Edited by: Alejandro Urzua, University of Santiago, Chile
Reviewed by: Christian Agyare, Kwame Nkrumah University of Science and Technology, Ghana; Marina Sokovic, University of Belgrade, Serbia
*Correspondence: Magdalena Majdan,
This article was submitted to Ethnopharmacology, a section of the journal Frontiers in Pharmacology
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
The aim of the study was to characterize phytochemicals in an infusion of the aerial parts of tarragon
The popularity of spices as ingredients in food for the prevention and treatment of disease has increased in recent years. Tarragon (
Tarragon comes from eastern and central Europe, southern Russia, and western Asia (
The aim of this work was to evaluate an infusion of Russian tarragon as a potential food product for the prevention and treatment of inflammation and bacterial infections. We investigated the effects of infusion on proinflammatory functions of human neutrophils, such as ROS production and IL-8 and TNF-
Fetal bovine serum (FBS), f-MLP (N-Formylmethionyl-leucyl-phenylalanine), luminol, HEPES solution, RPMI 1640 medium, and L-glutamine were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Phosphate-buffered saline (PBS Ca2+-free) and penicillin-streptomycin were purchased from Gibco (Grand Island, USA). Lipopolysaccharide (LPS from
The aerial parts of Russian tarragon were collected in August 2014 from the experimental field of the Department of Vegetables and Medicinal Plants in Wilanów, Warsaw, Mazovian district, Poland (21°0099109 E 52°162209 N). The plant material was authenticated by Prof. Ewa Osińska (Warsaw University of Life Sciences, Poland) according to a guidebook (
A 3 g portion of air-dried plant material was poured into boiling water (250 mL), covered, and allowed to stand for 15 min (3×) in a tea infuser (Ambition, Warsaw, Poland). Extracts were then filtered and lyophilized (lyophilizer Telstar Cryodos 50, Telstar International, S.L., Terrassa, Spain), resulting in the following yields: sample 1 – 1.32 g, sample 2 – 0.98 g, sample 3 – 1.24 g. UHPLC-DAD-MS analysis was conducted using a Dionex Ultimate 3000RS system coupled with an Amazon SL ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany). The ion trap AmazonSL mass spectrometer was equipped with an ESI interface. The eluate was introduced into the ESI interface of the mass spectrometer without splitting. The parameters for the ESI source were as follows: nebulizer pressure 40 psi; dry gas flow 9 L/min; dry temperature 300°C; and capillary voltage 4.5 kV. Analysis was carried out using scanning from
The buffy coats were prepared from peripheral venous blood collected from healthy human donors (< 35 years old) at the Warsaw Blood Donation Centre. Donors were confirmed to be healthy and all tests carried out showed values within a normal range. Donors did not smoke or take any medications. The study conformed to the principles of the Declaration of Helsinki. Neutrophils were isolated using a standard method by dextran sedimentation and centrifugation in a Pancoll gradient (
Cytotoxicity was determined by flow cytometry using propidium iodide (PI) staining. After 24 h of incubation in the standard conditions (37°C, 5% CO2) with extracts or standards used as positive controls in tests, the neutrophils were harvested and centrifuged (1500 RPM; 10 min; 4°C), washed once with cold PBS, and re-suspended in 500 µL of PBS. Five microliters of PI (50 µg/mL) solution was added to the cell suspensions. After 15 min of incubation at room temperature, cells were analyzed by flow cytometry, and 10000 events were recorded per sample. Cells that displayed high permeability to PI were expressed as a percentage of PI (+) cells. Triton X was used as positive control.
The ROS production by f-MLP-stimulated neutrophils was determined using luminol-dependent chemiluminescence. ADI were tested at concentrations of 12.5, 25, 50, and 100 μg/mL. Following isolation, cells were suspended in 70 μL (Ca2+)-free HBSS. Cell suspension (3.0×105/mL) was incubated with 50 μL of the samples with tested concentrations of extract and 50 μL of luminol (100 μM). ROS production was initiated by the addition of 30 μL of f-MLP (0.1 μg/mL). Changes in the chemiluminescence were measured over a 40 min period at intervals of 2 min in a microplate reader (BioTek, Synergy 4) at 37°C. Background chemiluminescence produced by non-stimulated cells was also checked. The tested extract did not interfere with the chemiluminescence signal. As a positive control, quercetin was used at a concentration of 20 μM. The percentage of ROS production was calculated in comparison to the control without investigated tarragon water extract.
