestinal tract.The etiology of CD is complex,which is genetic,environmental,microorganism,immune and other factors involved in its development,especially the immune factor occupies an important position.Concurrently,Th17/Treg imbalance is a hot research topic in recent years.Domestic and foreign studies have shown that regulatory T cells(regulatory T cell,Treg)are a subgroup of T cells with immunosuppressive effect,which inhibit T cell proliferation and activation.And Treg cells also ameliorates intestinal inflammation of IBD patients.The role of Thl7 cells in IBD remains controversial.Studies have shown that Th17 cells exacerbated intestinal inflammation was partially dependent on cytokines expression.On the contrary,some studies suggested that Th17 had certain protective cells in intestinal mucosa.Aryl hydrocarbon receptor(AhR),a ligand-dependent transcriptional factor,is widely expressed by various immune and non-immune cells.AhR has been shown to regulate the differentiation of Th17 and Treg cells.And studies have confirmed that AhR protein expression in patients with CD was downregulated.Lacking of AhR can lead to forkhead box P3(Foxp3)expression downregulation and upregulation of retinoic acid receptor-related orphan nuclear receptor gamma(RORyt).Foxp3,RORyt are respectively important transcription factors for the differentiation of Treg and Th17 cells.The present study investigated whether the AhR are involved in the formation of inflammation by mediated abnormal differentiation of Thl7/Treg in patients with Crohn's disease through regulation of Foxp3,RORyt protein expressions.AimTo investigate whether AhR are involved in the formation of inflammation by mediated abnormal differentiation of Th17/Treg in patients with Crohn's disease through detect the expressions of AhR,Foxp3,RORyt in colon of active CD patients and the alterations in frequency of Treg and Th17 cells in peripheral blood and analyse the relationships between the proportion of Treg cells and serum inflammatory biomarkers with disease activity of CD patients as well as clinical significance.Methods1.Forty-one patients with CD(27 quiescent and 14 active CD patients)were enrolled in our study.Twelve cases who had their gastrointestinal polyps treated and will have gastrointestinal endoscopy to check as normal controls.All CD patients and normal controls were from Department of Gastroenterology,First Affiliated Hospital of Nanjing Medical University(Nanjing,Jiangsu).2.The disease activity of CD was evaluated by crohn's disease activity index(CDAI),remission:<150,activity:?150,mild:150-220,moderate:221-450,severe:>450.3.The protein of AhR,Foxp3,RORyt was evaluated by western blot.4.The percentage of Treg and Th17 cells in peripheral blood from CD patients and control group were detected by flow cytometry.5.Blood platelet(PLT),mean platelet volume(MPV),platelet distribution width(PDW),erythrocyte sedimentation rate(ESR)and serum C-reactive protein(CRP)level were measured.Results1.In peripheral blood,ESR and serum CRP levels increased and MPV obviously decreased in active CD patients compared to quiescent CD patients.2.Compared with normal controls,the expression of AhR protein in the intestinal tissue was decreased in active CD patients.Meanwhile the RORyt expression was upregulated,and the Foxp3 expression was downregulated.3.In peripheral blood,the propotion of Treg cells in active CD patients was significantly decreased compared with patients in remission and control group.4.Compared with quiescent CD patients,active CD patients had higher percentage of Thl7 cells in peripheral blood.5.Negative correlation was found between proportion of Treg cells and ESR,serum CRP levels as well as CDAI.In addition,there was a positive correlation between the proportion of Treg cells and MPV.ConclusionAhR might be involved in the formation of inflammation by mediated abnormal differentiation of Th17/Treg cells in patients with Crohn's disease through regulation of Foxp3,RORyt protein expressions.The aryl hydrocarbon receptor are involved in the formation of inflammation by mediated abnormal differentiation of intestinal ILC3/ILC1 in Crohn's diseaseBackground The abnormal differentiation of intestinal innate lymphoid cells ILC3 and ILC1 exist in autoimmune disease.ILC3 decreased and ILC1 increased in Crohn's disease(CD)patients,suggesting that CD patients have abnormal intestinal ILC3/ILC1 alteration,but the mechanism of abnormal alteration of ILC3/ILC1 keep unclear.The present study investigated the aberrant colonic mucosal ILC3/ILC1 in active CD patients and 2,4,6-trinitrobenzene sulphonic acid(TNBS)-induced colitis mice.Aim To investigate whether the aryl hydrocarbon receptor(AhR)are involved in the formation of inflammation by mediated abnormal differentiation of intestinal ILC3/ILC1 in patients with Crohn's disease through detect the expressions of aryl hydrocarbon receptor(AhR)in colon of active CD patients and normal controls by western blot and the ILC3/ILC1 were investigated in CD patients,normal controls and TNBS-induced colitis mice(AhR+/+,AhR*;*)by immunofluorescence.Methods1.Active CD patients and normal controls were recruited.The distribution ofILC3/ILC1 cells in colonic mucosa and the lamina propria were observed by immunofluorescence staining.2.The protein of AhR was evaluated by western blot.3.The distribution of ILC3/ILC1 were investigated in TNBS-induced colitis mice(AhR+/+,AhR-/-)by inrnumofluorescence.Results1.Compared with normal controls,the number of ILC1 cells was mainly increased in colonic mucosa and the lamina propria and the number of ILC3 cells was mainly decreased in colonic mucosa of patients with active CD.2.Compared with normal controls,the expression of aryl hydrocarbon receptor(AhR)in the intestinal tissue was decreased in active CD patients.3.The intestinal inflammation in TNBS-induced colitis AhR-A mice(AhRv-/-)was more severe than wild type mice(AhR+/+).4.The number of ILC1 in TNBS-induced colitis AhR-/- mice was increased while ILC3 decreased.Conclusion These findings suggest that AhR may mediate abnormal differentiation of ILC3/ILC1,promotes the pathogenesis of CD and play an important role in disease activity and mucosal damage.