MiR-146a Participates In The Pathogenesis Of Inflammatory Bowel Disease Via Regulating A3AR

Objective: Inflammatory bowel disease(IBD)is a kind of immune intestinal disease with complicated pathogenesis.There is a lack of clinical radical treatment,and the incidence is increasing year by year,which seriously affects the quality of life,and gradually becomes a research hotspot.In recent years,Micro RNA(Mi RNA)have been found to be closely related to IBD,and can affect the pathogenesis,diagnosis and treatment of IBD.It has been reported that miR-146 a is up-regulated in IBD intestinal mucosal tissues,so here we mainly study the expression level of miR-146 a in inflammatory bowel disease and explore its mechanism of regulating A3 AR ’ s involvement in inflammatory bowel disease,so as to provide new ideas and theoretical basis for clarifying the pathogenesis of inflammatory bowel disease.Methods: The intestinal mucosa tissues of 53 patients with inflammatory bowel disease were selected as the IBD group,and 53 healthy intestinal mucosa were selected as the healthy control group.RT-q PCR was used to detect the expression level of miR-146 a in the IBD group and the healthy control group.Using LPS to induce HIEC-6 cells to construct an inflammatory cell model(LPS group),RT-q PCR was used to detect the expression level of miR-146 a in the LPS group.NC inhibitor and miR-146 a inhibitor were transfected into inflammatory cell models,and a blank control group was set up.They were divided into LPS+NC inhibitor group,LPS+miR-146 a inhibitor group,and Control group.The expression levels of miR-146 a in the above three groups were detected by RT-q PCR;Elisa experiment detected pro-inflammatory factor TNF in four groups: LPS,LPS+NC inhibitor,LPS+miR-146 a inhibitor,and Control-α、 IL-1 β and the secretion of IL-6;RT-q PCR and Western blot experiments were used to detect the expression of A3 AR m RNA and protein in four groups: LPS,LPS+NC inhibitor,LPS+miR-146 a inhibitor,and Control.The A3 AR 3 ’-UTR wild-type and mutant double luciferase reporter gene vectors were constructed and co transfected with miR-146 a inhibitor or NC inhibitor into inflammatory cell models,respectively,to observe the changes of Luciferase activity in each group.Results: Compared with the healthy control group,the expression of miR-146 a was significantly upregulated in the IBD group,with a statistically significant difference(P<0.001);Compared with the Control group,the relative expression of miR-146 a significantly increased in the LPS group(P<0.05).Compared with blank control group and NC inhibitor group,the down-regulation of miR-146 a expression decreased the expressions of pro-inflammatory cytokines TNF-α,IL-1β and IL-6,with significant differences(P<0.001).Compared with blank control group,LPS group and LPS+NC inhibitor group,the expression levels of A3 AR m RNA and protein in LPS+miR-146 a inhibitor group were significantly increased,with significant differences(P<0.001).Compared with NC inhibitor combined with A3AR3 ’-UTR wild-type vector co-transfected cells,luciferase activity of miR-146 a inhibitor combined with A3 AR 3’-UTR wild-type vector co-transfected cells increased,the difference was statistically significant(P<0.001).While NC inhibitor and miR-146 a inhibitor were co-transfected with A3 AR 3 ’-UTR mutant vector respectively,the luciferase activity was basically unchanged between the two groups(P>0.05).Conclusion: The increased expression of miR-146 a in intestinal mucosal tissue of inflammatory bowel disease can target the regulation of A3 AR and promote the secretion of Proinflammatory factor TNF-α、IL-1 β and IL-6,is involved in the pathogenesis of inflammatory bowel disease.