Tags

Type your tag names separated by a space and hit enter

The antifungal activity and mechanisms of action of quantified extracts from berries, leaves and roots of Phytolacca tetramera.
Phytomedicine. 2019 Jul; 60:152884.P

Abstract

BACKGROUND

Phytolacca tetramera is an endemic plant from Argentina that is currently at serious risk because its environment is subjected to a high anthropic impact. A previous study has shown that berry extracts obtained from this plant display antifungal activity against multiple human-pathogenic fungi when tested with a non-standardized method. Further evidences of the antifungal properties of other parts of the plant and studies of mechanism of antifungal action of the antifungal chemically characterized extracts are required.

PURPOSE

This study aimed to gain further evidence of the antifungal activity of P. tetramera berry, leaf and root extracts in order to find the most active extract to be developed as an Herbal Medicinal Antifungal Product. The medicinal usefulness of P. tetramera extracts as antifungal agents will serve as an important support to create concience and carry out actions tending to the preservation of this threatened species and its environment.

MATERIALS AND METHODS

Chemical analysis of all P. tetramera extracts, including quantitation of selected markers, was performed through UHPLC-ESI-MS/MS and UPLC-ESI-MS techniques according to the European Medicines Agency (EMA). The antifungal activity of the quantified extracts was tested with the standardized CLSI microbroth dilution method against Candida spp. Antifungal mechanisms of the most active extract were studied by examination of morphological changes by phase-contrast and fluorescence microscopies and both, cellular and enzymatic assays targeting either the fungal membrane or the cell wall.

RESULTS

The antifungal activity of twelve P. tetramera extracts was tested against Candida albicans and Candida glabrata. The dichloromethane extract from berries (PtDEb) showed the best activity. Phytolaccagenin (PhytG) and phytolaccoside B (PhytB) were selected as the main active markers for the antifungal P. tetramera extracts. The quantitation of these active markers in all extracts showed that PtDEb possessed the highest amount of PhytG and PhytB. Finally, studies on the mechanism of antifungal action showed that the most active PtDEb extract produces morphological changes compatible with a damage of the cell wall and/or the plasma membrane. Cellular and enzymatic assays showed that PtDEb would not damage the fungal cell wall by itself, but would alter the plasma membrane. In agreement, PtDEb was found to bind to ergosterol, the main sterol of the fungal plasma membrane.

CONCLUSION

Studies of the anti-Candida activity of P. tetramera extracts led to the selection of PtDEb as the most suitable extract, confirming the antifungal properties of the threatened species P. tetramera. The new data give a valuable reason for the definitive protection of this sp. and its natural environment thus allowing further studies for the future development of an Herbal Medicinal Antifungal Product.

Authors+Show Affiliations

Área Farmacognosia, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR), Suipacha 531, 2000 Rosario, Argentina.Área Farmacognosia, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR), Suipacha 531, 2000 Rosario, Argentina.Natural Products Chemistry Department, & State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, China; School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, CMU-Rue Michel-Servet 1, CH-1206 Geneva 4, Switzerland.School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, CMU-Rue Michel-Servet 1, CH-1206 Geneva 4, Switzerland.Instituto de Biología Funcional y Genómica y Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Científicas (CSIC)/Universidad de Salamanca, 37007 Salamanca, Spain.Instituto de Biología Funcional y Genómica y Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Científicas (CSIC)/Universidad de Salamanca, 37007 Salamanca, Spain.Fundación MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento 34, E-18016 Granada, Spain.Fundación MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento 34, E-18016 Granada, Spain.Fundación MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento 34, E-18016 Granada, Spain.Área Farmacognosia, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR), Suipacha 531, 2000 Rosario, Argentina. Electronic address: szaabgil@citynet.net.ar.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30922815

