LAUSR

AIMS ZO-1 is a key regulatory tight junction protein that plays an important role in maintaining gastrointestinal health. In this study, we investigated the protective effect and regulation mechanism of the probiotic Enterococcus faecium HDRsEf1 on tight junction protein ZO-1 at the cellular and molecular levels. METHODS AND RESULTS We established LPS-induced intestinal epithelial cell injury model, and detected the protective effect of HDRsEf1 on ZO-1 in IPEC-J2 cells by Real-time PCR and Western blot. The results showed that HDRsEf1 inhibited the downregulation of ZO-1 expression induced by LPS. HDRsEf1 stabilized the destruction of the ZO-1 structure caused by LPS in an immunofluorescence assay. Through gene overexpression and siRNA interference tests, we found that transcription factor AP-1 inhibited the level of ZO-1 expression. Silencing experiment further supported that the protective effect of HDRSEF1 might mediated by suppression of LPS-provoked activation of ASK1/MKK7/JNK signalling pathways. In addition, HDRsEf1 could stabilize ZO-1 expression by increasing TLR2 expression and competing with LPS for the TLR4 binding site. More interestingly, we also found that HDRsEf1 could stabilize ZO-1 expression through inhibiting the production of TNF-α induced by LPS. CONCLUSIONS HDRsEf1 could protect the IPEC-J2 cell against LPS induced down-regulation of ZO-1 expression by inhibiting the activation of TLR2/4-mediated JNK-AP-1 and signalling cascade and the production of TNF-α. SIGNIFICANCE AND IMPACT OF THE STUDY This study can provide a theoretical basis for probiotics to regulate the expression of intestinal tight junction proteins, and supply technical support for probiotics to prevent and treat animal intestinal infectious diseases.