丁酸调控EV71感染相关肠上皮屏障功能的机制研究OA

Objective:To investigate the regulatory effect of butyrate on intestinal epithelial barrier function during enterovirus A71(EV71)infection and to explore the underlying molecular mechanisms.Methods:An in vitro intestinal epithelial barrier model was established using a Caco-2/HT29-MTX co-culture in Transwell inserts,and an EV71 infection system was subsequently developed.The cells were divided into a control group,an EV71 group,an EV71+butyrate group,an ANT(GPR41/43 antagonist)+EV71 group,and an ANT+EV71+butyrate group.Western blot was used to detect the protein expression of GPR41,GPR43,and the tight junction proteins Occludin,Claudin-1,and ZO-1.The secretion level of mucin 2(MUC2)was measured by ELISA.RT-qPCR was used to measure EV71 RNA levels in the lower chamber of the Transwell system.Results:Compared with the control group,the EV71 group showed decreased expression levels of GPR41,GPR43,and the tight junction proteins Occludin,Claudin-1,and ZO-1(P＜0.05),along with a marked reduction in MUC2 secretion(P＜0.05).Compared with the EV71 group,the EV71+butyrate group exhibited increased expression of the above proteins and elevated MUC2 secretion levels(P＜0.05).Compared with the EV71+butyrate group,the ANT+EV71+butyrate group exhibited lower expression of GPR41,GPR43,Occludin,Claudin-1,and ZO-1,together with reduced MUC2 secretion(P＜0.05).RT-qPCR results showed that EV71 RNA expression was lower in the EV71+butyrate group than in the EV71 group(P＜0.05),whereas it was higher in the ANT+EV71+butyrate group than in the EV71+butyrate group(P＜0.05).Conclusion:Butyrate may enhance intestinal epithelial barrier function and reduce EV71 transepithelial passage through GPR41/43-related signaling,possibly by upregulating tight junction protein expression and promoting MUC2 secretion.