RESUMEN
Enzymatic inhibition by natural compounds may represent a valuable adjuvant in snakebite serum therapy. The objective in this work was to evaluate possible in vitro interactions between vanillic acid and enzymes from Bothrops spp. and Crotalus durissus terrificus venoms, and also suggest a theory as how they interact based on molecular docking. Vanillic acid inhibited the phospholipase activity induced by Bothrops alternatus (â¼25% inhibition); the caseinolytic activity induced by Bothrops atrox (â¼30%), Bothrops jararacussu (â¼44%), and C. d. terrificus (â¼33%); the fibrinogenolysis induced by B. jararacussu, B. atrox, and C. d. terrificus (100%); the serine protease activity induced by Bothrops moojeni (â¼45%) and Bothrops jararaca (â¼66%); the hemolytic activity induced by B. moojeni (â¼26%); the thrombolysis activity induced by B. atrox (â¼30%) and B. jararacussu (â¼20%); and the thrombotic activity induced by C. d. terrificus (â¼8%). The compound was also capable of delaying the coagulation time in citrated plasma by 60, 35, and 75 Sec, when incubated with B. moojeni, B. atrox, and B. jararaca, respectively. The results obtained expand the possibilities for future pharmaceutical use of vanillic acid, considering the high homology degree among human and snake venom phospholipases A2 and proteases (involved in chronic inflammatory diseases). Also, this compound can be used as adjuvant to improve currently available treatments for ophidism victims.
Asunto(s)
Simulación del Acoplamiento Molecular , Péptido Hidrolasas/metabolismo , Inhibidores de Fosfolipasa A2/farmacología , Fosfolipasas A2/metabolismo , Inhibidores de Proteasas/farmacología , Ácido Vanílico/farmacología , Animales , Humanos , Inhibidores de Fosfolipasa A2/química , Inhibidores de Proteasas/química , Serpientes , Ácido Vanílico/químicaRESUMEN
Snake toxins, such as phospholipases A2 and proteases, are used as research tools to evaluate biological activities and to understand physiopathological processes of natural compounds better. In the present study, the phenolic compounds catechin and epicatechin were incubated with snake venoms to evaluate their inhibition against different substrates. Catechin and epicatechin exerted inhibitions between 20% and 95% on the activity of phospholipases A2 present in the venom of Bothrops alternatus. In the hemolytic activity, catechin exerted inhibitions between 20% and 25% in all proportions evaluated on the B. jararacussu venom, whereas epicatechin inhibited 20% of the venom activity. Coagulation induced by B. atrox and B. jararacussu venoms was significantly inhibited by catechin and epicatechin, where the time for coagulation was two to three times higher after previous incubation of the venoms with the compounds. The most significant inhibitions for the proteolytic activity on casein were 17% and 27%, respectively, by both compounds. Catechin inhibited serine protease activity induced by B. atrox venom by 64% and epicatechin by 65%. Regarding B. atrox-induced thrombolysis, catechin exerted 40% inhibition and epicatechin around 30%. The fibrinogen proteolysis was completely inhibited by catechin acting on the B. atrox venom in the proportion of 1:1 and by epicatechin on B. jararacussu venom. Catechin and epicatechin showed promising inhibitory action on proteases and phospholipases A2 . Therefore, these compounds can be explored as an adjuvant for serum therapy or pharmaceutical purposes, once they act on homologous enzymes that are present in humans.
Asunto(s)
Catequina/uso terapéutico , Venenos de Crotálidos/toxicidad , Hemostasis/efectos de los fármacos , Animales , Coagulación Sanguínea/efectos de los fármacos , Bothrops/metabolismo , Catequina/farmacología , Fibrinólisis/efectos de los fármacos , Hemólisis/efectos de los fármacos , Humanos , ProteolisisRESUMEN
The objective of this study was to determine cytotoxic activity, hemolytic activity, and to evaluate the ability of the essential oil from Cinnamodendron dinisii to induce DNA fragmentation of human lymphocytes. The essential oil was obtained by hydrodistillation. Cytotoxic activity was determined by the MTT method. Hemolytic activity was evaluated by spectrophotometric quantification of hemoglobin released by erythrocytes. Damage to lymphocyte DNA molecules was assessed by the Comet assay. The essential oil under study showed high cytotoxic activity on Vero cells (CC50 = 35.72 µg/mL) and induced hemolysis in both hematocrits, besides leading to the oxidation of hemoglobin released. The genotoxic activity of C. dinisii essential oil was also observed, which induced concentration-dependent DNA fragmentation of human lymphocytes and, at 50 µL/mL, it was more active than the positive control. The essential oil from C. dinisii has a toxic action, suggesting a special attention in the application of this oil to health-promoting activities; however, among its components, there are molecules with potential for future application in anticancer therapies.
Asunto(s)
Linfocitos/efectos de los fármacos , Magnoliopsida/química , Aceites Volátiles/farmacología , Animales , Muerte Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Fragmentación del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Hemólisis/efectos de los fármacos , Humanos , Aceites Volátiles/química , Aceites Volátiles/aislamiento & purificación , Relación Estructura-Actividad , Células VeroRESUMEN
The enzyme inhibition by natural and/ or low-cost compounds may represent a valuable adjunct to traditional serotherapy performed in cases of snakebite, mainly with a view to mitigate the local effects of envenoming. The objective of this study was to evaluate possible interactions between vitamins and enzymes that comprise Bothrops atrox and Crotalus durissus terrificus venoms, in vitro. Proteolysis inhibition assays (substrates: azocasein, collagen, gelatin and fibrinogen), hemolysis, coagulation, hemagglutination were carried out using different proportions of vitamins in face of to inhibit minimum effective dose of each venom. The vitamins were responsible for reducing 100% of breaking azocasein by C.d.t. venom, thrombolysis induced by B. atrox and fibrinogenolysis induced by both venoms. It is suggested the presence of interactions between vitamin and the active site of enzymes, for example the interactions between hydrophobic regions present in the enzymes and vitamin E, as well as the inhibitions exercised by antioxidant mechanism.