title: Morphological and biochemical evidence of blood vessel damage and fibrinogenolysis triggered by brown spider venom.
abstract: The venom of the brown spider is remarkable because it causes dermonecrotic injury, hemorrhagic problems, hemolysis, platelet aggregation and renal failure. The mechanism by which the venom causes hemorrhagic disorders is poorly understood. Rabbits intradermally exposed to the venom showed a local hemorrhage starting 1 h after inoculation and reaching maximum activity between 2 and 3 days. Biopsies examined by light and transmission electron microscopy showed subendothelial blebs, vacuoles and endothelial cell membrane degeneration in blood vessels, plasma exudation into connective tissue, and fibrin and thrombus formation within blood vessels. Loxosceles intermedia venom incubated with fibrinogen partially degrades Aalpha and Bbeta chains of intact fibrinogen, and significantly cleaves all Aalpha, Bbeta and gamma chains when they were separated or when fibrinogen is denatured by boiling. Proteolytic kinetic studies showed that the Aalpha chain is more susceptible to venom hydrolysis than the Bbeta chain. The fibrinogenolysis is blocked by ethylenediamine tetraacetic acid and 1,10-phenanthroline, but not by other protease inhibitors. Human plasma incubated with the venom had coagulation parameters such as prothrombin time, activated partial thromboplastin time and thrombin time increased. Through molecular sieve chromatography, we isolated a venom toxin of 30 kDa with fibrinogenolytic activity. We propose that the local and systemic hemorrhagic disorders evoked in loxoscelism are consequences of direct venom fibrinogenolysis together with cytotoxicity to subendothelial structures and endothelial cells in blood vessels.
authors.affiliation: Department of Cell Biology, Federal University of Parana, Jardim das Americas, 81531-990 Curitiba, Parana, Brazil.
authors.forename: V C
authors.initials: VC
authors.lastname: Zanetti
authors.forename: R B
authors.initials: RB
authors.lastname: da Silveira
authors.forename: J L
authors.initials: JL
authors.lastname: Dreyfuss
authors.forename: J
authors.initials: J
authors.lastname: Haoach
authors.forename: O C
authors.initials: OC
authors.lastname: Mangili
authors.forename: S S
authors.initials: SS
authors.lastname: Veiga
authors.forename: W
authors.initials: W
authors.lastname: Gremski
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D001808:Blood Vessels / Q000187:drug effects* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D005340:Fibrinogen / Q000187:drug effects* / Q000378:metabolism; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D013111:Spider Venoms / Q000032:analysis / Q000494:pharmacology / Q000633:toxicity*; D014118:Toxins, Biological / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011480:Protease Inhibitors; D013111:Spider Venoms; D014118:Toxins, Biological; D005340:Fibrinogen