title: [Ac-hE-18A-NH2 inhibits the inflammatory response induced by ox-LDL via inhibiting NF-κB activation in RAW264.7 macrophages].
abstract: OBJECTIVE
  To evaluate the effect of Ac-hE-18A-NH2 on TNF-α secretion and mRNA expression in ox-LDL-stimulated RAW264.7 macrophages and to elucidate the possible mechanisms.
  
  
  METHODS
  Macrophages were incubated in the medium containing various concentrations of Ac-hE18A-NH2 (1-50 μg/mL) with ox-LDL (50 μg/mL) stimulated. The TNF-α level and intracellular cholesterol content were measured by commercially available quantitation kits following the manufacturer's instructions. TNF-α and ATP-binding cassette transporter A1 (ABCA1) mRNA expression were detected by real-time PCR. ABCA1 and IκB protein -expression in the macrophages were determined by Western blot. NF-κB activity was evaluated by electrophoretic mobility shift assay (EMSA).
  
  
  RESULTS
  Ox-LDL stimulation induced a significant increase in TNF-α secretion, mRNA expression, cholesterol accumulation and nuclear factor-κB (NF-κB) activity in RAW264.7 macrophages. Ac-hE-18A-NH2 reduced TNF-α secretion and mRNA expression, up-regulated the ABCA1 mRNA and protein expression, reduced the intracellular cholesterol content, and inhibited NF- κB activation in a dose-dependent manner. Under the same condition and the same concentration, Ac-hE-18A-NH2 was more efficient than D-4F (apoA-I mimetic peptide) in inhibiting the inflammatory response induced by ox-LDL in the macrophages.
  
  
  CONCLUSIONS
  Ac-hE-18A-NH2 may suppress TNF-α secretion and mRNA expression in ox-LDL stimulated RAW264.7 macrophages via IκB-NF-κB signaling pathway. The anti-inflammatory effect of Ac-hE-18A-NH2 is better than that of apoA-I mimic peptide D-4F.
authors.affiliation: Department of Cardiology, People's Hospital of Hunan, Changsha 410005,China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Xie
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Shuiping
authors.initials: S
authors.lastname: Zhao
mesh_terms: D064286:ATP Binding Cassette Transporter 1 / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D005786:Gene Expression Regulation; D020672:I-kappa B Proteins / Q000378:metabolism; D007249:Inflammation / Q000378:metabolism*; D008074:Lipoproteins / Q000494:pharmacology*; D008077:Lipoproteins, LDL; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology*; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D064286:ATP Binding Cassette Transporter 1; D002784:Cholesterol; D020672:I-kappa B Proteins; D008074:Lipoproteins; D008077:Lipoproteins, LDL; D016328:NF-kappa B; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; C577833:Abca1 protein, mouse; C478448:Ac-hE18A-NH(2); C099252:oxidized low density lipoprotein