Neutrophils (2 × 106 cells/mL) were cultured in RPMI 1640 medium with 10% FBS, 10 mM HEPES, and 2 mM L-glutamine for 24 h at 37°C with 5% CO2 in the absence or presence of extract at final concentrations of 12.5, 25, 50, and 100 μg/mL (96-well plates, 1 mL per well) 1 h before stimulation LPS (100 ng/mL). After 24 h, plates were centrifuged (2000 RPM; 10 min; 4°C) and supernatants were collected. The release of cytokines by stimulated neutrophils was evaluated by enzyme-linked immunosorbent (ELISA) tests following the manufacturer’s instructions (BD Biosciences, San Jose, CA, USA or R&D Systems, Minneapolis, MN, USA). Dexamethasone at concentrations of 12.5, 25, and 50 μM and quercetin at concentration of 50 μM were used as a positive control for the release of IL-8 and TNF-
The results were expressed as the mean ± SEM of three independent experiments performed in triplicate. The statistical significance of differences between means and control was determined by ANOVA with Tukey’s
The antibacterial activity was assessed against Gram-positive (
Further, the comprehensive analysis of ADI was performed with the UHPLC-DAD-MS/MS method. Chromatograms (280 nm and 350 nm) of ADI are depicted in
UHPLC-DAD-ESI-MS/MS chromatograms of ADI recorded at 280 nm (black line) and 350 nm (red line).
Constituents annotated in ADI extract by UHPLC-DAD-ESI-MS.
Compounds | UV[nm] | Rt[min] | [M+H]+ (major and fragments |
[M-H]- (major and fragments |
Identification | References |
---|---|---|---|---|---|---|
215, 296, 325 | 11.7 | 355 (163) | 353 (191) | caffeoylquinic acid (I) | ( |
|
216, 278 | 12.0 | 477 (315, 153) | protocatechuic acid dihexoside | |||
212, 282,320 | 12.2 | 371 (209, 191) | caffeoyl hexaric acid | ( |
||
212, 260 | 13.4 | 359 (197) | syringic acid hexoside | ( |
||
215, 286, 323 | 14.0 | 355 (163) | 353 (191) | caffeoylquinic acid (II) | ( |
|
214 | 17.8 | 360 (163, 307), 325 (163,307) | 387 (179, 285, 341) | unknown | ||
217, 286, 324 | 20.9 | 355 (163) | 353 (191) | caffeoylquinic acid (III) | ( |
|
216, 286, 324 | 22.8 | 355 (163) | 353 (191, 179, 173) | caffeoylquinic acid (IV) | ( |
|
202 | 23.3 | 225 | 447 (401) | unknown | ||
235, 300 | 25.0 | 374 [M+Na]+ (195, 177) | 355 (193, 149) | ferulic acid hexoside | ( |
|
270, 333 | 30.2 | 595 (457, 577) | 593 (575, 503, 473, 383, 353) | vicenin - 2 apigenin 6,8-di-C-glucoside | ( |
|
217, 288, 324 | 34.1 | 544 (365) | 525, (481, 433, 301) | unknown | ||
207, 230, 319 | 34.9 | – | 355 (193, 149) | ferulic acid hexoside | ( |
|
215, 280, 342 | 37.8 | 465 | 463 (301) | isoquercitrin/hiperoside | ( |
|
215, 256, 355 | 39.8 | 611 (465, 303) | 609 (301, 487) | quercetin-3- |
( |
|
210, 260, 336 | 40.8 | 641 (495, 333), 333 (318, 222) | 639 (331) | patuletin rhamnosylhexoside | ( |
|
212, 262, 341 | 41.9 | 495 | 493 | patuletin hexoside | ( |
|
201, 260, 347 | 42.8 | 697 (535, 303) | 695 (651, 609, 301) | quercetin derivative | ( |
|
203, 255, 346 | 43.3 | 727 (565, 495, 333) | 725 (681, 639, 331) | patuletin 3- |
( |
|
