title: Occurrence of sulfated fucose branches in fucosylated chondroitin sulfate are essential for the polysaccharide effect preventing muscle damage induced by toxins and crude venom from Bothrops jararacussu snake.
abstract: Snake envenoming is an important public health problem around the world, particularly in tropics. Beyond deaths, morbidity induced by snake venoms, such as myotoxicity, is of pivotal consequence to population. Bothrops jararacussu is the main venomous snake in southeast region of Brazil, and particularly presents strong myotoxic effect. The only available therapy, antibothropic antivenom, poorly affects venom-induced myotoxicity. The aim of this study is to assess the ability of fucosylated chondroitin sulfate (fucCS), a glycosaminoglycan with anticoagulant and antithrombotic properties, and its derivatives to inhibit toxic activities of B. jararacussu crude venom and its isolated toxins, named bothropstoxins (BthTX-I and BthTX-II). The in vitro myotoxic activities induced by crude venom, by BthTX-I alone and by toxins together were abolished by fucCS. Carboxyl reduction (fucCS-CR) kept this ability whereas defucosilation (defucCS) abrogates myoprotection. We observed the same pattern in the response of these polysaccharides in antagonizing the increase in plasma creatine kinase (CK) levels, the reduction of skeletal muscle CK content and the rise of myeloperoxidase (MPO) activity induced by crude venom and isolated toxins. FucCS inhibited edematogenic activity and partially prevented the reduction of total leukocytes in blood when pre-incubated with crude venom. Furthermore, the venom procoagulant effect was completely antagonized by increasing concentrations of fucCS, although this polyanion could stop neither the tail bleeding nor the skin hemorrhage induced by Bothrops jararaca venom. The B. jararacussu phospholipase, hyaluronidase, proteolytic and collagenase activities were inhibited in vitro. The results suggest that fucCS could be able to interact with both toxins, and it is able to inhibit BthTX-II phospholipase activity. Light microscopy of extensor digitorum longus muscle (EDL) muscle showed myoprotection by fucCS, once necrotic areas, edema and inflammatory cells were all decreased as compared to venom injection alone. Altogether, data show that fucCS was able to inhibit myotoxicity and inflammation induced by B. jararacussu venom and its phospholipase toxins, BthTX-I and BthTX-II. Thus, fucosylated chondroitin sulfate is a new polyanion with potential to be used as an adjuvant in the treatment of snakebites in the future.
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Marcos
authors.initials: M
authors.lastname: Monteiro-Machado
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Marcelo A
authors.initials: MA
authors.lastname: Tomaz
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Roberto J C
authors.initials: RJ
authors.lastname: Fonseca
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Marcelo A
authors.initials: MA
authors.lastname: Strauch
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Bruno L
authors.initials: BL
authors.lastname: Cons
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Paula A
authors.initials: PA
authors.lastname: Borges
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Fernando C
authors.initials: FC
authors.lastname: Patrão-Neto
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Matheus S
authors.initials: MS
authors.lastname: Tavares-Henriques
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Jhonatha M
authors.initials: JM
authors.lastname: Teixeira-Cruz
authors.affiliation: Departamento de Farmácia e Administração Farmacêutica, Faculdade de Farmácia, UFF, Niterói, RJ, Brazil.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Calil-Elias
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Adélia C O
authors.initials: AC
authors.lastname: Cintra
authors.affiliation: Programa de Pesquisa em Neurociência Básica e Clínica, Instituto de Ciências Biomédicas, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Ana Maria B
authors.initials: AM
authors.lastname: Martinez
authors.affiliation: Laboratório de Tecido Conjuntivo, Hospital Universitário Clementino Fraga Filho and Programa de Glicobiologia, Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Paulo A S
authors.initials: PA
authors.lastname: Mourão
authors.affiliation: Programa de Pós-Graduação em Farmacologia e Química Medicinal, Instituto de Ciências Biomédicas - Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. Electronic address: melo.pa@gmail.com.
authors.forename: Paulo A
authors.initials: PA
authors.lastname: Melo
mesh_terms: D000818:Animals; D017837:Bothrops; D001938:Brazil; D002809:Chondroitin Sulfates / Q000494:pharmacology*; D017364:Collagenases / Q000378:metabolism; D003402:Creatine Kinase / Q000037:antagonists & inhibitors / Q000097:blood; D003435:Crotalid Venoms / Q000633:toxicity*; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy; D005643:Fucose / Q000494:pharmacology*; D054501:Group II Phospholipases A2 / Q000633:toxicity; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D010740:Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism; D012909:Snake Bites / Q000188:drug therapy
chemical_list: D003435:Crotalid Venoms; C517150:fucosylated chondroitin sulfate; C078097:bothropstoxin; D005643:Fucose; D002809:Chondroitin Sulfates; D009195:Peroxidase; D003402:Creatine Kinase; D010740:Phospholipases; D054501:Group II Phospholipases A2; C072031:bothropstoxin II; D006821:Hyaluronoglucosaminidase; D017364:Collagenases
keywords: Bothrops; Bothropstoxin; Fucosylated chondroitin sulfate; Myotoxicity; Snake venom