Anti-inflammatory effect of Resolvin D1 on LPS-treated MG-63 cells.

Abstract

Inflammation reaction plays an important role in the pathogenesis of ankle fracture. The aim of the present study was to investigate the effect of RvD1 on the inflammatory response and underlying molecular mechanisms in MG-63 cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and ELISA were used to determine the mRNA and protein expression of cytokines extracted from peripheral blood in children with or without ankle fracture, respectively. MG-63 cells were pre-treated with/without RvD1 and stimulated with 1 µg/ml LPS. The cell viability was detected by MTT assay. The production of cytokines from MG-63 cells was assessed by RT-qPCR and western blot, respectively. The expression of p-p38, NF-κB (p50) and cyclooxygenase-2 (COX-2) mRNA and protein were detected by western blot and/or RT-qPCR. The levels of NLRP3, associated recruitment domain (ASC), cleaved caspase1, caspase-1 were measured by RT-qPCR and/or western blot. The levels of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF-α) mRNA and protein were up-regulated in children with ankle fracture compared with healthy children. RvD1 treatment did not induce cytotoxicity in MG-63 cells, but it can inhibit LPS induced MG-63 cell proliferation inhibition. RvD1 was able to dose-dependently reverse LPS induced up-regulation of TNF-α, IL-1β, IL-6 mRNA and protein expression. Furthermore, the LPS induced up-regulation of p-p38, NF-κB (p50), and NLRP3, ASC, cleaved caspase-1/caspase-1, and COX-2 was dose-dependently reversed by RvD1. In conclusion, The present study demonstrated that RvD1 inhibited inflammation though inhibiting MAPKp38/NF-κB pathway and NLRP3 inflammasome expression in MG-63 cells, indicating that it may be an effective drug for the treatment of ankle fracture.