Circ-NCX1 inhibits LPS-induced chondrocyte apoptosis by regulating the miR-133a/SIRT1 axis.

Abstract

Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ-NCX1 in lipopolysaccharide (LPS)-induced injury in SW1353 chondrocytes, an in vitro model of OA. The levels of circ-NCX1, miR-133a, and related apoptotic proteins were determined by RT-qPCR. MTT assay was used to evaluate the cell viability. The apoptosis was determined by flow cytometry, whereas the expression of apoptosis proteins was detected by Western blot. Immunofluorescence was used to detect cleaved caspase-3 expression in cells. Luciferase reporter assay was used to verify the interaction between circ-NCX1 and miR-133a, and between miR-133a and Silent information regulator 2 homolog 1 (Sirt1). The results showed that the overexpression of circ-NCX1 significantly upregulated the chondrocyte viability and proliferation, and alleviated apoptosis in LPS-induced SW1353 cells. Immunofluorescence results showed that the overexpression of circ-NCX1 significantly reduced expression of LPS-stimulated cleaved Caspase-3. The RT-qPCR results showed that the overexpression of circ-NCX1 inhibited mRNA levels of cleaved Caspase-3 and Bax, and promoted mRNA levels of Bcl-2. Luciferase reporter assay showed that circ-NCX1 targeted miR-133a, and miR-133a directly targeted the Sirt1. In addition, overexpression of circ-NCX1 inhibited chondrocyte apoptosis and promoted Akt phosphorylation via the miR-133a/Sirt1 axis in LPS-induced chondrocytes. In conclusion, circ-NCX1 may serve as an important regulator of LPS-induced chondrocyte apoptosis through the miR-133a/Sirt1 axis, and may be involved in the development of OA.