title: Mast cell degranulation induced by two phospholipase A2 homologues: dissociation between enzymatic and biological activities.
abstract: Bothropstoxin-I and bothropstoxin-II are phospholipase A2 homologues isolated from Bothrops jararacussu snake venom. The former is devoid of phospholipase A2 activity whereas the latter has very low enzymatic activity. In this study, we have investigated the in vivo (rat paw and skin oedema) and in vitro (mast cell degranulation) inflammatory effects caused by bothropstoxin-I and bothropstoxin-II. Bothropstoxin-I (25-100 microg/paw) and bothropstoxin-II (12.5-50 microg/paw) caused dose-dependent rat paw oedema. The intradermal injection of bothropstoxin-I (0.125-5 microg/site) and bothropstoxin-II (0.125-5 microg/site) into rat skin also resulted in dose-dependent oedema formation. These oedematogenic activities were largely reduced in animals pretreated with the histamine/5-hydroxytryptamine (5-HT) receptor antagonist cyproheptadine (2 mg/kg, i.p. 0.5 h before). Similarly, p-bromophenacyl bromide, a compound known to inhibit phospholipase A2 activity, significantly inhibited rat paw and skin oedema induced by both phospholipase A2 homologues. The polyanion heparin (5 IU/site) significantly reduced the rat skin oedema induced by either bothropstoxin-I or bothropstoxin-II as well as the paw oedema (50 IU/site) induced by the former. When assayed in the rat peritoneal mast cells in vitro, both bothropstoxin-I (10 and 100 microg/ml) and bothropstoxin-II (3 and 10 microg/ml) significantly caused [14C]5-HT release. The [14C]5-HT release caused by these phospholipase A2 homologues were reduced by p-bromophenacyl bromide and heparin (50 IU/ml). Our results indicate that oedema formation induced by bothropstoxin-I and bothropstoxin-II is mostly dependent on in vivo mast cell degranulation. Since heparin greatly reduced the oedematogenic activity of these phospholipase A2 homologues, it is likely that the cationic charge of these substances plays a major role in the mast cell activation. Our results also indicate that p-bromophenacyl bromide may not be a suitable pharmacological tool to investigate the correlation between enzymatic activity and the inflammatory effects of phospholipases A2.
authors.affiliation: Department of Pharmacology, Faculty of Medical Sciences, UNICAMP, Campinas (SP), Brazil. landucci@bestway.com.br
authors.forename: E C
authors.initials: EC
authors.lastname: Landucci
authors.forename: R C
authors.initials: RC
authors.lastname: Castro
authors.forename: M F
authors.initials: MF
authors.lastname: Pereira
authors.forename: A C
authors.initials: AC
authors.lastname: Cintra
authors.forename: J R
authors.initials: JR
authors.lastname: Giglio
authors.forename: S
authors.initials: S
authors.lastname: Marangoni
authors.forename: B
authors.initials: B
authors.lastname: Oliveira
authors.forename: G
authors.initials: G
authors.lastname: Cirino
authors.forename: E
authors.initials: E
authors.lastname: Antunes
authors.forename: G
authors.initials: G
authors.lastname: De Nucci
mesh_terms: D000818:Animals; D015550:Cell Degranulation / Q000187:drug effects*; D003435:Crotalid Venoms / Q000378:metabolism / Q000494:pharmacology*; D004487:Edema / Q000139:chemically induced; D054501:Group II Phospholipases A2; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects*; D010741:Phospholipases A / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D054467:Phospholipases A2; D051381:Rats; D017208:Rats, Wistar; D030162:Reptilian Proteins; D012867:Skin / Q000473:pathology
chemical_list: D003435:Crotalid Venoms; D030162:Reptilian Proteins; C078097:bothropstoxin; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2; C072031:bothropstoxin II