Citation

Butassi, Estefanía, et al. "The Antifungal Activity and Mechanisms of Action of Quantified Extracts From Berries, Leaves and Roots of Phytolacca Tetramera." Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, vol. 60, 2019, p. 152884.
Butassi E, Svetaz LA, Zhou S, et al. The antifungal activity and mechanisms of action of quantified extracts from berries, leaves and roots of Phytolacca tetramera. Phytomedicine. 2019;60:152884.
Butassi, E., Svetaz, L. A., Zhou, S., Wolfender, J. L., Cortés, J. C. G., Ribas, J. C., Díaz, C., Palacio, J. P., Vicente, F., & Zacchino, S. A. (2019). The antifungal activity and mechanisms of action of quantified extracts from berries, leaves and roots of Phytolacca tetramera. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, 60, 152884. https://doi.org/10.1016/j.phymed.2019.152884
Butassi E, et al. The Antifungal Activity and Mechanisms of Action of Quantified Extracts From Berries, Leaves and Roots of Phytolacca Tetramera. Phytomedicine. 2019;60:152884. PubMed PMID: 30922815.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The antifungal activity and mechanisms of action of quantified extracts from berries, leaves and roots of Phytolacca tetramera. AU - Butassi,Estefanía, AU - Svetaz,Laura A, AU - Zhou,Shuaizhen, AU - Wolfender,Jean-Luc, AU - Cortés,Juan C G, AU - Ribas,Juan C, AU - Díaz,Caridad, AU - Palacio,José Pérez-Del, AU - Vicente,Francisca, AU - Zacchino,Susana A, Y1 - 2019/03/16/ PY - 2019/01/23/received PY - 2019/02/25/revised PY - 2019/03/09/accepted PY - 2019/3/30/pubmed PY - 2019/12/18/medline PY - 2019/3/30/entrez KW - Candida albicans KW - Candida glabrata KW - Phytolacca tetramera KW - Phytolaccagenin KW - Phytolaccoside B KW - Quantified extracts SP - 152884 EP - 152884 JF - Phytomedicine : international journal of phytotherapy and phytopharmacology JO - Phytomedicine VL - 60 N2 - BACKGROUND: Phytolacca tetramera is an endemic plant from Argentina that is currently at serious risk because its environment is subjected to a high anthropic impact. A previous study has shown that berry extracts obtained from this plant display antifungal activity against multiple human-pathogenic fungi when tested with a non-standardized method. Further evidences of the antifungal properties of other parts of the plant and studies of mechanism of antifungal action of the antifungal chemically characterized extracts are required. PURPOSE: This study aimed to gain further evidence of the antifungal activity of P. tetramera berry, leaf and root extracts in order to find the most active extract to be developed as an Herbal Medicinal Antifungal Product. The medicinal usefulness of P. tetramera extracts as antifungal agents will serve as an important support to create concience and carry out actions tending to the preservation of this threatened species and its environment. MATERIALS AND METHODS: Chemical analysis of all P. tetramera extracts, including quantitation of selected markers, was performed through UHPLC-ESI-MS/MS and UPLC-ESI-MS techniques according to the European Medicines Agency (EMA). The antifungal activity of the quantified extracts was tested with the standardized CLSI microbroth dilution method against Candida spp. Antifungal mechanisms of the most active extract were studied by examination of morphological changes by phase-contrast and fluorescence microscopies and both, cellular and enzymatic assays targeting either the fungal membrane or the cell wall. RESULTS: The antifungal activity of twelve P. tetramera extracts was tested against Candida albicans and Candida glabrata. The dichloromethane extract from berries (PtDEb) showed the best activity. Phytolaccagenin (PhytG) and phytolaccoside B (PhytB) were selected as the main active markers for the antifungal P. tetramera extracts. The quantitation of these active markers in all extracts showed that PtDEb possessed the highest amount of PhytG and PhytB. Finally, studies on the mechanism of antifungal action showed that the most active PtDEb extract produces morphological changes compatible with a damage of the cell wall and/or the plasma membrane. Cellular and enzymatic assays showed that PtDEb would not damage the fungal cell wall by itself, but would alter the plasma membrane. In agreement, PtDEb was found to bind to ergosterol, the main sterol of the fungal plasma membrane. CONCLUSION: Studies of the anti-Candida activity of P. tetramera extracts led to the selection of PtDEb as the most suitable extract, confirming the antifungal properties of the threatened species P. tetramera. The new data give a valuable reason for the definitive protection of this sp. and its natural environment thus allowing further studies for the future development of an Herbal Medicinal Antifungal Product. SN - 1618-095X UR - https://www.unboundmedicine.com/medline/citation/30922815/The_antifungal_activity_and_mechanisms_of_action_of_quantified_extracts_from_berries_leaves_and_roots_of_Phytolacca_tetramera_ DB - PRIME DP - Unbound Medicine ER -