A trypsin inhibitor from <i>Moringa oleifera</i> flower extract is cytotoxic to <i>Trypanosoma cruzi</i> with high selectivity over mammalian cells

Pontual, Emmanuel Viana; Pires-Neto, Divar F; Fraige, Karina; Higino, Taciana Mirely Maciel; Carvalho, Belany E.A.; Alves, Neyla Maria Pereira; Lima, Tânia Andrade; Zingali, Russolina B.; Coelho, Luana Cassandra Breitenbach Barroso; Bolzani, Vanderlan S.; Figueiredo, Regina Célia Bressan Queiroz de; Napoleão, Thiago Henrique

doi:10.1080/14786419.2017.1389932

Cited by 14 publications

(11 citation statements)

References 15 publications

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“…After 24 h, the LC 50 value was 43.5 (26.2-60.9) µg/mL. Similarly, Pontual et al [11] reported an LC 50 value of 41.20 µg/mL for MoFTI on T. cruzi trypomastigotes. In this same work, the authors showed that MoFTI was low in toxicity to murine peritoneal macrophages (50% cytotoxic concentration, CC 50 , of 407.01 µg/mL) and did not interfere with the viability of Vero cells at concentrations up to 500 µg/mL.…”

Section: Resultsmentioning

confidence: 83%

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A Trypsin Inhibitor from Moringa oleifera Flowers Modulates the Immune Response In Vitro of Trypanosoma cruzi-Infected Human Cells

et al. 2020

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Trypanosoma cruzi causes the lethal Chagas disease, which is endemic in Latin America. Flowers of Moringa oleifera (Moringaceae) express a trypsin inhibitor (MoFTI) whose toxicity to T. cruzi trypomastigotes was previously reported. Here, we studied the effects of MoFTI on the viability of human peripheral blood mononuclear cells (PBMCs) as well as on the production of cytokines and nitric oxide (NO) by T. cruzi-infected PBMCs. Incubation with MoFTI (trypsin inhibitory activity: 62 U/mg) led to lysis of trypomastigotes (LC50 of 43.5 µg/mL) but did not affect the viability of PBMCs when tested at concentrations up to 500 µg/mL. A selectivity index > 11.48 was determined. When T. cruzi-infected PBMCs were treated with MoFTI (43.5 or 87.0 µg/mL), the release of the pro-inflammatory cytokine TNF-α and INF-γ, as well as of NO, was stimulated. The release of the anti-inflammatory cytokine IL-10 also increased. In conclusion, the toxicity to T. cruzi and the production of IL-10 by infected PBMCs treated with MoFTI suggest that this molecule may be able to control parasitemia while regulating the inflammation, preventing the progress of Chagas disease. The data reported here stimulate future investigations concerning the in vivo effects of MoFTI on immune response in Chagas disease.

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Section: Resultsmentioning

confidence: 83%

“…The processes of infection of host cells by T. cruzi and the survival of the parasite depend on the activity of important proteases; thus, an imbalance in the activity of these enzymes can cause damages to the parasite [8][9][10]. In this sense, T. cruzi enzymes represent interesting targets for studies of new therapeutic approaches [11].…”

Section: Introductionmentioning

confidence: 99%

A Trypsin Inhibitor from Moringa oleifera Flowers Modulates the Immune Response In Vitro of Trypanosoma cruzi-Infected Human Cells

et al. 2020

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“…In addition, its flowers are consumed and have hypoglycemic, tonic, and diuretic therapeutic actions (Moura et al, 2011;Santos et al, 2012). It also has larvicidal activity against Aedes aegypti and is antiparasitic against Trypanosoma cruzi (Pontual et al, 2012a(Pontual et al, , 2018. Further, it has technological potential in the food industry, due to its caseinolytic activity and milk-coagulating effects (Pontual, et al, 2012b).…”

Section: Discussionmentioning

confidence: 99%

Antimicrobial activity of polypyrrole nanoparticles and aqueous extract ofMoringa oleiferaagainstStaphylococcusspp. carriers of multi-drug efflux system genes isolated from dairy farms

