Abstract
The objective of this study was to investigate the effect on the myeloperoxidase-chlorinating activity in vitro and on neutrophils of Vellozia dasypus Seub., Velloziaceae, and characterize the phenolic active compounds. The methanolic extract, containing the highest phenolic and flavonoid levels, was fractioned on a silica gel column to yield four fractions that inhibited the hypochlorous acid production by myeloperoxidase in a concentration-dependent manner. Chlorogenic acid, caffeic acid, ferulic acid, orientin, and luteolin were identified in the fractions using high-resolution liquid chromatography coupled to diode array detector analysis followed by comparison of their ultraviolet absorption spectra against authentic patterns. Each phenolic compound was assessed against in vitro myeloperoxidase activity and in neutrophil cells. The most active AcOEt-soluble fraction inhibited the enzyme by 95% at the 100 µg/ml concentration, while the methanolic extract inhibited 61% of myeloperoxidase at the same concentration. Caffeic acid, ferulic acid, and luteolin decreased the myeloperoxidase activity with IC50 values around of 4.5 µM. Tyrosine abrogated the inhibitory effect of the compounds suggesting that those compounds blocked myeloperoxidase activity by myeloperoxidase-II accumulation. These compounds reduced the hypochlorous acid formation by neutrophil cells at IC50 10–14 µM. Additional computational studies revealed that these compounds bind at the myeloperoxidase active site by hydrophobic and hydrogen bonding interactions with catalytic residues. The results suggested that caffeic acid, ferulic acid, and luteolin inhibit myeloperoxidase-chlorinating activity through a reversible inhibitory mechanism.
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Aldib I, Soubhye J, Zouaoui Boudjeltia K, Vanhaeverbeek M, Rousseau A, Furtmuller PG, Obinger C, Dufrasne F, Neve J, Van Antwerpen P, Prevost M (2012) Evaluation of new scaffolds of myeloperoxidase inhibitors by rational design combined with high-throughput virtual screening. J Med Chem 55:7208–7218. https://doi.org/10.1021/jm3007245
Almeida VM, Dias ER, Souza BC, Cruz JN, Santos CBR, Leite FHA, Queiroz RF, Branco A (2021) Methoxylated flavonols from Vellozia dasypus Seub ethyl acetate active myeloperoxidase extract: in vitro and in silico assays. J Biomol Struct Dyn 40:7574–7583. https://doi.org/10.1080/07391102.2021.1900916
Alves RJV, Kolbek J (1994) Plant species endemism in savanna vegetation on table mountains (Campo Rupestre) in Brazil. Vegetatio 113:125–139. https://doi.org/10.1007/BF00044230
Ambriz-Pérez DL, Leyva-López N, Gutierrez-Grijalva EP, Heredia JB, Yildiz F (2016) Phenolic compounds: natural alternative in inflammation treatment. A Review Cogent Food Agriculture 2:1. https://doi.org/10.1080/23311932.2015.1131412
Aratani Y (2018) Myeloperoxidase: its role for host defense, inflammation, and neutrophil function. Arch Biochem Biophys 640:47–52. https://doi.org/10.1016/j.abb.2018.01.004
Boufadi Y, Soubhye J, Riazi A, Rousseau A, Vanhaeverbeek M, Nève J, Boudjeltia K, Van Antwerpen P (2014) Characterization and antioxidant properties of six Algerian propolis extracts: ethyl acetate extracts inhibit myeloperoxidase activity. Int J Mol Sci 15:2327–2345. https://doi.org/10.3390/ijms15022327
Branco A, Pereira AS, Cardoso JN, de Aquino Neto FR, Pinto AC, Braz-Filho R (2001) Further lipophilic flavonols in Vellozia graminifolia (Velloziaceae) by high temperature gas chromatography: quick detection of new compounds. Phytochem Anal 12:266–270. https://doi.org/10.1002/pca.590
Branco A, Pinto AC, Ifa DR, Braz Filho R (2002) Two 8C-methylated flavonols from the leaves of Vellozia candida Mikan (Velloziaceae). J Brazil Chem Soc 13:318–323. https://doi.org/10.1590/S0103-50532002000300005
Branco A, Pinto AC, Braz Filho R (2004) Chemical constituents from Vellozia graminifolia (Velloziaceae). An Acad Bras Cien 76:505–518. https://doi.org/10.1590/S0001-37652004000300005
