<abstract xmlns="http://www.w3.org/1999/xhtml"><sec><h3>Aims</h3><p>Silymarin (SIL), a mixture of phenolic compounds, has a pleiotropic mode of action on various cell types, including immune cells. In this study, we investigated the concentration-dependent effect of SIL on proliferation of concanavalin A (CoA)-stimulated mouse spleen T lymphocytes, their viability, and secretion of IFN-g and IL-4 cytokines <italic>ex vivo</italic> in relation to gene expressions of transcription factors nuclear factor kappa B and Foxp3. In addition, metabolic activity of T cells was determined as changes in the mitochondrial membrane potential and apoptosis.</p></sec><sec><h3>Material/Methods</h3><p>Isolated splenocytes were stimulated with lectin CoA and treated with SIL atthe concentrations of 5, 10, 20, and 40 μg/ml for 70 h and unstimulated cells served as the control. Cultures of splenocytes were evaluated for proliferation index following BrdU incorporation and viability of cells after trypan blue staining. Gene expressions of transcription factors and cytokines were assessed using real-time PCR, whereas ELISA test was applied to measure cytokine secretion. Mitochondrial membrane potential and apoptosis were determined by flow cytometry.</p></sec><sec><h3>Results</h3><p>We demonstrated that CoA-activated mouse spleen T lymphocytes show different susceptibilities to low (£10 μg/ml) and higher (20 and 40 μg/ml) SIL concentrations. Low concentrations resulted in increased proliferation, cytokine secretion, and mitochondrial membrane potential and reduced transition of cells to apoptosis. High concentration of SIL had the opposite effect without exerting significant cytotoxicity and upregulated genes for cytokines and transcription factors on mRNA level. It is possible that individual subpopulations of T cells induced by CoA were differentially affected by the various SIL concentrations and the dose of 40 μg/ml had the profound suppressive effect. This correlated with the highest expression of Foxp3 factor, indicating that this dose stimulated preferential differentiation to Tregs lymphocytes.</p></sec><sec><h3>Conclusions</h3><p>Treatment with suitable doses of SIL can provide potential benefits in the modulation of host immune functions in various diseases.</p></sec></abstract>

AimsSilymarin (SIL), a mixture of phenolic compounds, has a pleiotropic mode of action on various cell types, including immune cells. In this study, we investigated the concentration-dependent effect of SIL on proliferation of concanavalin A (CoA)-stimulated mouse spleen T lymphocytes, their viability, and secretion of IFN-g and IL-4 cytokines ex vivo in relation to gene expressions of transcription factors nuclear factor kappa B and Foxp3. In addition, metabolic activity of T cells was determined as changes in the mitochondrial membrane potential and apoptosis.Material/MethodsIsolated splenocytes were stimulated with lectin CoA and treated with SIL atthe concentrations of 5, 10, 20, and 40 μg/ml for 70 h and unstimulated cells served as the control. Cultures of splenocytes were evaluated for proliferation index following BrdU incorporation and viability of cells after trypan blue staining. Gene expressions of transcription factors and cytokines were assessed using real-time PCR, whereas ELISA test was applied to measure cytokine secretion. Mitochondrial membrane potential and apoptosis were determined by flow cytometry.ResultsWe demonstrated that CoA-activated mouse spleen T lymphocytes show different susceptibilities to low (£10 μg/ml) and higher (20 and 40 μg/ml) SIL concentrations. Low concentrations resulted in increased proliferation, cytokine secretion, and mitochondrial membrane potential and reduced transition of cells to apoptosis. High concentration of SIL had the opposite effect without exerting significant cytotoxicity and upregulated genes for cytokines and transcription factors on mRNA level. It is possible that individual subpopulations of T cells induced by CoA were differentially affected by the various SIL concentrations and the dose of 40 μg/ml had the profound suppressive effect. This correlated with the highest expression of Foxp3 factor, indicating that this dose stimulated preferential differentiation to Tregs lymphocytes.ConclusionsTreatment with suitable doses of SIL can provide potential benefits in the modulation of host immune functions in various diseases.

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells <italic>in vitro</italic>

The University of Veterinary Medicine and Pharmacy

European Pharmaceutical Journal

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells 

AimsSilymarin (SIL), a mixture of phenolic compounds, has a pleiotropic mode of action on various cell types, including immune cells. In this study, we...

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells in vitro

Article Category: Original research article/Review

Published Online: Nov 18, 2020

Page range: 17 - 26

Received: Oct 02, 2019

Accepted: Dec 10, 2019

DOI: https://doi.org/10.2478/afpuc-2020-0003

Keywords
silymarin, mouse, splenocytes, proliferation, mitochondrial potential, apoptosis

© 2019 Hrčková G et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells in vitro

Article Category: Original research article/Review

Published Online: Nov 18, 2020

Page range: 17 - 26

Received: Oct 02, 2019

Accepted: Dec 10, 2019

DOI: https://doi.org/10.2478/afpuc-2020-0003

Keywordssilymarin, mouse, splenocytes, proliferation, mitochondrial potential, apoptosis

© 2019 Hrčková G et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
silymarin, mouse, splenocytes, proliferation, mitochondrial potential, apoptosis