ssc-micorRNA-132 suppresses the Clostridium perfringens beta2 toxin induced inflammation and apoptosis of IPEC-J2 cells via targeting DACH1

Clostridium perfringens (C. perfringens) beta2 (CPB2) is the main virulence factor secreted from C. perfringens type C, which caused diarrhea characterized by high mortality in pig, especially newborn piglets. Our previous research found that ssc-miR-132 displayed decreased expression in piglets diarrhea after infected with C. perfringens type C compared with normal piglets. We speculated that ssc-miR-132 may play an important role in the diarrhea. However, the function of ssc-miR-132 in the diarrhea is limited. Thus, we overexpressed and knocked down ssc-miR-132 in intestinal porcine epithelial (IPEC-J2) cells, and then treated the cells with recombinant CPB2 (rCPB2) toxin (20 g/mL). Our results showed that ssc-miR-132 was significantly decreased after treated with rCPB2 toxin. In addition, overexpression of ssc-miR-132 reduced the expression of lactate dehydrogenase (LDH) and tumor necrosis factor (TNF-), interleukin-6 (IL-6) and interleukin-8 (IL-8) caused by rCPB2 toxin. The CCK8, Edu and TUNEL staining showed that overexpression of ssc-miR-132 weakened the inhibition of rCPB2 toxin on cell proliferation and reduced the promotion of cell apoptosis; while inhibition of ssc-miR-132 had opposite results. The dual luciferase experiment showed that dachshund family transcription factor 1 (DACH1) was the target gene of ssc-miR-132. Silencing DACH1 was consistent with the results of overexpression of ssc-miR-132, and reversed the apoptosis and inflammation caused by rCPB2 toxin. Overexpression of DACH1 weakened the role of ssc-miR-132 in rCPB2 toxin -induced inflammation and apoptosis. In summary, ssc-miR-132 inhibited rCPB2 toxin-induced apoptosis and inflammation in IPEC-J2 cells by targeting DACH1.