Evaluation of estrogenic activity of red clover (Trifolium pratense L.) sprouts cultivated under different conditions by content of isoflavones, calorimetric study and molecular modelling

Budryn, Grażyna; Gałązka-Czarnecka, Ilona; Brzozowska, Ewelina; Grzelczyk, Joanna; Mostowski, Radosław; Żyżelewicz, Dorota; Cerón‐Carrasco, José P.; Pérez‐Sánchez, Horacio

doi:10.1016/j.foodchem.2017.10.100

Cited by 34 publications

(47 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Red clover seeds (Trifolium pratense L.) were obtained from FN Granum (Wodzierady, Poland). Seed germination and fermentation carried out with Lactobacillus casei 0979 strain were performed as described previously [10,12]. Lactic acid fermentation was performed to increase the concentration of phenolics and to convert some of the glycosides into aglycones.…”

Section: Preparation Of Red Clover Extracts and Phenolic Compounds Dementioning

confidence: 99%

“…For biological activity assays, preparations were dissolved in a phosphate buffer solution (PBS)/dimethyl sulfoxide (DMSO) (1:1 v/v) solution at a concentration of 300 mg of extract/mL. The contents of isoflavones were determined as described previously by Budryn et al [12] via LC-ESI-MS analysis of isoflavones with the use of a Shimadzu (Kyoto, Japan) liquid chromatograph equipped with a SPD M20A diode array detector and quadrupole m mass spectrometer 2020 with an ion source of electrospray type (ESI) in positive scan mode with Kinetex C18 column after elution and MS identification. The content of isoflavones was calculated on dry mass and presented as concentration of particular isoflavones, as well as total free aglycones and total isoflavones ( Table 1).…”

Section: Preparation Of Red Clover Extracts and Phenolic Compounds Dementioning

confidence: 99%

“…The phenolic composition of red clover (Trifolium pratense L.) sprouts was described in detail previously in [10,12,13]. The isoflavone content in CU, CUH, CUF and CUFH extracts was determined Nutrients 2020, 12, 257 8 of 26 by the LC-ESI-MS method and is summarized in Table 1.…”

Section: Isoflavone Compounds Profile and Contentmentioning

confidence: 99%

“…Our previous results of isothermal titration calorimetry (ITC) modelling and docking simulation with red clover sprouts' isoflavones clearly showed their higher affinity to ERβ than ERα, indicating their great potential to advantageously activate ERβ without leading to breast cancer cells' proliferation [12]. What is more, further comparable studies revealed red clover main isoflavones' ability to form spontaneous bond with Lys 18 located in ATP binding site of actin through formation of π-π-type linkages [13].…”

mentioning

confidence: 91%

“…Red clover displays high contents of isoflavone aglycons-biochanin A and formononetin, which are 4 -O-methylated plant precursors to genistein and daidzein. These methoxylated aglycones are quickly absorbed into the bloodstream from the gastrointestinal tract without the need for their hydrolysis with bacterial glycosidases, in contrast to glycosides [10].Our previous results of isothermal titration calorimetry (ITC) modelling and docking simulation with red clover sprouts' isoflavones clearly showed their higher affinity to ERβ than ERα, indicating their great potential to advantageously activate ERβ without leading to breast cancer cells' proliferation [12]. What is more, further comparable studies revealed red clover main isoflavones' ability to form spontaneous bond with Lys 18 located in ATP binding site of actin through formation of π-π-type linkages [13].…”

mentioning

confidence: 99%

See 4 more Smart Citations

The Effects of Trifolium pratense L. Sprouts’ Phenolic Compounds on Cell Growth and Migration of MDA-MB-231, MCF-7 and HUVEC Cells

Zakłos‐Szyda

Budryn

2020

Nutrients

Self Cite

View full text Add to dashboard Cite

Uncontrolled growth and migration and invasion abilities are common for cancer cells in malignant tumors with low therapeutic effectiveness and high mortality and morbidity. Estrogen receptor β (ERβ), as a member of the nuclear receptor superfamily, shows potent tumor suppressive activities in many cancers. Phytoestrogens’ structural resemblance to 17 β-estradiol allows their binding to ERβ isoform predominantly, and therefore, expression of genes connected with elevated proliferation, motility and invasiveness of cancer cells may be downregulated. Among polyphenolic compounds with phytoestrogenic activity, there are isoflavones from Trifolium pratense L. (red clover) sprouts, containing high amounts of formononetin and biochanin A and their glycosides. To determine the source of the most biologically active isoflavones, we obtained four extracts from sprouts before and after their lactic fermentation and/or β-glucosidase treatment. Our previous results of ITC (isothermal titration calorimetry) modelling and a docking simulation showed clover isoflavones’ affinity to ERβ binding, which may downregulate cancer cell proliferation and migration. Thus, the biological activity of T. pratense sprouts’ extracts was checked under in vitro conditions against highly invasive human breast cancer cell line MDA-MB-231 and non-invasive human breast cancer cell line MCF-7 cells. To compare extracts’ activities acquired for cancer cells with those activities against normal cells, as a third model we choose human umbilical vein endothelial cells (HUVEC), which, due to their migration abilities, are involved in blood vessel formation. Extracts obtained from fermented sprouts at IC0 dosages were able to inhibit migration of breast cancer cells through their influence on intracellular ROS generation; membrane stiffening; adhesion; regulation of MMP-9, N-cadherin and E-cadherin at transcriptional level; or VEGF secretion. Simultaneously, isolated phenolics revealed no toxicity against normal HUVEC cells. In the manuscript, we proposed a preliminary mechanism accounting for the in vitro activity of Trifolium pratense L. isoflavones. In this manner, T. pratense sprouts, especially after their lactic fermentation, can be considered a potent source of biological active phytoestrogens and a dietary supplement with anti-cancer and anti-invasion properties.

show abstract

Section: Preparation Of Red Clover Extracts and Phenolic Compounds Dementioning

confidence: 99%

Section: Preparation Of Red Clover Extracts and Phenolic Compounds Dementioning

confidence: 99%