LncRNA ALDB-898 modulates intestinal epithelial cell damage caused by Clostridium perfringens type C in piglet by regulating ssc-miR-122-5p/OCLN signaling.

Diarrhea of piglets caused by Clostridium perfringens type C (C. perfringens type C) infection is a global problem afflicting piglet production. Long noncoding RNA (LncRNA) and microRNA (miRNA) have emerged as critical regulators of this pathological process, but the underlying molecular mechanisms remain unclear. In this study, we first observed the expression changes of ALDBSSCG0000000898 (ALDB-898) and ssc-miR-122-5p in infected ileum tissue of piglets with C. perfringens type C, and then used C. perfringens beta2 toxin (CPB2) to induce intestinal porcine epithelial cells (IPEC-J2) to construct an injury model. Cytometry kit 8 (CCK-8), lactate dehydrogenase (LDH), real-time quantitative polymerase chain reaction (RT-qPCR), Western blot, flow cytometry and fluorescein isothiocyanate-dextran 4 (FITC-Dextran 4) flux assays were performed to study the effect of ALDB-898 and ssc-miR-122-5p in apoptosis, inflammation and intestinal barrier damage and inflammatory in IPEC-J2 cells induced by CPB2. In addition, dual-luciferase reporter gene analysis was performed to confirm the relationship between ssc-miR-122-5p and ALDB-898 or ssc-miR-122-5p and occludin (OCLN), respectively. There were lower expression levels of ALDB-898 and OCLN and higher expression levels of ssc-miR-122-5p in diarrhea piglets caused by Clostridium perfringens type C. ALDB-898 and OCLN were significantly decreased and ssc-miR-122-5p was increased in IPEC-J2 after exposure to the CPB2 in a dose- and time-dependent manner. ALDB-898 overexpression mitigated CPB2-induced cell injury by promoting viability, restraining apoptosis, cytotoxicity, and inflammatory response, as well as weakening the destruction of the intestinal barrier. Further mechanisms disclosed that ALDB-898 functioned as a competing endogenous RNA (ceRNA) via binding to ssc-miR-122-5p, and OCLN was a target of ssc-miR-122-5p. Importantly, the ssc-miR-122-5p mimic led to abolishing the protective function of ALDB-898 on CPB2-induced IPEC-J2 cell damage, and the addition of OCLN reversed the negative impact of ssc-miR-122-5p, thereby restoring the protection of ALDB-898. Our data showed that ALDB-898 could enhance the expression of OCLN through competitive binding ssc-miR-122-5p to suppress CPB2-induced damage. The ALDB-898/ssc-miR-122-5p/OCLN signaling may be a candidate therapeutic pathway for diarrhea of piglets.