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Primary targets of the sesquiterpene lactone deoxymikanolide on Trypanosoma cruzi.
Phytomedicine. 2019 Mar 15; 56:27-34.P

Abstract

BACKGROUND

Deoxymikanolide is a sesquiterpene lactone isolated from Mikania micrantha and M. variifolia which, has previously demonstrated in vitro activity on Trypanosoma cruzi and in vivo activity on an infected mouse model.

PURPOSE

Based on these promising findings, the aim of this study was to investigate the mechanism of action of this compound on different parasite targets.

METHODS

The interaction of deoxymikanolide with hemin was examined under reducing and non- reducing conditions by measuring modifications in the Soret absorption band of hemin; the thiol interaction was determined spectrophotometrically through its reaction with 5,5'-dithiobis-2-nitrobenzoate in the presence of glutathione; activity on the parasite antioxidant system was evaluated by measuring the activity of the superoxide dismutase and trypanothione reductase enzymes, together with the intracellular oxidative state by flow cytometry. Superoxide dismutase and trypanothione reductase activities were spectrophotometrically tested. Cell viability, phosphatidylserine exposure and mitochondrial membrane potential were assessed by means of propidium iodide, annexin-V and rhodamine 123 staining, respectively; sterols were qualitatively and quantitatively tested by TLC; ultrastructural changes were analyzed by transmission electron microscopy. Autophagic cells were detected by staining with monodansylcadaverine.

RESULTS

Deoxymikanolide decreased the number of reduced thiol groups within the parasites, which led to their subsequent vulnerability to oxidative stress. Treatment of the parasites with the compound produced a depolarization of the mitochondrial membrane even though the plasma membrane permeabilization was not affected. Deoxymikanolide did not affect the intracellular redox state and so the mitochondrial dysfunction produced by this compound could not be attributed to ROS generation. The antioxidant defense system was affected by deoxymikanolide at twenty four hours of treatment, when both an increased oxidative stress and decreased activity of superoxide dismutase and trypanothione reductase (40 and 60% respectively) were observed. Both the oxidative stress and mitochondrial dysfunction induce parasite death by apoptosis and autophagy.

CONCLUSION

Based on our results, deoxymikanolide would exert its anti-T cruzi activity as a strong thiol blocking agent and by producing mitochondrial dysfunction.

Authors+Show Affiliations

CONICET - Universidad de Buenos Aires. Centro de Investigaciones sobre Porfirinas y Porfirias, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina.Universidad de Buenos Aires, Cátedra de Farmacognosia, Facultad de Farmacia y Bioquímica, Junín 956 2 P (1113). Buenos Aires, Argentina.Facultad de Ciencias Médicas, Instituto de Histología y Embriología "Dr. Mario H. Burgos", Universidad Nacional de Cuyo-CONICET, Mendoza CC 56 (5500), Argentina.Laboratorio de Inmunología y Desarrollo de Vacunas, Instituto de Medicina y Biología Experimental de Cuyo, Av. Ruiz Leal s/n Parque General San Martín, Mendoza CP 5500, Argentina.CONICET - Universidad de Buenos Aires. Instituto de Química y Metabolismo del Fármaco - CONICET (IQUIMEFA), Junín 956 2 P (1113), Buenos Aires, Argentina.Facultad de Ciencias Médicas, Instituto de Histología y Embriología "Dr. Mario H. Burgos", Universidad Nacional de Cuyo-CONICET, Mendoza CC 56 (5500), Argentina.Universidad de Buenos Aires, Cátedra de Farmacognosia, Facultad de Farmacia y Bioquímica, Junín 956 2 P (1113). Buenos Aires, Argentina; CONICET - Universidad de Buenos Aires. Instituto de Química y Metabolismo del Fármaco - CONICET (IQUIMEFA), Junín 956 2 P (1113), Buenos Aires, Argentina. Electronic address: vsulsen@ffyb.uba.ar.CONICET - Universidad de Buenos Aires. Centro de Investigaciones sobre Porfirinas y Porfirias, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina. Electronic address: elombardo@qb.fcen.uba.ar.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30668348

