Determinación de la inhibición de la actividad enzimática del veneno de dos especies de serpientes (Bothrops spp.) por Cordyline fruticosa.

Abstract

This research is aimed at determining an inhibitory activity originated from Cordyline fruticosa plant extract based on Bothrops atrox and Bothrops asper poisonous snake species in the enzyme hyaluronidase, phospholipase, toxin activities on casein, protein, and coagulation. For assessing the enzyme inhibition, a toxin was used in an individual concentration of approximately 20 mg dissolved into a potassium phosphate monobasic buffer and potassium phosphate dibasic, so this treatment is similar to the human organism real conditions. The substrates used to perform the different tests were: hyaluronic acid, 45% egg yolk lipoprotein emulsion, casein, and fibrinogen. To determine the activity of hyaluronidase from toxin, hyaluronic acid was used as a substrate determining the turbidity reduction with cetyl trimethyl ammonium bromide. In the case of phospholipase, a 45% egg yolk lipoprotein emulsion with Tris-HCl buffer and calcium chloride at a pH of 7.4 was used, measuring the coagulation time. In order to determine the casenoylitic activity, in a Tris-HCl buffer was used at a pH of 8.5, precipitating it with trichloroacetic acid before reading its absorbance. Whereas, to establish the coagulation activity, fibrinogen in a Tris-HCl buffer was used at a pH of 7.4, incubating it and then measuring the coagulation time. Once the results of each test were obtained, the samples obtained were compared with polyvalent antiofidic serum and it was obtained that, hyaluronidase is mostly inhibited with the extract of the dried leaves of Cordyline fruticosa in a 4.46% more with respect to the anti-phytic serum It should be noted that the caseinolytic activity is inhibited at very low concentrations, while the activity of phospholipase A2 and the coagulant activity is inhibited at very low concentrations and there is a difference in slowdown in coagulation times. Statistical analyses showed that, in the inhibition of phospholipase, so that caseinolytic and coagulation activity have the same effect as the anti-phytic serum, whereas, the inhibition of hyaluronidase by Cordyline fruticosa has a significant difference with respect to the antophytic serum This research generates expectations for future investigations on inhibitory effects of the enzymes present in the poison using extract of this studied plant since it has inhibitory potential.