title: Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract.
abstract: OBJECTIVE
  Macrophages play a pivotal role in initiating, maintaining, and resolving host inflammatory/immune responses but may cause recalcitrant inflammation and tissue damage if not controlled. Clinically, amniotic membrane (AM) transplantation suppresses inflammation in ocular surface reconstruction. Experimentally, the authors and others have reported that AM facilitates macrophage apoptosis. However, it remains unclear whether such anti-inflammatory activity is retained in AM extract (AME).
  
  
  METHODS
  Herein the authors demonstrate in resting and activated (by interferon [IFN]-gamma, lipopolysaccharide [LPS], or IFN-gamma/LPS) murine monocyte/macrophage RAW264.7 cells that AME suppresses cell spreading and reduces actin filaments determined by phalloidin staining and Western blotting of Triton X-100 extracted cell lysate.
  
  
  RESULTS
  Western blot and immunocytochemistry staining showed AME downregulates the expression of such cell surface markers as CD80, CD86, and major histocompatibility complex class 2 antigen. Cell growth/viability is inhibited whereas cell apoptosis is enhanced by AME. Accordingly, secreted proinflammatory cytokines such as TNF-alpha and IL-6 are reduced, but anti-inflammatory cytokine IL-10 is upregulated.
  
  
  CONCLUSIONS
  Collectively, these results suggest that, similar to amniotic membrane, AME retains anti-inflammatory activities and does so by downregulating activation and inducing apoptosis in macrophages.
authors.affiliation: TissueTech, Inc and the Ocular Surface Center, Miami, Florida 33173, USA.
authors.forename: Hua
authors.initials: H
authors.lastname: He
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Szu-Yu
authors.initials: SY
authors.lastname: Chen
authors.forename: Shan
authors.initials: S
authors.lastname: Zhang
authors.forename: Ying-Ting
authors.initials: YT
authors.lastname: Chen
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Hayashida
authors.forename: Ying-Ting
authors.initials: YT
authors.lastname: Zhu
authors.forename: Scheffer C G
authors.initials: SC
authors.lastname: Tseng
mesh_terms: D000199:Actins / Q000378:metabolism; D000650:Amnion / Q000166:cytology*; D000818:Animals; D017209:Apoptosis; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D015153:Blotting, Western; D002457:Cell Extracts / Q000494:pharmacology*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D009000:Monocytes / Q000378:metabolism / Q000473:pathology*
chemical_list: D000199:Actins; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D002457:Cell Extracts; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D008070:Lipopolysaccharides; D007371:Interferon-gamma