Citation

Puente, Vanesa, et al. "Primary Targets of the Sesquiterpene Lactone Deoxymikanolide On Trypanosoma Cruzi." Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, vol. 56, 2019, pp. 27-34.
Puente V, Laurella LC, Spina RM, et al. Primary targets of the sesquiterpene lactone deoxymikanolide on Trypanosoma cruzi. Phytomedicine. 2019;56:27-34.
Puente, V., Laurella, L. C., Spina, R. M., Lozano, E., Martino, V. S., Sosa, M. A., Sülsen, V. P., & Lombardo, E. (2019). Primary targets of the sesquiterpene lactone deoxymikanolide on Trypanosoma cruzi. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, 56, 27-34. https://doi.org/10.1016/j.phymed.2018.10.015
Puente V, et al. Primary Targets of the Sesquiterpene Lactone Deoxymikanolide On Trypanosoma Cruzi. Phytomedicine. 2019 Mar 15;56:27-34. PubMed PMID: 30668348.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Primary targets of the sesquiterpene lactone deoxymikanolide on Trypanosoma cruzi. AU - Puente,Vanesa, AU - Laurella,Laura C, AU - Spina,Renata M, AU - Lozano,Esteban, AU - Martino,Virginia S, AU - Sosa,Miguel A, AU - Sülsen,Valeria P, AU - Lombardo,Elisa, Y1 - 2018/10/11/ PY - 2018/08/14/received PY - 2018/10/08/revised PY - 2018/10/10/accepted PY - 2019/1/23/pubmed PY - 2019/6/18/medline PY - 2019/1/23/entrez KW - DCF: Dichlorofluorescein KW - DCIP: 2,6-dichlorophenolindophenol, DMSO: dimethyl sulfoxide, DTNB: 5,5′-dithiobis-2-nitrobenzoate, GSH: glutathione, PBS: phosphate buffered saline, PI: propidium iodide, Rh123: rhodamine 123, ROS: reactive oxygen species, SCR: succinate cytochrome c reductase, SOD: superoxide dismutase, TryR: trypanothione reductase KW - Deoxymikanolide KW - H2DCFDA: 2′,7′-dichlorodihydrofluorescein diacetate KW - Mitochondrial dysfunction KW - Oxidative stress KW - Sesquiterpene lactone KW - Trypanosoma cruzi KW - Ultraestructural damage SP - 27 EP - 34 JF - Phytomedicine : international journal of phytotherapy and phytopharmacology JO - Phytomedicine VL - 56 N2 - BACKGROUND: Deoxymikanolide is a sesquiterpene lactone isolated from Mikania micrantha and M. variifolia which, has previously demonstrated in vitro activity on Trypanosoma cruzi and in vivo activity on an infected mouse model. PURPOSE: Based on these promising findings, the aim of this study was to investigate the mechanism of action of this compound on different parasite targets. METHODS: The interaction of deoxymikanolide with hemin was examined under reducing and non- reducing conditions by measuring modifications in the Soret absorption band of hemin; the thiol interaction was determined spectrophotometrically through its reaction with 5,5'-dithiobis-2-nitrobenzoate in the presence of glutathione; activity on the parasite antioxidant system was evaluated by measuring the activity of the superoxide dismutase and trypanothione reductase enzymes, together with the intracellular oxidative state by flow cytometry. Superoxide dismutase and trypanothione reductase activities were spectrophotometrically tested. Cell viability, phosphatidylserine exposure and mitochondrial membrane potential were assessed by means of propidium iodide, annexin-V and rhodamine 123 staining, respectively; sterols were qualitatively and quantitatively tested by TLC; ultrastructural changes were analyzed by transmission electron microscopy. Autophagic cells were detected by staining with monodansylcadaverine. RESULTS: Deoxymikanolide decreased the number of reduced thiol groups within the parasites, which led to their subsequent vulnerability to oxidative stress. Treatment of the parasites with the compound produced a depolarization of the mitochondrial membrane even though the plasma membrane permeabilization was not affected. Deoxymikanolide did not affect the intracellular redox state and so the mitochondrial dysfunction produced by this compound could not be attributed to ROS generation. The antioxidant defense system was affected by deoxymikanolide at twenty four hours of treatment, when both an increased oxidative stress and decreased activity of superoxide dismutase and trypanothione reductase (40 and 60% respectively) were observed. Both the oxidative stress and mitochondrial dysfunction induce parasite death by apoptosis and autophagy. CONCLUSION: Based on our results, deoxymikanolide would exert its anti-T cruzi activity as a strong thiol blocking agent and by producing mitochondrial dysfunction. SN - 1618-095X UR - https://www.unboundmedicine.com/medline/citation/30668348/Primary_targets_of_the_sesquiterpene_lactone_deoxymikanolide_on_Trypanosoma_cruzi_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0944-7113(18)30536-1 DB - PRIME DP - Unbound Medicine ER -