脓毒症发病机制及其防治新进展解剖.ppt

S1PR2 signaling suppresses phagocytic function of alveolar macrophages (AMs). Part 2. Impact of S1PR2 on the regulation of macrophage phagocytosis Part 2. Conclusion Bone marrow chimeras Alveolar macrophage deletion Phagocytosis and bactericidal assay Deletion of S1PR2 in the BM-derived cells leads to increased bacterial clearance activity in the lung. Deletion of S1PR2 in AMs improves pulmonary bacterial clearance. S1PR2 deficiency enhances AMs phagocytic function. in vivo in vivo ex vivo Contents The relationship between S1PR2 and the outcome of spsis Impact of S1PR2 on the regulation of macrophage phagocytosis The molecular mechanisms of S1PR2 in inhibiting macrophage phagocytosis The relationship between S1PR2 expression in monocytes and the outcome of septic patients GTPase cascades involved in cytoskeleton organization Part 3. Background Ronald S. Flannagan, et al. Annu Rev Pathol. 2012. Giovanna Chimini, et al. Nature cell biology. 2000. Sandrine Etienne-Manneville, et al. Nature. 2002. Actin polymerization is the key step of phagocytosis Rho GTPase cascades involved in actin polymerization In vivo, S1P concentration increased from 0 h to 18 h after lung infection. Exogenous S1P (from 100 nM to 5 ?M) reduced phagocytosis by ~40% in WT BMDMs, but not in S1pr2-/- BMDMs. Part 3. The molecular mechanisms of S1PR2 in inhibiting macrophage phagocytosis A B Part 3. The molecular mechanisms of S1PR2 in inhibiting macrophage phagocytosis Low S1P concentration High S1P concentration: S1pr2+/+ S1pr2-/- BMDM E. coli (30 min or 60 min) Confocal microscopy：F-actin, IQGAP1 distribution； Pull Down：RAC1-GTP； Western blot：F-actin； Co-Immunoprecipitation：IQGAP1-RAC1 binding； Small interfering RNA: IQGAP1 S1pr2+/+ S1pr2-/- BMDM S1P 100nM + E. coli (30 min) Confocal microscopy：F-actin and RAC1distribution； Pull Down：RhoA-GTP Part 3. The molecular mechanisms of S1PR2 in inhibiting macrophage phagocytosis At early stages of E. coli intratracheal infection (lo