216, 286, 325 | 45.0 | 517 (499, 355) | 515 (353, 191, 179,173) | di- |
( |
|
– | 45.9 | 625 (479, 317) | 623 (315) | isorhamnetin rhamnosylhexoside | ( |
|
216, 245, 292, 325 | 46.1 | 517 | 515 (353, 191, 179) | di- |
( |
|
– | 46.8 | 655 (509, 347) | 653 (345, 330) | syringetin 3- |
( |
|
216, 325 | 47.8 | 517 (499) | 515 (353, 191, 179, 173) | di- |
( |
|
210, 255, 344 | 48.7 | 741 (579, 509, 347) | 739 (695) | patuletin malonylrhamnosylhexoside | ( |
|
216, 293, 325 | 50.3 | 517 (499) | 515 (353, 191, 179, 173) | di- |
( |
|
220, 319 | 58.0 | – | 471 (402, 309, 240) | unknown | ||
216, 271, 312 | 65.5 | 581 (435, 419, 273) | 579 (271) | unknown | ||
– | 66.0 | 303 (285) | 301 (151) | quercetin | ( |
|
220, 285 | 66.9 | 287 (269) | 285 (267, 257, 241, 229) | 6-demethoxycapillarisin | ( |
|
216, 276, 315 | 68.1 | 259 | 257 (151) | davidigenin | ( |
|
220, 286 | 71.6 | 287 (241, 167, 147) | 285 (269, 263, 175, 163,151) | sakuranetin | ( |
|
196, 230, 263 | 72.3 | – | 299 (283, 191) | unknown | ||
211, 228, 297, 335 | 73.0 | – | 271 (256, 150), | 2′,4′-dihydroxy-4-methoxydihydrochalcone | ( |
Neutrophils, also known as polymorphonuclear cells (PMNs), after infiltration to the inflammation site, generate ROS. Stimulation by f-MLP (bacterial derived factor) results in degranulation and the significant release of ROS compared to the non-stimulated control (
Effect of the ADI at concentrations of 12.5-100 µg/mL on ROS production by f-MLP-stimulated neutrophils [%], mean ± SEM. Positive control quercetin (QU) at a concentration of 20 µM. Statistical significance of differences was established by ANOVA with
Effect of ADI at concentrations of 12.5 - 100 µg/mL on
The cytotoxic effect of ADI at concentrations of 12.5 - 100 µg/mL on LPS-stimulated neutrophils after 24 h incubation [%], mean ± SEM. Statistical significance of differences was established by ANOVA with
The antimicrobial activity of ADI was evaluated against nine strains of human pathogenic bacteria compared to the ampicillin (
Antimicrobial activity of ADI and ampicillin [mg/mL].
ADI[MIC] | Ampicylin[MIC] | |
---|---|---|
0.09 | <0.00003 | |
2.35 | 0.00024 | |
0.363 | 0.000025 | |
23.5 | 0.0001 | |
5.9 | 0.000025 | |
>94 | 0.0001 | |
47 | 0.00195 | |
94 | 0.0005 | |
11.75 | 0.0032 |
Various genetic and environmental factors may influence the biosynthesis and chemical compositions of plant secondary metabolites (
In the present study, the chemical composition of
The data are available on request to the corresponding author.
Investigation, Writing-Original Draft Preparation, Writing-Review and Editing: MM. Investigations: RH, SG, MC, AK, EO (plant material collection). Methodology, Review, and Editing: AK, MC, SG, RH.
This project was carried out with the use of CePT infrastructure financed by the European Union’s European Regional Development Fund within the Operational Program “Innovative economy” for 2007–2013. The access publication fees were financially supported by Medical University of Warsaw.
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.