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Gentiana lutea root aqueous extract mitigates hydroxyurea-induced genotoxicity through antioxidative action and DNA repair: an in vitro study in healthy human peripheral blood mononuclear cells Cover

Gentiana lutea root aqueous extract mitigates hydroxyurea-induced genotoxicity through antioxidative action and DNA repair: an in vitro study in healthy human peripheral blood mononuclear cells

Open Access
|Dec 2025

Figures & Tables

Figure 1

Radical-scavenging capacity of GRE determined with the DPPH (A) and ABTS assays (B), expressed as a percentage of inhibition. GRE – yellow gentian (Gentiana lutea L.) root extract; DPPH – 2,2-diphenyl-1-picrylhydrazyl; ABTS – 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid
Radical-scavenging capacity of GRE determined with the DPPH (A) and ABTS assays (B), expressed as a percentage of inhibition. GRE – yellow gentian (Gentiana lutea L.) root extract; DPPH – 2,2-diphenyl-1-picrylhydrazyl; ABTS – 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid

Figure 2

Genotoxicity (MN frequency, A and C panels) and cytotoxicity (CBPI, B and D panels) in PBMC cultures treated with GRE or HU alone. *P<0.05, **P<0.01, ***P<0.001 treatments vs control. CBPI – cytokinesis-block proliferation index; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea; MN – micronucleus; PBMC – peripheral blood mononuclear cell
Genotoxicity (MN frequency, A and C panels) and cytotoxicity (CBPI, B and D panels) in PBMC cultures treated with GRE or HU alone. *P<0.05, **P<0.01, ***P<0.001 treatments vs control. CBPI – cytokinesis-block proliferation index; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea; MN – micronucleus; PBMC – peripheral blood mononuclear cell

Figure 3

Representative photomicrographs of binuclear cells (CBMN assay): HU treatment (A, B, C), micronuclei (red arrows) and nucleoplasmic bridges (blue arrows); GRE pre-treatment and HU treatment (D, E, F), micronuclei (red arrows) and nuclear buds (green arrows) (scale bar=10 µm). CBMN – cytokinesis-block micronucleus; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea
Representative photomicrographs of binuclear cells (CBMN assay): HU treatment (A, B, C), micronuclei (red arrows) and nucleoplasmic bridges (blue arrows); GRE pre-treatment and HU treatment (D, E, F), micronuclei (red arrows) and nuclear buds (green arrows) (scale bar=10 µm). CBMN – cytokinesis-block micronucleus; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea

Figure 4

Genoprotective potential of GRE (CBMN assay): MN frequency (A) and CBPI (B) in PBMC cultures pre-treated with GRE followed by HU treatment. #P<0.05; ##P<0.01; ###P<0.001 – GRE pre-treatment vs HU treatment; ***P<0.001 – HU treatment vs control. CBPI – cytokinesis-block proliferation index; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea; MN – micronucleus; PBMC – peripheral blood mononuclear cell
Genoprotective potential of GRE (CBMN assay): MN frequency (A) and CBPI (B) in PBMC cultures pre-treated with GRE followed by HU treatment. #P<0.05; ##P<0.01; ###P<0.001 – GRE pre-treatment vs HU treatment; ***P<0.001 – HU treatment vs control. CBPI – cytokinesis-block proliferation index; GRE – yellow gentian (Gentiana lutea L.) root extract; HU – hydroxyurea; MN – micronucleus; PBMC – peripheral blood mononuclear cell

Figure 5

Oxidative stress parameters (A and B panels) and gene expression (C panel) in PBMC in cultures exposed to GRE and HU. #P<0.05 – GRE pre-treatment vs HU treatment; **P<0.01 and ***P<0.001 – GRE pre-treatment vs control. GRE – yellow gentian (Gentiana lutea L.) root extract; GSH – reduced glutathione; HU – hydroxyurea; PAB – pro-oxidant/antioxidant balance; PBMC – peripheral blood mononuclear cell
Oxidative stress parameters (A and B panels) and gene expression (C panel) in PBMC in cultures exposed to GRE and HU. #P<0.05 – GRE pre-treatment vs HU treatment; **P<0.01 and ***P<0.001 – GRE pre-treatment vs control. GRE – yellow gentian (Gentiana lutea L.) root extract; GSH – reduced glutathione; HU – hydroxyurea; PAB – pro-oxidant/antioxidant balance; PBMC – peripheral blood mononuclear cell

Frequencies of micronuclei, nuclear buds, and nucleoplasmic bridges in individual treatments (optimisation experiment) and PBMC cultures pre-treated with GRE followed by HU treatment

Micronucleus frequency (per 1000 binuclear cells)Nuclear bud frequency (per 1000 binuclear cells)Nucleoplasmic bridge frequency (per 1000 binuclear cells)
Optimisation experiment
Control8.17±0.46.33±0.49/
HU 50 µmol/L13.07±0.412±0.891.66±0.31
HU 100 µmol/L18.67±0.3317.83±0.953.17±0.31
HU 200 µmol/L20.25±0.6318.67±1.284.67±0.49
GRE 0.25 mg/mL5.75±0.448.83±0.6/
GRE 0.5 mg/mL4.67±0.548.5±0.76/
GRE 1 mg/mL5.67±0.49.5±0.92/
GRE 2 mg/mL8.25±0.610.83±0.83/
Pre-treatment (100 µmol/L HU+GRE)
Control7.83±0.75.5±0.43/
HU 100 µmol/L16.83±0.4816.17±0.87/
GRE 0.25 mg/mL+HU11.92±1.1925.67±0.84/
GRE 0.5 mg/mL+HU10.58±0.825.17±0.65/
GRE 1 mg/mL+HU11.33±0.6735.67±1.98/
GRE 2 mg/mL+HU9.25±0.9330.17±1.2/
DOI: https://doi.org/10.2478/aiht-2025-76-4033 | Journal eISSN: 1848-6312 | Journal ISSN: 0004-1254
Language: English, Croatian, Slovenian
Page range: 282 - 291
Submitted on: Aug 1, 2025
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Accepted on: Nov 1, 2025
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Published on: Dec 30, 2025
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2025 Ksenija Radošević, Mila Kostić, Marijana Janić, Ivan Jovanović, Maja Živković, Ana Valenta Šobot, Jelena Filipović Tričković, published by Institute for Medical Research and Occupational Health
This work is licensed under the Creative Commons Attribution 4.0 License.