Phytochemical Profiling and Biological Activities of Origanum onites L. Growing in Balıkesir, Türkiye

Abstract

Background and Aims: Origanum onites L., ordinarily known as Turkish oregano, have long been used in traditional medicine due to its therapeutic properties. This study aimed to investigate the phytochemical composition, antimicrobial activity, and cytotoxic effects of the essential oil and methanolic extract obtained from O. onites cultivated in the Balıkesir region of Türkiye. Methods: O. onites essential oil (OOEO) and methanolic extract were prepared through hydrodistillation and Soxhlet extraction, respectively. The antimicrobial potential was assessed using the broth microdilution method against a panel of bacterial and yeast strains. The cytotoxicity of A549 human lung adenocarcinoma cells was evaluated via 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The chemical profile of OOEO was determined by gas chromatography and mass spectrometry (GC-MS). Results: The essential oil exhibited antimicrobial activity with a minimum inhibitory concentration (MIC) of 120 μg/mL against Escherichia coli and Staphylococcus epidermidis, 240 μg/mL against Klebsiella pneumoniae, Staphylococcus aureus and Proteus mirabilis, 3900 μg/mL against Candida parapsilosis, and 7800 μg/mL against Candida tropicalis. Additionally, the cytotoxic effects were assessed on A549 cells, with Half-maximal inhibitory concentration (IC₅₀) values of 0.01 mg/mL for the OOEO and 0.03 mg/mL for the methanolic extract, indicating greater potency of the OOEO. Furthermore, oxidative stress markers suggested that both extracts could induce oxidative stress, as evidenced by the significant depletion of glutathione levels. GC-MS analysis of OOEO revealed a high concentration of bioactive constituents, particularly carvacrol (61.9%) and thymol (16.0%), both known for their potent biological effects. Conclusion: These findings highlight the potential of OOEO as an adjuvant therapy for lung cancer, under-scoring the need for further research to explore their mechanisms of action and therapeutic applications.

Keywords

• A549
• Antimicrobial
• Cytotoxicity
• Essential oil
• Origanum onites L.

References

1. Ali, H., Al-Khalifa, A. R., Aouf, A., Boukhebti, H., & Farouk, A. (2020). Effect of nanoen^ capsulation on volatile constituents, and antioxidant and anticancer activities of Algerian Origanum glandulosum Desf. essential oil. Scientific Reports, 10(1), 2812. https://doi.org/10.1038/s41598^020^59686^w google scholar
2. Altintas, A., Tabanca, N., Tyihák, E., Ott, P. G., Móricz, A. M., Mincsovics, E., & Wedge, D. E. (2013). Characterization of volatile constituents from Origanum onites and their antifungal and antibacterial activity. Journal of AOAC International, 96(6), 1200–1208. https://doi.org/10.5740/jaoacint.sgealtintas google scholar
3. Arafat, K., Sulaiman, S., Al^Azawi, A. M., Yasin, J., Sugathan, S., Nemmar, A., Karam S., & Attoub, S. (2022). Origanum majorana essential oil decreases lung tumor growth and metastasis in vitro and in vivo. Biomedicine & Pharmacotherapy, 155, Article 113762. https://doi.org/10.1016/j.biopha.2022.113762 google scholar
4. Ayaz, F., Köngül Şafak, E., Erkan Türkmen, K., Şeker Karatoprak, G., Katırcıoğlu, H., & Küçükboyacı, N. (2021). Assessment of antimicrobial, antibiofilm, and cyto^ toxic activities, and characterization of phenolic compounds of Origanum haussknechtii. Journal of Food Measurement and Characterization, 15(5), 4267– 4276. doi:10.1007/s11694^021^00984^w. google scholar
5. Baydar, H., Sağdiç, O., Özkan, G., & Karadoğan, T. Antibacterial activity and compo^ sition of essential oils from Origanum, Thymbra and Satureja species with commercial importance in Turkey. Food Control, 15(3), 169^172. https://doi.org/ 10.1016/S0956^7135(03)00028^8 google scholar
6. Bostancıoğlu, R. B., Kürkçüoğlu, M., Başer, K. H., & Koparal, A. T. (2012). Assessment of anti^angiogenic and anti^tumoral potentials of Origanum onites L. essential oil. Food and Chemical Toxicology: An International Journal Published for the British Industrial Biological Research Association, 50(6), 2002–2008. https:// doi.org/10.1016/j.fct.2012.03.074 google scholar
7. Canli, K., Bozyel, M. E., Turu, D., Benek, A., Simsek, O., & Altuner, E. M. (2023). Biochemical, antioxidant properties and antimicrobial activity of steno^en^ demic Origanum onites. Microorganisms, 11(8), 1987. https://doi.org/10.3390/ microorganisms11081987 google scholar
8. Cetin, A., Arslanbas, U., Saraymen, B., Canoz, O., Ozturk, A., & Sagdic, O. (2011). Efects of grape seed extract and Origanum onites essential oil on cisplatin^induced hepatotoxicity in rats. International Journal of Hematology and Oncology, 21, 133–140. https://doi.org/10.4999/uhod.10060 google scholar

