journal article·2026-06-23Open access

The Addition of Propolis and Royal Jelly to Chestnut and Thyme Honey Reduces DNA Damage Induced by Food Mutagens in HepG2 Cells by the Comet Assay

Amaia Iriondo-DeHond, Ana I. Haza, Vanesa Sánchez-Martín, Paloma Morales

Applied Sciences· Volume 16· Issue 13

Abstract

N-nitrosamines and acrylamide are food mutagens classified as “probably carcinogenic to humans (Group 2A)” by the International Agency for Research on Cancer (IARC) from evidence of carcinogenicity. One of the main objectives of food safety is to reduce the presence of these substances in food. Therefore, the present study aimed to evaluate the effect of the addition of propolis, royal jelly or a combination of both bee products (2–10%) to chestnut and thyme honey on their protective properties against food mutagen-induced genotoxicity. DNA damage was evaluated by the alkaline comet assay. N-nitrosamines (N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR)) and acrylamide (AA) induced genotoxicity in human hepatoma HepG2 cells. All tested samples at all concentrations used (0.1–10 µg/mL) decreased genotoxic effects of the three food mutagens. The protective effects of honey samples and mixtures towards DNA damage induced by food mutagens were in the following order: NDMA > AA > NPYR, being more effective against NDMA compared to AA and NPYR. The mixtures of chestnut honey with 10% propolis, or 10% royal jelly, and 10% propolis showed a greater protective effect against NDMA, NPYR and AA compared to the honey sample alone. This protective activity may be attributable to the phenolic compound content and antioxidant capacity exhibited by the analyzed samples. Overall, the results suggest that thyme and chestnut honey supplemented with bee-derived products could represent potential natural chemopreventive candidates against food-borne mutagens.
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Research context

OpenAlex topics: Bee Products Chemical Analysis · Potato Plant Research · Mycotoxins in Agriculture and Food

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Sección Departamental de Nutrición y Ciencia de los Alimentos, Departamento de Nutrición y Ciencia de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain

Funding

Ministerio de Ciencia e Innovación

References (39)

