title: Liposomal delivery of purified inhibitory-kappaBalpha inhibits tumor necrosis factor-alpha-induced human vascular smooth muscle proliferation.
abstract: Vessel injury results in the elaboration of various cytokines, including tumor necrosis factor-alpha (TNF-alpha), which may influence vascular smooth muscle cell (VSMC) function and contribute to atherogenesis. We tested the hypothesis that TNF-alpha-induced VSMC proliferation requires activation of the transcription factor nuclear factor-kappaB (NF-kappaB), which could be prevented by delivery of the NF-kappaB inhibitory peptide, IkappaBalpha. TNF-alpha induced concentration-dependent human VSMC proliferation, and neutralizing antibody to interleukin-6 reduced TNF-alpha-induced VSMC proliferation by 65%. In TNF-alpha-stimulated VSMCs, there was a 3-fold increase in NF-kappaB-dependent luciferase reporter activity that was associated with degradation of IkappaBalpha. To determine an essential role for NF-kappaB in TNF-alpha-induced VSMC proliferation, recombinant IkappaBalpha was introduced into VSMCs via liposomal delivery. Under these conditions, TNF-alpha-induced NF-kappaB nuclear translocation and DNA binding were inhibited, NF-kappaB-dependent luciferase activity was reduced by 50%, there was no degradation of native IkappaBalpha detected, interleukin-6 production was reduced by 54%, and VSMC proliferation was decreased by 60%. In conclusion, the mitogenic effect of TNF-alpha on human arterial VSMCs is dependent on NF-kappaB activation and may be prevented by exogenously delivered IkappaBalpha. Furthermore, liposomal delivery of endogenous inhibitory proteins may represent a novel, therapeutically accessible method for selective transcriptional suppression in the response to vascular injury.
authors.affiliation: Department of Surgery, University of Colorado Health Sciences Center, Denver, Colo. 80262, USA. craig.selzman@uchsc.edu
authors.forename: C H
authors.initials: CH
authors.lastname: Selzman
authors.forename: B D
authors.initials: BD
authors.lastname: Shames
authors.forename: L L
authors.initials: LL
authors.lastname: Reznikov
authors.forename: S A
authors.initials: SA
authors.lastname: Miller
authors.forename: X
authors.initials: X
authors.lastname: Meng
authors.forename: H A
authors.initials: HA
authors.lastname: Barton
authors.forename: A
authors.initials: A
authors.lastname: Werman
authors.forename: A H
authors.initials: AH
authors.lastname: Harken
authors.forename: C A
authors.initials: CA
authors.lastname: Dinarello
authors.forename: A
authors.initials: A
authors.lastname: Banerjee
mesh_terms: D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000008:administration & dosage* / Q000378:metabolism; D004337:Drug Carriers; D006801:Humans; D020672:I-kappa B Proteins; D015850:Interleukin-6 / Q000502:physiology; D008081:Liposomes; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; D004337:Drug Carriers; D020672:I-kappa B Proteins; D015850:Interleukin-6; D008081:Liposomes; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha