SESSION TITLE: Mechanisms of Infections
SESSION TYPE: Original Investigation Slide
PRESENTED ON: Sunday, April 17, 2016 at 04:00 PM - 05:00 PM
PURPOSE: To investigate the role of interferon regulatory factor-1 (IRF-1) on lipopolysaccharide (LPS) induced reactive oxygen species (ROS) production and mitochondrial damage in macrophages
METHODS:In vitro, stable overexpress or knockdown IRF-1 in murine macrophages RAW264.7 cells was by lentivirus expression vector transfection or lentivirus-mediated RNA interference (RNAi) transfection. The cells were stimulated with LPS (500ng/ml) for 16 hours and were for further study. In vivo, C57BL/6 wild type mice and IRF-1 knockout (KO) mice were randomly given intraperitoneal (IP) injections of PBS or LPS and were sacrificed 16 hours later. Peritoneal macrophages were isolated for further study. The mitochondrial membrane potential (△Ψm) was determined with JC-1; reactive oxygen species (ROS) was measured by DCFH-DA; intracellular ATP, SOD and MDA content were assayed by a commercial kit; Mitochondrial structure damage was also observed by transmission electron microscopy.