Show source-provided references
2. Govindaraju, I., Sana, M., Chakraborty, I., Rahman, M.H., Biswas, R., and Mazumder, N. (2024). Dietary Acrylamide: A Detailed Review on Formation, Detection, Mitigation, and Its Health Impacts. Foods, 13.10.3390/foods13040556 ↗
3. (2023, May 14). European Commission Standard IARC Classification. Available online: https://ec.europa.eu/health/scientific_committees/opinions_layman/en/electromagnetic-fields/glossary/ghi/iarc-classification.htm#top.
4. European Food Safety Authority (2015). Scientific Opinion on acrylamide in food. EFSA J., 13, 4104.10.2903/j.efsa.2015.4104 ↗
5. European Food Safety Authority (EFSA) (2024, May 07). Nitrosamines in Food Raise a Health Concern. Available online: https://www.efsa.europa.eu/en/news/nitrosamines-food-raise-health-concern.
6. Antioxidant activity and reduction potential of dietary herbs on acrylamide formation in asparagine-glucose reaction model10.1080/10942912.2023.2244701 ↗
7. Therapeutic Implications of Dietary Polyphenols-Loaded Nanoemulsions in Cancer Therapy10.1021/acsabm.3c01205 ↗
8. Dietary polyphenols in chemoprevention and synergistic effect in cancer: Clinical evidences and molecular mechanisms of action10.1016/j.phymed.2021.153554 ↗
9. Moskwa, J., Naliwajko, S.K., Dobiecka, D., and Socha, K. (2023). Bee products and colorectal cancer—Active components and mechanism of action. Nutrients, 15.10.3390/nu15071614 ↗
10. Martinello, M., and Mutinelli, F. (2021). Antioxidant activity in bee products: A review. Antioxidants, 10.10.3390/antiox10010071 ↗
11. Sawicki, T., Starowicz, M., Kłębukowska, L., and Hanus, P. (2022). The Profile of Polyphenolic Compounds, Contents of Total Phenolics and Flavonoids, and Antioxidant and Antimicrobial Properties of Bee Products. Molecules, 27.10.3390/molecules27041301 ↗
12. Ministerio de Agricultura Pesca y Alimentación (2021). El Sector Apícola en 2020: Principales Magnitudes e Indicadores Economicos.
13. Assessment of sensory properties and in vitro antimicrobial activity of monofloral Sicilian honey10.1016/j.fbio.2023.102449 ↗
14. Sánchez-Martín, V., Haza, A.I., Iriondo-DeHond, A., Del Castillo, M.D., Hospital, X.F., Fernández, M., Hierro, E., and Morales, P. (2022). Protective effect of thyme and chestnut honeys enriched with bee products against benzo(a)pyrene-induced DNA damage. Int. J. Environ. Res. Public Health, 19.10.3390/ijerph192416969 ↗
15. Sánchez-Martín, V., Morales, P., González-Porto, A.V., Iriondo-DeHond, A., López-Parra, M.B., Del Castillo, M.D., Hospital, X.F., Fernández, M., Hierro, E., and Haza, A.I. (2022). Enhancement of the antioxidant capacity of thyme and chestnut honey by addition of bee products. Foods, 11.10.3390/foods11193118 ↗
16. Assessment of the predictive capacity of the optimized in vitro comet assay using HepG2 cells10.1016/j.mrgentox.2018.01.010 ↗
17. Protocols in in vitro hepatocyte research
18. Factors influencing DNA migration from individual cells subjected to gel electrophoresis10.1016/0014-4827(92)90378-L ↗
19. Minimum information for reporting on the comet assay (MIRCA): Recommendations for describing comet assay procedures and results10.1038/s41596-020-0398-1 ↗
20. Protective effects of isothiocyanates towards N- nitrosamine-induced DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay10.1002/jat.1163 ↗
21. Rosemary, heather and heterofloral honeys protect towards cytotoxicity of acrylamide in human hepatoma cells
22. The Protective Effect of Whole Honey and Phenolic Extract on Oxidative DNA Damage in Mice Lymphocytes Using Comet Assay10.1007/s11130-017-0634-1 ↗
23. Assessment of phenolic profile of Turkish honeys10.1080/10942912.2016.1188307 ↗
24. The Ameliorative Effect of Royal Jelly against Malathion Genotoxicity in Bone Marrow and Liver of Rat
25. Royal jelly reduce DNA damage induced by alkylating agent in mice
26. Phenolic profile, antioxidant capacities and enzymatic inhibitory activities of propolis from different geographical areas: Needs for analytical harmonization
27. Protective Effect of Honey and Propolis against Gentamicin-Induced Oxidative Stress and Hepatorenal Damages10.1155/2021/9719906 ↗
28. Evaluation of antigenotoxic effects of plant flavonoids quercetin and rutin on HepG2 cells10.1002/ptr.3436 ↗
29. Propolis alleviates 4-aminobiphenyl-induced oxidative DNA damage by inhibition of CYP2E1 expression in human liver cells10.1002/tox.23147 ↗
30. Assessment of the radioprotective effect of propolis in breast cancer patients undergoing radiotherapy. New perspective for an old honey bee product
31. Habryka, C., Socha, R., and Juszczak, L. (2020). The effect of enriching honey with propolis on the antioxidant activity, sensory characteristics, and quality parameters. Molecules, 25.10.3390/molecules25051176 ↗
32. Camacho-Bernal, G.I., Cruz-Cansino, N.D.S., Ramírez-Moreno, E., Delgado-Olivares, L., Zafra-Rojas, Q.Y., Castañeda-Ovando, A., and Suárez-Jacobo, Á. (2021). Addition of bee products in diverse food sources: Functional and physicochemical properties. Appl. Sci., 11.10.3390/app11178156 ↗
33. Protective effects of isothiocyanates alone or in combination with vitamin C towards N -nitrosodibutylamine or N -nitrosopiperidine-induced oxidative DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay10.1002/jat.1270 ↗
34. Chaudhry, G.E.S., Md Akim, A., Sung, Y.Y., and Sifzizul, T.M.T. (2022). Cancer and apoptosis: The apoptotic activity of plant and marine natural products and their potential as targeted cancer therapeutics. Front. Pharmacol., 13.10.3389/fphar.2022.842376 ↗
35. Inhibition of CYP2E1 with natural agents may be a feasible strategy for minimizing the hepatotoxicity of ethanol10.1054/mehy.1999.1015 ↗
36. Inhibition of cytochrome P450 by propolis in human liver microsomes10.5487/TR.2016.32.3.207 ↗
37. Bioactive Components and Antioxidant and Antibacterial Activities of Different Varieties of Honey: A Screening Prior to Clinical Application10.1021/acs.jafc.8b05436 ↗
38. Protective effect of vitamin C towards N -nitrosamine-induced DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay10.1016/j.tiv.2007.03.015 ↗
39. Organic honey supplementation reverses pesticide-induced genotoxicity by modulating DNA damage response10.1002/mnfr.201600005 ↗