title: In vitro and in vivo nuclear factor-kappaB inhibitory effects of the cell-penetrating penetratin peptide.
abstract: Penetratin is a cationic cell-penetrating peptide that has been frequently used for the intracellular delivery of polar bioactive compounds. Recent studies have just revealed the major role of polyanionic membrane proteoglycans and cholesterol-enriched lipid rafts in the uptake of the peptide. Both proteoglycans and lipid-rafts influence inflammatory processes by binding a wide array of proinflammatory mediators; thus, we decided to analyze the effect of penetratin on in vitro and in vivo inflammatory responses. Our in vitro luciferase gene assays demonstrated that penetratin decreased transcriptional activity of nuclear factor-kappaB (NF-kappaB) in tumor necrosis factor (TNF)-stimulated L929 fibroblasts and lipopolysaccharide-activated RAW 264.7 macrophages. Penetratin also inhibited TNF-induced intercellular adhesion molecule-1 expression in human endothelial HMEC-1 cells. Exogenous heparan sulfate abolished the in vitro NF-kappaB inhibitory effects of the peptide. Uptake experiments showed that penetratin was internalized by all of the above-mentioned cell lines in vitro and rapidly entered the cells of the lung and pancreas in vivo. In an in vivo rat model of acute pancreatitis, a disease induced by elevated activities of stress-responsive transcription factors like NF-kappaB, pretreatment with only 2 mg/kg penetratin attenuated the severity of pancreatic inflammation by interfering with IkappaB degradation and subsequent nuclear import of NF-kappaB, inhibiting the expression of proinflammatory genes and improving the monitored laboratory and histological parameters of pancreatitis and associated oxidative stress.
authors.affiliation: Department of Medical Chemistry, University of Szeged, Dóm tér 8, H-6720 Szeged, Hungary. tletoha@yahoo.com
authors.forename: Tamás
authors.initials: T
authors.lastname: Letoha
authors.forename: Erzsébet
authors.initials: E
authors.lastname: Kusz
authors.forename: Gábor
authors.initials: G
authors.lastname: Pápai
authors.forename: Annamária
authors.initials: A
authors.lastname: Szabolcs
authors.forename: József
authors.initials: J
authors.lastname: Kaszaki
authors.forename: Ilona
authors.initials: I
authors.lastname: Varga
authors.forename: Tamás
authors.initials: T
authors.lastname: Takács
authors.forename: Botond
authors.initials: B
authors.lastname: Penke
authors.forename: Erno
authors.initials: E
authors.lastname: Duda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D057846:Cell-Penetrating Peptides; D002478:Cells, Cultured; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010179:Pancreas / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D019283:Pancreatitis, Acute Necrotizing / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D016328:NF-kappa B; C414409:penetratin