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Publisher: Peytchinski Publishing Ltd.
ISSN: 1312-773X (Online)
Issue: 2025, vol. 31, issue4
Subject Area: Medicine
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DOI: 10.5272/jimab.2025314.6631
Published online: 01 December 2025
Original article
J of IMAB. 2025 Oct-Dec;31(4):6631-6635
IL-6 INDUCTION BY BETULINIC ACID AND ITS PROLINE ETHYL ESTER SALT
Lyuba Miteva1
, Antonia Grigorova1, Neli Dimitrova1, Paula Ossowicz-Rupniewska2, Maya Guncheva3,
1) Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora, Bulgaria.
2) Department of Organic Chemical Technology and Polymer Materials, Faculty of Chemical Technology and Engineering, West Pomeranian University of Technology, Szczecin, Poland.
3) Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria.
ABSTRACT:
Purpose: This study aims to compare the effects of betulinic acid (BA) and its proline ethyl ester ([ProOEt][BA]) on cell viability and IL-6 production by peripheral blood mononuclear cells (PBMCs).
Materials/Methods: PBMC were isolated from 10 mL of whole blood donated from healthy volunteers. PBMC were stimulated with 1 μg/mL Lipopolysaccharide (LPS), BA or [ProOEt][BA] at final concentrations of 2.5, 5, 10 and 20 μM. Cell viability was assessed using the MTT assay. IL-6 concentration in the culture supernatants was determined by enzyme-linked immunosorbent assay (ELISA) after 24 hours of incubation.
Results: Cell viability of PBMCs decreased in a dose-dependent manner upon stimulation with either BA or [ProOEt][BA]. A significant reduction in cell viability was observed with 10 and 20 μM concentrations of [ProOEt][BA] compared to BA after 24 hours of incubation. The lower concentrations of both compounds (2.5 and 5 μM) had comparable effects on cell viability. The estimated IC50 values after 24 hours of PBMC stimulation were approximately equal for both BA (8.42 μM) and [ProOEt][BA] (8.86 μM).
A dose-dependent effect on IL-6 production was also observed in PBMCs after in vitro stimulation with BA or [ProOEt][BA]. At the lowest concentration (2.5 μM), both BA and [ProOEt][BA] caused a more than five-fold decrease in IL-6 production compared to LPS-treated cells (523.25 pg/mL and 581.90 pg/mL vs. 2994.50 pg/mL, respectively).
Conclusion: [ProOEt][BA] demonstrated similar activity to BA on PBMCs. A dose-dependent reduction in PBMC viability and IL-6 production was observed after in vitro stimulation with either BA or [ProOEt][BA].
Keywords: Betulinic acid salts, peripheral blood mononuclear cells, cytokine,
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Please cite this article as: Miteva L, Grigorova A, Dimitrova N, Ossowicz-Rupniewska P, Guncheva M. IL-6 induction by betulinic acid and its proline ethyl ester salt. J of IMAB. 2025 Oct-Dec;31(4):6631-6635. [Crossref - 10.5272/jimab.2025314.6631]
Correspondence to: Lyuba Miteva, Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University; 11, Armeiska Str., 6000 Stara Zagora, Bulgaria; E-mail: lyuba.miteva@trakia-uni.bg
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Received: 11 July 2025
Published online: 01 December 2025