NF-kappaB activation is inhibited in human pulmonary epithelial cells transfected with alpha-melanocyte-stimulating hormone vector.
alpha-Melanocyte-stimulating hormone (alpha-MSH) modulates inflammation. We investigated the influence of alpha-MSH on NF-kappaB activation in human pulmonary epithelial cells (A549) using a plasmid vector encoding alpha-MSH (pCMV-ssMSH). Electrophoretic mobility shift assays demonstrated that NF-kappaB activation induced by lipopolysaccharide was inhibited in A549 cells transfected with pCMV-ssMSH. Western blot analysis revealed that this inhibition was linked to preservation of expression of IkappaBalpha protein. Chloramphenicol acetyltransferase assay indicated that NF-kappaB-dependent reporter gene expression was suppressed in A549 cells transfected with pCMV-ssMSH. The findings indicate that anti-inflammatory actions are exerted via modulation of NF-kappaB activation by preservation of IkappaBalpha protein in human pulmonary epithelial cells transfected with alpha-MSH vector. We showed a possibility of gene therapy for chronic inflammatory lung diseases.
Ichiyama
T
T
Department of Physiology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75235-9040, USA. ichiyama@po.cc.yamaguchi-u.ac.jp
Okada
K
K
Campbell
I L
IL
Furukawa
S
S
Lipton
J M
JM
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides / Q000494:pharmacology; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D011014:Pneumonia / Q000235:genetics / Q000628:therapy; D021381:Protein Transport / Q000187:drug effects; D014162:Transfection; D000521:alpha-MSH / Q000235:genetics / Q000378:metabolism*
D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D009687:Nuclear Proteins; D000072000:NF-KappaB Inhibitor alpha; D000521:alpha-MSH