9. Chauhan, A. K., & Kang, S. C. (2014). Thymol disrupts the membrane integrity of Salmonella ser. typhimurium in vitro and recovers infected macrophages from oxidative stress in an ex vivo model. Research in Microbiology, 165(7), 559–565. https://doi.org/10.1016/j.resmic.2014.07.001 google scholar
10. Chinou, I., Liolios, C., Moreau, D., & Roussakis, C. (2007). Cytotoxic activity of Origanum dictamnus. Fitoterapia, 78(5), 342–344. https://doi.org/10.1016/j.fitote.2007.02. 005 google scholar
11. Council of Europe. (2019). Oregano. European Pharmacopoeia. (p. 1565). 10th ed. Strasbourg: Council of Europe. google scholar
12. Emire, Z., & Yabalak, E. (2023). Can Origanum be a hope for cancer treatment? A review on the potential of Origanum species in preventing and treating cancers. International Journal of Environmental Health Research, 33(9), 894– 910. https://doi.org/10.1080/09603123.2022.2064437 google scholar
13. European Committe on Antimicrobial Susceptibility Testing (EUCAST). (2020). Break^ point tables for interpretation of MICs and zone diameters. Version 10.0. https://www.eucast.org/mic_and_zone_distributions_and_ecoffs google scholar
14. Ferahoğlu, E., Çürük, U., Çoğalan, D., Kırıcı, S., & Çakan, H. (2022). Determination of essential oil ratios and components of Origanum species grown in Çukurova conditions. Turkish Journal of Biodiversity, 5(2), 75^85. https://doi.org/10. 38059/biodiversity.1116233 google scholar
15. Guimarães, A. G., Oliveira, G. F., Melo, M. S., Cavalcanti, S. C., Antoniolli, A. R., Bonjardim, L. R., Silva, F. A., Santos, J. P., Rocha, R. F., Moreira, J. C., Araújo, A. A., Gelain, D. P., & Quintans^Júnior, L. J. (2010). Bioassay^guided evaluation of antioxidant and antinociceptive activities of carvacrol. Basic & Clinical Pharmacology & Toxicology, 107(6), 949–957. https://doi.org/10.1111/j.1742^7843.2010.00609.x google scholar
16. Karkabounas, S., Kostoula, O.K., Daskalou, T., Veltsistas, P., Karamouzis, M., Zelovitis, I., Metsios, A., Lekkas, P., Evangelou, A.M., Kotsis, N., & Skoufos, I. (2006). Anticar^ cinogenic and antiplatelet effects of carvacrol. Experimental Oncology, 28(2), 121^125. google scholar
17. Koparal, A. T., & Zeytinoglu, M. (2003). Effects of carvacrol on a human non^small cell lung cancer (NSCLC) cell line, A549. Cytotechnology, 43(1^3), 149–154. https:// doi.org/10.1023/b:cyto.0000039917.60348.45 google scholar
18. Kurkcuoglu, M., Agalar, H.G., Aksoy, A., & Baser, K.H.C. (2019). Composition of the essential oil of two endemic Helichrysum species in Turkey. Records of Natural Products, 13(3), 236^242. https://doi.org/10.25135/rnp.93.18.06.113 google scholar
19. Lagouri, V., Blekas, G., Tsimidou, M., Kokkini, S., & Boskou, D. (1993). Composition and antioxidant activity of essential oils from Oregano plants grown wild in Greece. Z. Lebensm. Unters Forsch, 197, 20–23. google scholar
20. Llana^Ruiz^Cabello, M., Gutiérrez^Praena, D., Puerto, M., Pichardo, S., Jos, Á., & Cameán, A. M. (2015). In vitro pro^oxidant/antioxidant role of carvacrol, thymol and their mixture in the intestinal Caco^2 cell line. Toxicology in vitro : An International Journal Published in Association with BIBRA, 29(4), 647–656. https://doi.org/ 10.1016/j.tiv.2015.02.006 google scholar
21. Llor, C., & Bjerrum, L. (2014). Antimicrobial resistance: risk associated with antibiotic overuse and initiatives to reduce the problem. Therapeutic Advances in Drug Safety, 5(6), 229–241. https://doi.org/10.1177/2042098614554919 google scholar
22. MAF; Ministry of Agriculture And Forestry, General Directorate of Agricultural Produc^ tion, Republic of Türkiye. (2020). Thyme Feasibility Report and Investor Guide, Türkiye. google scholar
23. Ozkan, G., Baydar, H., & Erbas, S. (2010). The influence of harvest time on essential oil composition, phenolic constituents and antioxidant properties of Turkish oregano (Origanum onites L.). Journal of the Science of Food and Agricul^ ture, 90(2), 205–209. https://doi.org/10.1002/jsfa.3788 google scholar
24. Piasecki, B., Balázs, V. L., Kieltyka^Dadasiewicz, A., Szabó, P., Kocsis, B., Horváth, G., & Ludwiczuk, A. (2023). Microbiological studies on the influence of essential oils from several origanum species on respiratory pathogens. Molecules, 28(7), 3044. https://doi.org/10.3390/molecules28073044 google scholar
