title: The potential of aqueous extracts of Bellucia dichotoma Cogn. (Melastomataceae) to inhibit the biological activities of Bothrops atrox venom: A comparison of specimens collected in the states of Pará and Amazonas, Brazil.
abstract: BACKGROUND
  The effectiveness of aqueous extract of Bellucia dichotoma Cogn. (Melastomataceae) specimems collected in Santarém, PA, against some biological activities of Bothrops atrox venom (BaV) has been scientifically proven. Here, we analyzed the components and assessed the anti-snakebite potential of aqueous extracts of bark of B. dichotoma collected in Manaus, AM, (AEBd-MAO) and Santarém, PA, (AEBd-STM), both in Brazil.
  
  
  METHODS
  The phytochemical profiles of the aqueous extracts were identified using thin layer chromatography (TLC), and the concentrations of phenolics were determined by colorimetric assay. The inhibitory potential of the extracts was tested against the phospholipase A2, coagulant and gelatinolytic activities of BaV in vitro and its defibrinating and edema-inducing activities in vivo. Interaction between BaV and the extracts was investigated using SDS-Page electrophoresis and Western blotting. Extract cytotoxicity and antioxidant potential were assessed using the human fibroblast cell line MRC-5 and the DPPH assay in cell culture, respectively.
  
  
  RESULTS
  While there was no difference between the phytochemical profiles of the extracts, AEBd-MAO had higher concentrations of total phenolics, total tannins and hydrolysable tannins. The extracts inhibited 100% of the phospholipase and coagulant activity of BaV when pre-incubated. Without pre-incubation, however, there was no reduction in phospholipase activity, although significant inhibition of coagulant activity was observed. In the doses used in folk medicine, without pre-incubation, both extracts inhibited 100% of the coagulant activity of BaV. In vivo, the extracts were unable to inhibit the defibrinating activity of the venom but were effective in inhibiting its edema-inducing activity. In the profiles of the extracts pre-incubated with BaV, not all the protein bands revealed by SDS-PAGE and Western blot were observed. Both extracts had a high antioxidant potential and neither had a cytotoxic effect.
  
  
  CONCLUSIONS
  Although the concentrations of phenolics in each extract were different, the anti-snakebite potential was similar for the concentrations of extract tested. Our findings are of importance for the quality control of this raw material, which, once tested in accordance with Brazilian National Health Surveillance Agency recommendations, may be suitable for use as a phytomedicine to complement treatment of the local effects induced by Bothrops venoms.
authors.affiliation: Programa Multi-institucional de Pós-graduação em Biotecnologia, 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, Manaus, AM, Brazil. Electronic address: mouraovm@yahoo.com.br.
authors.forename: Valéria Mourão
authors.initials: VM
authors.lastname: de Moura
authors.affiliation: Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil.
authors.forename: Luana Yamille Andrade
authors.initials: LY
authors.lastname: de Souza
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: Noranathan
authors.initials: N
authors.lastname: da Costa Guimarães
authors.affiliation: Programa Multi-institucional de Pós-graduação em Biotecnologia, 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, Manaus, AM, Brazil.
authors.forename: Ilia Gilmara Carvalho
authors.initials: IG
authors.lastname: Dos Santos
authors.affiliation: Laboratório de Atividade Biológica - Biophar,Biophar, Faculdade de Ciências Farmacêuticas, Universidade Federal do Amazonas, AM, Brazil.
authors.forename: Patrícia Danielle Oliveira
authors.initials: PD
authors.lastname: de Almeida
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: Ricardo Bezerra
authors.initials: RB
authors.lastname: de Oliveira
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: RH
authors.lastname: Mourão
authors.affiliation: Programa Multi-institucional de Pós-graduação em Biotecnologia, 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, Manaus, AM, Brazil.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Dos-Santos
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology / Q000627:therapeutic use; D000925:Anticoagulants / Q000494:pharmacology / Q000627:therapeutic use; D000975:Antioxidants / Q000494:pharmacology / Q000627:therapeutic use; D000997:Antivenins / Q000494:pharmacology / Q000627:therapeutic use; D001777:Blood Coagulation / Q000187:drug effects; D017837:Bothrops; D001938:Brazil; D002460:Cell Line; D003435:Crotalid Venoms / Q000633:toxicity*; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy; D006801:Humans; D008297:Male; D035322:Melastomataceae; D051379:Mice; D054467:Phospholipases A2 / Q000378:metabolism; D064209:Phytochemicals / Q000032:analysis; D008517:Phytotherapy; D024301:Plant Bark; D010936:Plant Extracts / Q000032:analysis / Q000494:pharmacology / Q000627:therapeutic use; D012909:Snake Bites / Q000188:drug therapy
chemical_list: D000893:Anti-Inflammatory Agents; D000925:Anticoagulants; D000975:Antioxidants; D000997:Antivenins; D003435:Crotalid Venoms; D064209:Phytochemicals; D010936:Plant Extracts; D054467:Phospholipases A2
keywords: Anti-snakebite plants; Bellucia dichotoma; Bothrops; Folk medicine; Snakebite