title: Antiarthritic effect of bee venom: inhibition of inflammation mediator generation by suppression of NF-kappaB through interaction with the p50 subunit.
abstract: OBJECTIVE
  To investigate the molecular mechanisms of the antiarthritic effects of bee venom (BV) and melittin (a major component of BV) in a murine macrophage cell line (Raw 264.7) and in synoviocytes obtained from patients with rheumatoid arthritis.
  
  
  METHODS
  We evaluated the antiarthritic effects of BV in a rat model of carrageenan-induced acute edema in the paw and in a rat model of chronic adjuvant-induced arthritis. The inhibitory effects of BV and melittin on inflammatory gene expression were measured by Western blotting, and the generation of prostaglandin E(2) (PGE(2)) and nitric oxide (NO) and the intracellular calcium level were assayed. NF-kappaB DNA binding and transcriptional activity were determined by gel mobility shift assay or by luciferase assay. Direct binding of BV and melittin to the p50 subunit of NF-kappaB was determined with a surface plasmon resonance analyzer.
  
  
  RESULTS
  BV (0.8 and 1.6 mug/kg) reduced the effects of carrageenan- and adjuvant-induced arthritis. This reducing effect was consistent with the inhibitory effects of BV (0.5, 1, and 5 mug/ml) and melittin (5 and 10 mug/ml) on lipopolysaccharide (LPS; 1 mug/ml)-induced expression of cyclooxygenase 2, cytosolic phospholipase A(2), inducible NO synthase, generation of PGE(2) and NO, and the intracellular calcium level. BV and melittin prevented LPS-induced transcriptional and DNA binding activity of NF-kappaB via the inhibition of IkappaB release and p50 translocation. BV (affinity [K(d)] = 4.6 x 10(-6)M) and melittin (K(d) = 1.2 x 10(-8)M) bound directly to p50.
  
  
  CONCLUSIONS
  Target inactivation of NF-kappaB by directly binding to the p50 subunit is an important mechanism of the antiarthritic effects of BV.
authors.affiliation: College of Pharmacy, Chungbuk National University, 48 Gaesin-dong, Heungduk-gu, Cheongju, Chungbuk 361-763, South Korea.
authors.forename: Hye Ji
authors.initials: HJ
authors.lastname: Park
authors.forename: Seong Ho
authors.initials: SH
authors.lastname: Lee
authors.forename: Dong Ju
authors.initials: DJ
authors.lastname: Son
authors.forename: Ki Wan
authors.initials: KW
authors.lastname: Oh
authors.forename: Ki Hyun
authors.initials: KH
authors.lastname: Kim
authors.forename: Ho Sueb
authors.initials: HS
authors.lastname: Song
authors.forename: Goon Joung
authors.initials: GJ
authors.lastname: Kim
authors.forename: Goo Taeg
authors.initials: GT
authors.lastname: Oh
authors.forename: Do Young
authors.initials: DY
authors.lastname: Yoon
authors.forename: Jin Tae
authors.initials: JT
authors.lastname: Hong
mesh_terms: D000818:Animals; D001168:Arthritis / Q000517:prevention & control*; D001169:Arthritis, Experimental / Q000517:prevention & control; D001172:Arthritis, Rheumatoid / Q000378:metabolism / Q000473:pathology; D001514:Bee Venoms / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002351:Carrageenan; D002460:Cell Line; D015232:Dinoprostone / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004487:Edema / Q000139:chemically induced / Q000378:metabolism; D006801:Humans; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors*; D007425:Intracellular Membranes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008156:Luciferases / Q000037:antagonists & inhibitors; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008555:Melitten / Q000494:pharmacology; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology
chemical_list: D001514:Bee Venoms; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D008555:Melitten; D009569:Nitric Oxide; D002351:Carrageenan; D008156:Luciferases; D015232:Dinoprostone; D002118:Calcium