title: Connarus favosus Planch.: An inhibitor of the hemorrhagic activity of Bothrops atrox venom and a potential antioxidant and antibacterial agent.
abstract: BACKGROUND
  The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites.
  
  
  OBJECTIVE
  To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract.
  
  
  METHODS
  AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography.
  
  
  RESULTS
  The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50)µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked.
  
  
  CONCLUSIONS
  AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.
authors.affiliation: Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil. Electronic address: thaiscwb@yahoo.com.br.
authors.forename: Thaís Pereira da
authors.initials: TPD
authors.lastname: Silva
authors.affiliation: Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Valéria Mourão de
authors.initials: VM
authors.lastname: Moura
authors.affiliation: Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Bioprospecção e Biotecnologia - LABB, Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Maria Carolina Scheffer de
authors.initials: MCS
authors.lastname: Souza
authors.affiliation: Laboratório de Bioprospecção e Biotecnologia - LABB, Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, AM, Brazil.
authors.forename: Vanessa Neves Carvalho
authors.initials: VNC
authors.lastname: Santos
authors.affiliation: Laboratório de Bioprospecção e Biotecnologia - LABB, Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, AM, Brazil.
authors.forename: Kissinara A M Moreira da
authors.initials: KAMMD
authors.lastname: Silva
authors.affiliation: Laboratório de Bioprospecção e Biotecnologia - LABB, Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, AM, Brazil.
authors.forename: Mara Giselle Guimarães
authors.initials: MGG
authors.lastname: Mendes
authors.affiliation: Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Bioprospecção e Biotecnologia - LABB, Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, AM, Brazil.
authors.forename: Cecília Verônica
authors.initials: CV
authors.lastname: Nunez
authors.affiliation: Laboratório de Atividade Biológica - Biophar, Faculdade de Ciências Farmacêuticas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Patrícia Danielle Oliveira de
authors.initials: PDO
authors.lastname: Almeida
authors.affiliation: Laboratório de Atividade Biológica - Biophar, Faculdade de Ciências Farmacêuticas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Emerson Silva
authors.initials: ES
authors.lastname: Lima
authors.affiliation: Programa de Pós-graduação em Recursos Naturais da Amazônia, Laboratório de Bioprospecção e Biologia Experimental, Universidade Federal do Oeste do Pará, Santarém, PA, Brazil.
authors.forename: Rosa Helena Veras
authors.initials: RHV
authors.lastname: Mourão
authors.affiliation: Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Dos-Santos
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D000997:Antivenins / Q000737:chemistry / Q000494:pharmacology*; D017837:Bothrops; D001938:Brazil; D031961:Connaraceae / Q000737:chemistry*; D003435:Crotalid Venoms / Q000008:administration & dosage*; D005260:Female; D006470:Hemorrhage / Q000188:drug therapy* / Q000378:metabolism; D008297:Male; D008519:Medicine, Traditional / Q000379:methods; D051379:Mice; D054467:Phospholipases A2 / Q000378:metabolism; D064209:Phytochemicals / Q000737:chemistry / Q000494:pharmacology; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D012909:Snake Bites / Q000188:drug therapy; D013634:Tannins / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D000975:Antioxidants; D000997:Antivenins; D003435:Crotalid Venoms; D064209:Phytochemicals; D010936:Plant Extracts; D013634:Tannins; D054467:Phospholipases A2
keywords: Anti-snakebite plants; Bothrops atrox; Connarus favosus; Folk medicine; Snakebites