Silva

Barros

Santos

et al. 2020

Journal of Dairy Research

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Our objectives were to identify genes of the multi-drug efflux system and to evaluate the antimicrobial activities of polypyrrole nanoparticles (PPy-NPs) and aqueous extract of Moringa oleifera against Staphylococcus spp. isolated from dairy farms in Northeast Brazil. Initially, 162 Staphylococcus spp. isolates were subjected to in vitro antimicrobial sensitivity tests. Of these, 35 presented antimicrobial multi-drug resistance phenotypes. These 35 isolates were then referred for the detection of norA, norB, norC, msrA, mgrA, tet-38, and lmrS genes, all of which feature in multi-drug efflux systems. In the isolates carrying the genes, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs and Moringa oleifera aqueous extract were determined. In the molecular analysis of the 35 isolates norA, norC, tet-38, and msrA genes were detected and for the other genes norB, lmrS and mgrA there was no amplification. Antimicrobial activity was verified of PPy-NPs and aqueous extract of Moringa oleifera in Staphylococcus spp. carrying multi-drug efflux system genes. We concluded that there are multi-drug efflux system genes present in the Staphylococcus spp. from the agricultural environment in Northeast Brazil, and that aqueous extract of Moringa oleifera and PPy-NPs show bactericidal activity against these isolates.

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“…The PgTI protein spot was excised from the 2dimensional gel and submitted to in-gel tryptic digestion as described by Pontual et al [6]. MS/MS analysis was performed in electrospray ionization quadrupole time-of-flight (ESI-QUAD-TOF) mode and the peptide spectra were compared with sequences of Viridiplantae proteins (NCBInr database) using MASCOT (http://www.matrixscience.com).…”

Section: Mass Spectrometry (Ms) Analysismentioning

confidence: 99%

“…In plants, they can be secondary metabolites, such as flavonoids, or proteins [2]. Proteinaceous PIs isolated from plants have been reported as insecticidal, anti-angiogenic, trypanocidal, and antimicrobial agents [3][4][5][6][7][8]. These inhibitors can affect replication or viability of microbial cells by inhibiting the activity of proteases or damaging the cell wall or plasma membrane leading to alteration of permeability [8].…”

Section: Introductionmentioning

confidence: 99%

PgTI, the First Bioactive Protein Isolated from the Cactus Pilosocereus gounellei, is a Trypsin Inhibitor with Antimicrobial Activity

Filho

Amorim

Lima

et al. 2019

AIR

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Aims: This work aimed to isolate, characterize and evaluate the antimicrobial activity of a trypsin inhibitor (PgTI) from the stem of Pilosocereus gounellei. Place and Duration of Study: Methodology: PgTI was isolated from P. gounellei stem extract by gel filtration and ion exchange chromatographies. The inhibitor was characterized by isoelectric focusing, polyacrylamide gel electrophoresis, tryptic digestion followed by mass spectrometry analysis and for stability towards heating. Antibacterial and antifungal activities were investigated through broth microdilution assays. Viability of the microbial cells was also evaluated by flow cytometry analysis using thiazol orange and propidium iodide. Results: PgTI appeared as a single polypeptide band of 37.1 kDa and isoelectric point (pI) 5.88. The inhibition constant (K i ) for bovine trypsin was 14 nM and mass spectrometry analysis of PgTI did not reveal similarities with other plant proteins. Trypsin inhibitor activity was stable at temperatures up to 50ºC. PgTI inhibited growth of Gram-positive and Gram-negative bacteria (minimal inhibitory concentrations (MIC) from 7.5 to150 µg/mL) with bactericidal activity only against Escherichia coli (minimal bactericidal concentration: 75.0 µg/mL). PgTI also inhibited the growth of Candida krusei (MIC of 60 µg/mL). Flow cytometry confirmed that PgTI did not affect the viability of E. coli and C. krusei cells at the MIC. Conclusion: This is the first report on a bioactive protein purified from P. gounellei, which provides biotechnological value to this cactus.

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A trypsin inhibitor from Moringa oleifera flower extract is cytotoxic to Trypanosoma cruzi with high selectivity over mammalian cells

Cited by 14 publications

References 15 publications

A Trypsin Inhibitor from Moringa oleifera Flowers Modulates the Immune Response In Vitro of Trypanosoma cruzi-Infected Human Cells

A Trypsin Inhibitor from Moringa oleifera Flowers Modulates the Immune Response In Vitro of Trypanosoma cruzi-Infected Human Cells

Antimicrobial activity of polypyrrole nanoparticles and aqueous extract ofMoringa oleiferaagainstStaphylococcusspp. carriers of multi-drug efflux system genes isolated from dairy farms

PgTI, the First Bioactive Protein Isolated from the Cactus Pilosocereus gounellei, is a Trypsin Inhibitor with Antimicrobial Activity

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