Lnc‐ANRIL modulates the immune response associated with NF‐κB pathway in LPS‐stimulated bovine mammary epithelial cells

Abstract

AbstractBackgroundThe antisense noncoding RNA in the INK4 locus (ANRIL) has been confirmed related to multiple disease progression, but the role and exact mechanisms of lnc‐ANRIL in lipopolysaccharide (LPS)‐induced inflammation of bovine mammary epithelial cells (MAC‐T) remain unclear.AimsThis manuscript focused on expounding the functional role of lnc‐ANRIL through experiments performed in MAC‐T.MethodsAt the in vitro level, we established a Bovine mammary epithelial cell (BMEC) cell model of mastitis by LPS treatment. Transfection of siRNA was examined by immunofluorescence localization and RT‐qPCR. CCK8, clonogenic assay and EdU were used to detect the proliferation ability of the cells. Cell cycle and apoptosis were detected by flow cytometry and Western blot. The levels of inflammatory factors and oxidative stress markers were detected by ELISA kits.ResultsCell Counting Kit‐8, colony formation, and 5‐ethynyl‐20‐deoxyuridine were adopted and the data illustrated that LPS could significantly suppress the cell proliferation, while knockdown of lnc‐ANRIL expression obviously promoted MAC‐T cell proliferation compared with LPS or LPS + si‐NC group. Flow cytometry analysis demonstrated that lnc‐ANRIL could induce MAC‐T cell apoptosis. In addition, downregulation of lnc‐ANRIL affected LPS‐induced immune response by regulating inflammatory factor expressions and modulating the nuclear factor kappa B (NF‐κB) axis in MAC‐T cells.ConclusionOur results suggest that lnc‐ANRIL is involved in the regulation of cell proliferation, cell cycle, and cell apoptosis of MAC‐T cells, and plays an important role in the inflammatory and immune response of MAC‐T cells through the regulation of the NF‐κB pathway, proposing new therapeutic strategies for the treatment of innate immune response‐related disease such as bovine mastitis.