title: Milk fat globule epidermal growth factor-factor 8 mitigates inflammation and tissue injury after hemorrhagic shock in experimental animals.
abstract: BACKGROUND
  Insufficient clearance of apoptotic cells leads to increased inflammation and exaggerated organ injury. The opsonizing protein, milk fat globule epidermal growth factor-factor 8 (MFG-E8), upregulates apoptotic cell clearance. The purpose of this study was to determine the degree of apoptotic cell clearance, and whether inflammation, organ injury, and survival are improved after treatment with recombinant human MFG-E8 (rhMFG-E8) after hemorrhagic shock.
  
  
  METHODS
  Male mice underwent a pressure-controlled (25 mm Hg ± 5 mm Hg) model of hemorrhagic shock for 90 minutes. They were resuscitated with normal saline with or without recombinant human MFG-E8 (rhMFG-E8) over 30 minutes. At 3.5-hour postresuscitation, blood and tissue were collected. MFG-E8 levels in the plasma, lungs, and spleen were measured. Apoptotic cell clearance was measured by cleaved caspase-3 levels and TUNEL staining. Neutrophil infiltration was assessed using myeloperoxidase activity in the lungs and spleen. Plasma and tissue levels of proinflammatory cytokines (IL-1β, IL-6, and TNF-α) were measured by ELISA. Finally, a seven-day survival study was also conducted.
  
  
  RESULTS
  MFG-E8 levels in the plasma, lungs, and spleen significantly decreased by 33%, 44%, and 55%, respectively, at 3.5 hour after hemorrhage and resuscitation with rhMFG-E8. Treatment with rhMFG-E8 significantly improved apoptosis, by reducing TUNEL+ cells after treatment and restoring cleaved caspase-3 expression back to baseline. Neutrophil infiltration was blunted by 29% and 41% in the lungs and spleen, respectively. Cytokine expression was also reduced significantly, by 64% to 73% in plasma, 24% to 58% in the lungs, and 49% to 76% in the spleen. Finally, animals demonstrated a superior survival rate over 7 days after treatment with rhMFG-E8.
  
  
  CONCLUSIONS
  The administration of rhMFG-E8 is a potent treatment in animals after hemorrhagic shock.
authors.affiliation: Department of Surgery, Hofstra North Shore-LIJ School of Medicine, Manhasset, New York 11030, USA.
authors.forename: Fangming
authors.initials: F
authors.lastname: Zhang
authors.forename: Kavin G
authors.initials: KG
authors.lastname: Shah
authors.forename: Lei
authors.initials: L
authors.lastname: Qi
authors.forename: Rongqian
authors.initials: R
authors.lastname: Wu
authors.forename: Rafael
authors.initials: R
authors.lastname: Barrera
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Nicastro
authors.forename: Gene F
authors.initials: GF
authors.lastname: Coppa
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000097:blood; D007249:Inflammation / Q000188:drug therapy* / Q000209:etiology; D053583:Interleukin-1beta / Q000097:blood; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008894:Milk Proteins / Q000627:therapeutic use*; D020556:Neutrophil Infiltration / Q000187:drug effects; D012771:Shock, Hemorrhagic / Q000150:complications / Q000188:drug therapy*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D000954:Antigens, Surface; D053583:Interleukin-1beta; C098942:MFGE8 protein, human; D008894:Milk Proteins; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3