25. Reis, R., Kolci, K., Yedikardes, E. N., Coskun, G. P., & Uzuner, Y. (2024). Dermal third^ hand smoke exposure induced epidermal alterations in human keratinocyte cells through oxidative damage and MMP^1 expression. Experimental Derma^ tology, 33(2), e15020. https://doi.org/10.1111/exd.15020 google scholar
26. Retnosari, R., Ali, A. H., Zainalabidin, S., Ugusman, A., Oka, N., & Latip, J. (2024). The recent discovery of a promising pharmacological scaffold derived from car^ vacrol: A review. Bioorganic & Medicinal Chemistry Letters, 109, 129826. https:// doi.org/10.1016/j.bmcl.2024.129826 google scholar
27. Seyhan, G., Akkaya, D., Kolci, K., Reis, R., Yazici, N., Nur Barut, E., & Barut, B. (2023). Insights into the biological activity and cytotoxic mechanism of Epimedium pubigerum. Chemistry & Biodiversity, 20(11), e202301003. https://doi.org/10. 1002/cbdv.202301003 google scholar
28. Sezen, E., Reis, R., Özhan, Y., Sipahi, H., Gimatdin, R., Gören, A. C., Kirmizibekmez, H. & Guzelmeric, E. (2025). Phytochemical characterization and evaluation of anti^inflammatory and anticancer activities of Cistus laurifolius originated propolis. European Food Research and Technology, 251(1), 151^164. https://doi. org/10.1007/s00217^024^04624^7 google scholar
29. Sharifi^Rad, M., Berkay Yılmaz, Y., Antika, G., Salehi, B., Tumer, T. B., Kulandaisamy Venil, C., Das, G., Patra, J. K., Karazhan, N., Akram, M., Iqbal, M., Imran, M., Sen, S., Acharya, K., Dey, A., & Sharifi^Rad, J. (2021). Phytochemical constituents, bio^ google scholar logical activities, and health^promoting effects of the genus Origanum. Phy^ totherapy Research: PTR, 35(1), 95–121. https://doi.org/10.1002/ptr.6785 google scholar
30. Shokrzadeh, M., Ahmadi, A., Chabra, A., Naghshvar, F., Salehi, F., Habibi, E., & Haghi^Aminjan, H. (2014). An ethanol extract of Origanum vulgare attenuates cyclophosphamide^induced pulmonary injury and oxidative lung damage in mice. Pharmaceutical Biology, 52(10), 1229–1236. https://doi.org/10.3109/ 13880209.2013.879908 google scholar
31. Sokmen, A., Abdel^Baki, A. A. S., Al^Malki, E. S., Al^Quraishy, S., & Abdel^Haleem, H. M. (2020). Constituents of essential oil of Origanum minutiflorum and its in vitro antioxidant, scolicidal and anticancer activities. Journal of King Saud University, 32(4), 2377^2382. https://doi.org/10.1016/j.jksus.2020.03.018 google scholar
32. Spyridopoulou, K., Fitsiou, E., Bouloukosta, E., Tiptiri^Kourpeti, A., Vamvakias, M., Ore^ opoulou, A., Papavassilopoulou, E., Pappa, A., & Chlichlia, K. (2019). Extraction, chemical composition, and anticancer potential of Origanum onites L. essen^ tial oil. Molecules, 24(14), 2612. https://doi.org/10.3390/molecules24142612 google scholar
33. Tepe, B., Cakir, A., & Sihoglu Tepe, A. (2016). Medicinal uses, phytochemistry, and pharmacology of Origanum onites (L.): A review. Chemistry & Biodiversity, 13(5), 504–520. https://doi.org/10.1002/cbdv.201500069 google scholar
34. Tomsuk, Ö., Kuete, V., Sivas, H., & Kürkçüoğlu, M. (2024). Effects of essential oil of Origanum onites and its major component carvacrol on the expression of toxicity pathway genes in HepG2 cells. BMC Complementary Medicine and Therapies, 24(1), 265. https://doi.org/10.1186/s12906^024^04571^6 google scholar
35. Toncer, O., Karaman, S., Kizil, S., & Diraz, E. Changes in essential oil composition of oregano (Origanum onites L.) due to diurnal variations at different develop^ ment stages. Notulae Botanicae Horti Agrobotanici Cluj^Napoca, 37(2), 177^181. https://doi.org/10.15835/nbha3723188. google scholar
36. Veljovic, K., Tesevic, V., Mitrovic, H., & Stankovic, M. (2023). Essential oil of Origanum minutiflorum exhibits anti^inflammatory and antioxidative effects in human bronchial cells and antimicrobial activity on lung pathogens. Journal of Herbal Medicine, 39, 100651. https://doi.org/10.1016/j.hermed.2023.100651 google scholar
37. Yokoyama, C., Sueyoshi, Y., Ema, M., Mori, Y., Takaishi, K., & Hisatomi, H. (2017). Induc^ tion of oxidative stress by anticancer drugs in the presence and absence of cells. Oncology Letters, 14(5), 6066–6070. https://doi.org/10.3892/ol.2017.6931 google scholar