Nuclear factor κB in LPS stimulated rat alveolar macrophages promotes TNF-α secretion
Qian Gui-sheng
Abstract
Qian Gui-sheng
Abstract
AIM:To investigate the activation of nuclear factor κB(NF-κB) induced by lipopolysaccharides(LPS) in rat alveolar macrophages(AMs) and its regulatory role in tumor necrosis factor(TNF-α) secretion.METHODS:The dynamic activity changes of NF-κB induced by LPS were determined with electrophoretic mobility shift assay(EMSA).Antisense oligonucleotides of NF-κB subunit(p65) were transfected into AMs prior to LPS stimulation.The effect of antisense oligonucleotide transfection on expressions of p65 and TNF-α in supernatant were measured with Western blotting and enzyme linked immunosorbent assay(ELISA),respectively.RESULTS:NF-κB activity increased markedly and reached its peak level at 4 h after LPS stimulation.After transfected with antisense oligonucleotides of NF-κB subunit(p65),expression of p65 and TNF-α in supernatant decreased markedly.CONCLUSION:NF-κB activity has a positive effect on regulating secretion of TNF-α in AMs induced by LPS.
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AIM:To investigate the activation of nuclear factor κB(NF-κB) induced by lipopolysaccharides(LPS) in rat alveolar macrophages(AMs) and its regulatory role in tumor necrosis factor(TNF-α) secretion.METHODS:The dynamic activity changes of NF-κB induced by LPS were determined with electrophoretic mobility shift assay(EMSA).Antisense oligonucleotides of NF-κB subunit(p65) were transfected into AMs prior to LPS stimulation.The effect of antisense oligonucleotide transfection on expressions of p65 and TNF-α in supernatant were measured with Western blotting and enzyme linked immunosorbent assay(ELISA),respectively.RESULTS:NF-κB activity increased markedly and reached its peak level at 4 h after LPS stimulation.After transfected with antisense oligonucleotides of NF-κB subunit(p65),expression of p65 and TNF-α in supernatant decreased markedly.CONCLUSION:NF-κB activity has a positive effect on regulating secretion of TNF-α in AMs induced by LPS.
Key concepts: Tumor necrosis factor alpha, Transfection, Electrophoretic mobility shift assay, Secretion, Molecular biology, NF-κB, Lipopolysaccharide, Oligonucleotide