Oxidative stress induces NF kappa B DNA binding and inducible NOS mRNA in human epithelial cells.
Free radicals generated by a partial reduction of O2 pose a serious threat to tissues and vital organs and cells. The major site of interaction between the lung and inhaled oxidants is the epithelium. We have examined the effect of pyrogallol, an O2- generator, on the ability of human epithelial cells to produce active DNA binding proteins and inducible nitric oxide synthase (iNOS) mRNA in cultured A549 epithelial cells. NF kappa B binding in the nuclei of these cells was determined by electrophoretic mobility shift assays. iNOS mRNA was measured using reverse transcription of PCR. There was a time- and concentration-dependent induction of NF kappa B binding, followed by a time and dose dependent increase in iNOS mRNA levels. These results suggest that in airways the initial response to oxidative stress may be to induce NF kappa B-responsive genes, such as iNOS, which may play an important role in defending the airway against oxidative stress.
Adcock
I M
IM
Department of Thoracic Medicine, National Heart and Lung Institute, London, U.K.
Brown
C R
CR
Kwon
O
O
Barnes
P J
PJ
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism; D017931:DNA Primers; D004790:Enzyme Induction; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008168:Lung; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D019001:Nitric Oxide Synthase; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011748:Pyrogallol / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D012137:Respiratory System; D013481:Superoxides
D017931:DNA Primers; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011748:Pyrogallol; D013481:Superoxides; D004247:DNA; D019001:Nitric Oxide Synthase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000594:Amino Acid Oxidoreductases