title: Inhibition of venom serine proteinase and metalloproteinase activities by Renealmia alpinia (Zingiberaceae) extracts: comparison of wild and in vitro propagated plants.
abstract: BACKGROUND
  The plant Renealmia alpinia has been used in folk medicine to treat snakebites in the northwest region of Colombia. In addition, it has been shown to neutralize edema-forming, hemorrhagic, lethal, and defibrin(ogen)ating activities of Bothrops asper venom. In this work, extracts of Renealmia alpinia obtained by micropropagation (in vitro) and from specimens collected in the wild were tested and compared in their capacity to inhibit enzymatic and toxic activities of a snake venom metalloproteinase isolated from Bothrops atrox (Batx-I) venom and a serine proteinase (Cdc SII) from Crotalus durissus cumanensis venom.
  
  
  METHODS
  We have investigated the inhibition capacity of Renealmia alpinia extracts on enzymatic and toxic actions of isolated toxins, a metalloproteinase and a serine proteinase. The protocols investigated included inhibition of proteolytic activity on azocasein, inhibition of proteolytic activity on fibrinogen, inhibition of pro-coagulant activity, inhibition of hemorrhagic activity and inhibition of edema-forming activity.
  
  
  RESULTS
  Colorimetric assays detected the presence of terpenoids, flavonoids, tannins and coumarins in Renealmia alpinia extracts. Renealmia alpinia extracts inhibited the enzymatic, hemorrhagic and fibrinogenolytic activities of Batx-I. Extracts also inhibited coagulant, defibrin(ogen)ating and edema-forming activities of Cdc SII. Results highlight that Renealmia alpinia in vitro extract displayed comparable inhibitory capacity on venom proteinases that Renealmia alpinia wild extract. No alteration was observed in the electrophoretic pattern of venom proteinases after incubation with Renealmia alpinia extracts, thus excluding proteolytic degradation or protein denaturation/precipitation as a mechanism of inhibition.
  
  
  CONCLUSIONS
  Our results showed that Renealmia alpinia wild and in vitro extracts contain compounds that neutralize metallo- and serine proteinases present in snake venoms. The mechanism of inhibition is not related to proteolytic degradation of the enzymes nor protein aggregation, but is likely to depend on molecular interactions of secondary metabolites in the plant with these venom proteinases.
authors.affiliation: Departamento de Farmacia, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia. arleycamilo@gmail.com
authors.forename: Arley Camilo
authors.initials: AC
authors.lastname: Patiño
authors.forename: Dora María
authors.initials: DM
authors.lastname: Benjumea
authors.forename: Jaime Andrés
authors.initials: JA
authors.lastname: Pereañez
mesh_terms: D000818:Animals; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D004487:Edema / Q000517:prevention & control; D005340:Fibrinogen / Q000037:antagonists & inhibitors; D006470:Hemorrhage / Q000517:prevention & control; D045726:Metalloproteases / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D051379:Mice; D010936:Plant Extracts / Q000494:pharmacology*; D057057:Serine Proteases / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D030521:Zingiberaceae
chemical_list: D003435:Crotalid Venoms; D010936:Plant Extracts; D015842:Serine Proteinase Inhibitors; D005340:Fibrinogen; C554059:Batx-I, Bothrops atrox; D045726:Metalloproteases; D057057:Serine Proteases
keywords: Anti-venom activity; Metalloproteinase; Micropropagated plants; Renealmia alpinia; Serine proteinase; Snake venoms