title: [Effects of recombinant human elafin gene transfection on lipopolysaccharide-induced airway mucous hypersecretion].
abstract: OBJECTIVE
  To explore the effects of recombinant human Elafin gene transfection on lipopolysaccharide (LPS)-induced airway mucous hypersecretion.
  
  
  METHODS
  An eukaryotic expression vector pEGFP-C1-Elafin was first constructed and then transfected into NCI-H292 cell. After co-incubation with polymorphonuclear granulocyte (PMN) plus LPS stimulation for 24 hour, the vital force unit of neutrophil elastase (NE) and the content of mucin (MUC) protein in culture media and the level of MUC 5AC mRNA in culture cells were detected with substrate detection technique, enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) respectively.
  
  
  RESULTS
  All parameters in control group showed a low level (0.13 ± 0.03, 0.28 ± 0.06, 0.29 ± 0.04 respectively). And transfecting NCI-H292 cell with pEGFP-C1 at different levels of LPS, all parameters were obviously different (all P < 0.01) and they all showed a dose-dependent relationship. After LPS stimulation (0.5, 5 mg/L), as compared with the transfection of NCI-H292 cell with pEGFP-C1-Elafin, the vital force unit of NE and the content of MUC5AC protein in culture media and the level of MUC 5AC mRNA in transfecting pEGFP-C1 NCI-H292 cell decreased obviously (all P < 0.05). But after LPS stimulation (25 mg/L), as compared with the transfection of NCI-H292 cell with pEGFP-C1-Elafin, no parameter of transfecting pEGFP-C1 NCI-H292 cell had any obvious alteration (0.67 ± 0.06, 0.64 ± 0.08, 0.67 ± 0.07 respectively, all P > 0.05).
  
  
  CONCLUSIONS
  The transfection of recombinant human elafin gene into NCI-H292 cell may inhibit the force of inflammatory telo-effective factor NE induced by a certain level of LPS and airway mucous hypersecretion. But it can not inhibit the effects induced by a high level of LPS.
authors.affiliation: Department of Respiratory Medicine, Zhucheng People's Hospital, Zhucheng 262200, China. xxy121200@163.com
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Xu
authors.forename: Dong-Ming
authors.initials: DM
authors.lastname: Yuan
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Zhou
mesh_terms: D002460:Cell Line; D053492:Elafin / Q000235:genetics*; D005822:Genetic Vectors / Q000187:drug effects; D006801:Humans; D007249:Inflammation; D019272:Leukocyte Elastase / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D055271:Mucin 5AC / Q000032:analysis; D009077:Mucins / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D053492:Elafin; D008070:Lipopolysaccharides; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; D011994:Recombinant Proteins; D019272:Leukocyte Elastase