Effect of the secretome of multipotent mesenchymal stromal cells induced by dexamethasone in vitro on the expression of phospho-NF-κB p65 and Ki-67 in mononuclear cells

Authors

DOI:

https://doi.org/10.3897/rrpharmacology.7.68533

Abstract

Introduction: To investigate the influence of secretomes from native and dexamethasone-treated adipose-derived multipotent mesenchymal stromal cells (MMSC) on the proliferation of mononuclear cells (MNCs) and on their expression of phospho-NF-κB p65 in vitro.

Materials and Methods: MMSCs were isolated from the fat of 5 healthy donors. The cells were grown in culture up to passage four, then treated with dexamethasone for 3 hours, washed off the preparations, and incubated in a serum-free medium for 48 hours. Some of the cells were not treated with dexamethasone. Supernatants from cell cultures were concentrated by ultrafiltration, standardized by the content of galectin-1, sterilized, and added to MNCs from peripheral blood of 8 healthy donors. MNCs were isolated in a Ficoll density gradient according to a standard protocol. The expression of phospho-NF-κB p65 and Ki-67 in MNCs under the influence of MMSC secretomes in isotypic and negative controls was determined on a flow cytometer.

Results and Discussion: The expression of phospho-NF-kκB p65 and Ki-67 is decreased by the MMSC secretome. At the same time, a statistically significant decrease in phospho-NF-κB p65 by 36.2% (p< 0.05) is observed when using a secretome from native cells. Ki-67 expression is reduced by 42.3% (p < 0.05) under the influence of a secretome from dexamethasone-treated MMSCs.

Conclusion: The MMSC secretome, as well as MMSCs themselves, has an anti-inflammatory effect due to the effect on the expression of the active form of NF-κB and the proliferative activity of mononuclear cells. At the same time, pretreatment of cells with dexamethasone reduces the effect on phospho-NF-κB expression and increases the inhibitory effect on MNC proliferation.

Keywords:

multipotent mesenchymal stromal cells, dexamethasone, mononuclear cells, inflammation, NF-κB, RELA, p65, Ki-67

Author Contribution

Polina A. Golubinskaya, Biruch Center of Cell Technologies LLC

Cell engineer.

Maksim V. Puzanov, Biruch Center of Cell Technologies LLC

CEO.

Svetlana Yu. Burda, Biruch Center of Cell Technologies LLC

Co-investigator.

Darya A. Kostina, Research Institute of Live Systems Pharmacology

Teaching assistant, Department of Pharmacology and Clinical Pharmacology.

Yuriy E. Burda, Biruch Center of Cell Technologies LLC

PhD in Medicine, R&D Director.

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Published

30-06-2021

How to Cite

Golubinskaya PA, Puzanov MV, Burda SY, Kostina DA, Burda YE (2021) Effect of the secretome of multipotent mesenchymal stromal cells induced by dexamethasone in vitro on the expression of phospho-NF-κB p65 and Ki-67 in mononuclear cells. Research Results in Pharmacology 7(2): 101–107. https://doi.org/10.3897/rrpharmacology.7.68533

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Section

Review article

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