Down-Regulation of MCT1 Ameliorates LPS-Induced Cell Injury in Murine Chondrocyte-like ATDC5 Cells by Regulation of PFKFB3

Abstract

: MCT1 is an important regulator in glycolysis and has significant effects on inflammatory responses and osteoclast differentiation etc. This study was to study the effects and mechanism of MCT1 in chondrocytes injury and inflammatory responses. ATDC5 cells with stably transfection of MCT1shRNA were treated with 5 μg/mL of LPS. Cell viability was de termined by MTT assay. The mRNA and protein expressions were detected by qRT-PCR and western blotting, respectively. The concentrations of cytokines in culture medium were measured by ELISA. ROS generation was tested by 2,7-dichloro fluorescein diacetate (DCFH-DA). The results showed that MCT1 was increased by LPS treatment in ATDC5 cells in a dose dependent manner. MCT1 knockdown improved the survival of LPS-treated ATDC5 cells. MCT1 knockdown also de creased LPS-induced expression of pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNFα) and oxidative stress media tors (iNOS, COX-2 and NOX-4) in ATDC5 cell. Importantly, PFKFB3 overexpression reversed the anti-inflammatory and anti-oxidative stress effects of MCT1 knockdown in LPS-induced ATDC5 cells. These results indicated that MCT1 knock out decreased the expression of inflammatory mediators and oxidative stress mediators induced by LPS through regulating PFKFB3. The study provides a potential target for the prevention or treatment of osteoarthritis (OA) and rheumatoid arthritis (RA).