title: The complement system is involved in acute inflammation but not in the hemorrhage produced by a Bothrops atrox snake venom low molecular mass proteinase.
abstract: Low molecular weight hemorrhagins were purified from crude Bothrops atrox snake venom by gel filtration followed by ionic strength chromatography. The protein fractions obtained, designated HI-1 to HI-8, contained proteins with molecular masses lower than 30 kDa. HI-5, the most representative among of these fractions, exhibited, in vitro, proteolytic and C inactivating properties, as analyzed by proteolysis of a protein substrate, and C system consumptive activities as assayed by reduction of the hemolytic C activity in normal human serum and by cleavage of partially purified component C3. HI-5 hemorrhagin injected i.m. into C-sufficient BALB/c mice induced a local inflammation characterized by edema, accumulation of polymorphonuclear leucocytes (PMN) and hemorrhage. In contrast, when injected into BALB/c mice previously C-depleted, the number of PMN per tissue section, but not hemorrhage, was significantly reduced (129.668 +/- 31.341 cells per microscopic field) as compared with the control C-sufficient mice (812.168 +/- 111.194 cells per microscopic field). The observations were confirmed by using C5-deficient mice instead of C-depleted mice. The average number of PMN per tissue section in C5-defficient A/J mice was 72.666 +/- 19.416 cells per microscopic field. These data indicate that the C system is involved in PMN accumulation, but not in the hemorrhage, at the local induced lesions by low molecular mass B. atrox hemorrhagins. HI-5 apparently is not contaminated with other direct or indirect inflammation mediators, PMN accumulation and hemorrhage, however, an independent phenomenon, could be mediated by the same hemorrhagin proteinase domain.
authors.affiliation: Laboratório de Biologia do Reconhecer, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego, 2000, Campos dos Goytacazes, Rio de Janeiro 28015-620, Brazil.
authors.forename: Flávia G
authors.initials: FG
authors.lastname: Rodrigues
authors.forename: Jorge H
authors.initials: JH
authors.lastname: Petretski
authors.forename: Milton M
authors.initials: MM
authors.lastname: Kanashiro
authors.forename: Luciana
authors.initials: L
authors.lastname: Lemos
authors.forename: Wilmar Dias
authors.initials: WD
authors.lastname: da Silva
authors.forename: Thereza L
authors.initials: TL
authors.lastname: Kipnis
mesh_terms: D000818:Animals; D017837:Bothrops; D003182:Complement C5 / Q000276:immunology; D015941:Complement Hemolytic Activity Assay; D003165:Complement System Proteins / Q000172:deficiency / Q000502:physiology*; D003435:Crotalid Venoms / Q000201:enzymology* / Q000633:toxicity; D004487:Edema / Q000139:chemically induced; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D007249:Inflammation / Q000139:chemically induced* / Q000473:pathology; D007273:Injections, Intramuscular; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000633:toxicity*; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000473:pathology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D044967:Serum / Q000378:metabolism; D013045:Species Specificity
chemical_list: D003182:Complement C5; D003435:Crotalid Venoms; D003165:Complement System Proteins; D008666:Metalloendopeptidases