da Silva CG, Carvalho CD, Hamerski L, Castro FA, Alves RJ, Kaiser CR, Eleutherio EC, de Rezende CM (2012) Protective effects of flavonoids and extract from Vellozia kolbekii Alves against oxidative stress induced by hydrogen peroxide in yeast. J Nat Med 66:367–372. https://doi.org/10.1007/s11418-011-0585-z
Davey CA, Fenna RE (1996) 2.3 Å Resolution X-ray crystal structure of the bisubstrate analogue inhibitor salicylhydroxamic acid bound to human myeloperoxidase: a model for a prereaction complex with hydrogen peroxide†,‡. Biochemistry 35:10967–10973. https://doi.org/10.1021/bi960577m
Dias AR, Dias TLMF, Alexandre-Moreira MS (2020) Flavonoid-rich fraction from Pleoma pereirae Melastomataceae effects on calcium oxalate crystallization antioxidant and antinociceptive activies Eur J. Integr Med 35:101095. https://doi.org/10.1016/j.eujim.2020.101095
Figueiredo-Rinhel ASG, de Melo LL, Bortot LO, Santos EOL, Andrade MF, Azzolini A, Kabeya LM, Caliri A, Bastos JK, Lucisano-Valim YM (2017) Baccharis dracunculifolia DC (Asteraceae) selectively modulates the effector functions of human neutrophils. J Pharm Pharmacol 69:1829–1845. https://doi.org/10.1111/jphp.12822
Forbes LV, Sjogren T, Auchere F, Jenkins DW, Thong B, Laughton D, Hemsley P, Pairaudeau G, Turner R, Eriksson H, Unitt JF, Kettle AJ (2013) Potent reversible inhibition of myeloperoxidase by aromatic hydroxamates. J Biol Chem 288:36636–36647. https://doi.org/10.1074/jbc.M113.507756
Franck T, Kohnen S, Grulke S, Neven P, Goutman Y, Peters F, Pirotte B, Deby-Dupont G, Serteyn D (2008) Inhibitory effect of curcuminoids and tetrahydrocurcuminoids on equine activated neutrophils and myeloperoxidase activity. Physiol Res 57:577–587
Gusman GS, Campana PR, Castro LC, Castilho RO, Teixeira MM, Braga FC 2015 Evaluation of the effects of some Brazilian medicinal plants on the production of TNF-alpha and CCL2 by THP-1 cells Evid Based Complement Alternat Med 2015 497123 https://doi.org/10.1155/2015/497123
Harborne JB, Greenham J, Williams CA, Eagles J, Markham KR (1993) Ten isoprenylated and C-methylated flavonoids from the leaves of three Vellozia species. Phytochemistry 34:219–226. https://doi.org/10.1016/S0031-9422(00)90808-2
Hawkins CL, Pattison DI, Davies MJ (2003) Hypochlorite-induced oxidation of amino acids, peptides and proteins. Amino Acids 25:259–274. https://doi.org/10.1007/s00726-003-0016-x
Jeong CH, Jeong HR, Choi GN, Kim DO, Lee U, Heo HJ (2011) Neuroprotective and anti-oxidant effects of caffeic acid isolated from Erigeron annuus leaf. Chin Med 6:25. https://doi.org/10.1186/1749-8546-6-25
Kato Y, Nagao A, Terao J, Osawa T (2003) Inhibition of myeloperoxidase-catalyzed tyrosylation by phenolic antioxidants in vitro. Biosci Biotechnol Biochem 67:1136–1139. https://doi.org/10.1271/bbb.67.1136
Klebanoff SJ (2005) Myeloperoxidase: friend and foe. J Leukoc Biol 77:598–625. https://doi.org/10.1189/jlb.1204697
Leite TC, de Sena AR, Dos Santos Silva TR, Dos Santos AK, Uetanabaro AP, Branco A (2012) Antimicrobial activity of Marcetia DC species (Melastomataceae) and analysis of its flavonoids by reverse phase-high performance liquid chromatography coupled-diode array detector. Pharmacogn Mag 8:209–214. https://doi.org/10.4103/0973-1296.99286
Mehrotra A, Shanbhag R, Chamallamudi MR, Singh VP, Mudgal J (2011) Ameliorative effect of caffeic acid against inflammatory pain in rodents. Eur J Pharmacol 666:80–86. https://doi.org/10.1016/j.ejphar.2011.05.039
Mykhailenko O, Gudzinskas Z, Kovalyov V, Desenko V, Ivanauskas L, Bezruk I, Georgiyants V (2020) Effect of ecological factors on the accumulation of phenolic compounds in Iris species from Latvia, Lithuania and Ukraine. Phytochem Anal 31:545–563. https://doi.org/10.1002/pca.2918
Queiroz RF, Vaz SM, Augusto O (2011) Inhibition of the chlorinating activity of myeloperoxidase by tempol: revisiting the kinetics and mechanisms. Biochem J 439:423–431. https://doi.org/10.1042/BJ20110555
