Effects of recombinant human elafin gene transfection on lipopolysaccharide-induced airway mucous hypersecretion

Abstract

To explore the effects of recombinant human Elafin gene transfection on lipopolysaccharide (LPS)-induced airway mucous hypersecretion. 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. 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). 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.