Reversal of LPS induced endothelial cell TNF synthesis and increased permeability with microencapsulated antisense oligomers to NF-kappaB.
Endothelial cells form the barrier between the circulation and interstitial space. Changes in permeability of endothelial cells allow penetration of inflammatory cells such as polymorphonuclear cells and macrophages to respond to infections and other inflammatory stimuli. Endothelial cells have also been shown to be phagocytic and produce pro-inflammatory cytokines such as TNF. It is the purpose of this study to evaluate endothelial cell phagocytosis of albumin microspheres containing antisense oligonucluetide to NF-kappaB (MASO), the effect of MASO on TNF synthesis after LPS stimulation and the effect of TNF inhibition on the permeability of endothelial cells in vitro. Results were (1) endothelial cells avidly phagocytozed albumin miocrospheres 1.0 and 1.7 microm in size, (2) phagocytosis of microspheres was potentiated by LPS, (3) TNF is synthesized by endothelial cells in cell culture with the peak concentrations occurring 4 h after stimulation with LPS, (4) MASO results in high intracellular concentration of oligomer, (5) MASO inhibits TNF synthesis to a greater extent than equivalent amounts of NF-kappaB antisense in solution and (6) the inhibition of TNF by MASO significantly decreases the permeability of albumin through endothelial cells in vitro.
Zhaowei
Jin
J
Mercer College of Pharmacy and Health Sciences, Atlanta, GA, USA.
D'Souza
Martin J
MJ
Oettinger
Carl W
CW
D002214:Capsules; D002478:Cells, Cultured; D016373:DNA, Antisense; D004337:Drug Carriers; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D004731:Endotoxins / Q000633:toxicity; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D016328:NF-kappa B / Q000378:metabolism*; D009841:Oligonucleotides; D010316:Particle Size; D010539:Permeability / Q000187:drug effects; D048069:Tumor Necrosis Factors / Q000096:biosynthesis*
D002214:Capsules; D016373:DNA, Antisense; D004337:Drug Carriers; D004731:Endotoxins; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009841:Oligonucleotides; D048069:Tumor Necrosis Factors