Quintão FJO, Tavares RSN, Vieira-Filho SA, Souza GHB, Santos ODH (2013) Hydroalcoholic extracts of Vellozia squamata: study of its nanoemulsions for pharmaceutical or cosmetic applications. Rev Bras Farmacogn 23:101–107. https://doi.org/10.1590/S0102-695X2013005000001
Ribeiro B, Valentao P, Baptista P, Seabra RM, Andrade PB (2007) Phenolic compounds, organic acids profiles and antioxidative properties of beefsteak fungus (Fistulina hepatica). Food Chem Toxicol 45:1805–1813. https://doi.org/10.1016/j.fct.2007.03.015
Ruiz-Ojeda FJ, Olza J, Gil A, Aguilera CM. 2018 Oxidative stress and inflammation in obesity and metabolic syndrome In María AMMC García A (Eds.) Obesity Oxidative Stress and Dietary Antioxidants. Cambridge MA Academic Press pp
Sajjadi SE, Shokoohinia Y, Moayedi NS (2012) Isolation and identification of ferulic acid from aerial parts of Kelussia odoratissima Mozaff. Jundishapur J Nat Pharm Prod 7:159–162
Schrodinger L 2010 The PyMOL molecular graphics system (Version 1.3rl)
Septama AW, Jantan I, Panichayupakaranant P (2018) Flavonoids of Artocarpus heterophyllus Lam. heartwood inhibit the innate immune responses of human phagocytes. J Pharm Pharmacol 70:1242–1252. https://doi.org/10.1111/jphp.12952
Shiba Y, Kinoshita T, Chuman H, Taketani Y, Takeda E, Kato Y, Naito M, Kawabata K, Ishisaka A, Terao J, Kawai Y (2008) Flavonoids as substrates and inhibitors of myeloperoxidase: molecular actions of aglycone and metabolites. Chem Res Toxicol 21:1600–1609. https://doi.org/10.1021/tx8000835
Souza CD, Felfili JM (2006) The utilization of medicinal plants in the region of Alto Paraíso of Goiás, GO, Brazil. Acta Bot Bras 20:135–142. https://doi.org/10.1590/S0102-33062006000100013
Stierand K, Rarey M (2010) Drawing the PDB: protein−ligand complexes in two dimensions. ACS Med Chem Lett 1:540–545. https://doi.org/10.1021/ml100164p
Tian R, Ding Y, Peng YY, Lu N (2017) Inhibition of myeloperoxidase- and neutrophil-mediated hypochlorous acid formation in vitro and endothelial cell injury by (-)-epigallocatechin gallate. J Agric Food Chem 65:3198–3203. https://doi.org/10.1021/acs.jafc.7b00631
Trott O, Olson AJ (2010) AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem 31:455–461. https://doi.org/10.1002/jcc.21334
Tsumbu CN, Deby-Dupont G, Tits M, Angenot L, Frederich M, Kohnen S, Mouithys-Mickalad A, Serteyn D, Franck T (2012) Polyphenol content and modulatory activities of some tropical dietary plant extracts on the oxidant activities of neutrophils and myeloperoxidase. Int J Mol Sci 13:628–650. https://doi.org/10.3390/ijms13010628
Valentao P, Fernandes E, Carvalho F, Andrade PB, Seabra RM, Bastos ML (2003) Hydroxyl radical and hypochlorous acid scavenging activity of small centaury (Centaurium erythraea) infusion. A comparative study with green tea (Camellia sinensis). Phytomedicine 10:517–522. https://doi.org/10.1078/094471103322331485
Funding
This study was supported by grants from “Fundação de Amparo à Pesquisa do Estado da Bahia” (FAPESB RED038/2014; FAPESB RED012/2014) and “Conselho Nacional de Desenvolvimento Científico Tecnológico” (CNPq) (462401/2014–6).
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VMA performed the phytochemical analysis. IB and ERD contributed with the HPLC analysis. IJCV and RBF performed the NMR analyses. RFQ carried out the studies with MPO- and PMA-activated neutrophils. FHAL and BCS conducted the docking studies. VMA, ERD, and RFQ also performed the statistical analysis, interpreted the results, and drafted the manuscript. AB designed the studies and concluded the manuscript. All authors have read and approved the final manuscript.
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de Almeida, V.M., Dias, Ê.R., de Souza, B.C. et al. Myeloperoxidase Inhibition and In Silico Evaluation of Phenolics from Vellozia dasypus. Rev. Bras. Farmacogn. 33, 344–352 (2023). https://doi.org/10.1007/s43450-023-00375-w
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DOI: https://doi.org/10.1007/s43450-023-00375-w