The Effect of Virgin Olive Oil Phenolics Against Oxidative Mitochondrial and Nuclear DNA in Human Peripheral Blood Mononuclear Cells

Abstract

In recent years, evidence has accumulated that the beneficial effects of olive oil are related to not only its high oleic acid content, but also the antioxidant activity of its phenolic components. This study examined the effect of virgin olive oil phenolic extract (VOPE) on basal and oxidative stress-induced DNA damage within nuclear and mitochon-drial genomes of human peripheral blood mononuclear cells (PBMCs). Cells were isolated from the blood samples of the healthy volunteers, 35 year old two male and two female. Cells were pre-treated with VOPE at a dose range that is dietary-relevant and non-cytotoxic. To create oxidative stress, cells were exposed to hydrogen peroxide (H2O2, 200 µM) for 30 minutes. The DNA damages were measured by gene-specific QPCR method. Pretreatment with VOPE did not change the level of basal DNA damage in both mitochondrial DNA (mtDNA) and nuclear DNA (nDNA). H2O2-induced oxidative stress caused two-fold higher DNA damage in mtDNA (2.76 lesions/10kb) than nDNA (1.43 lesions/10kb). However, VOPE treatment before oxidative stress did not show any protective effects on both mtDNA and nDNA. As a result, two different phenolic extract concentrations that could be found in the tissues and vascular system after virgin olive oil consumption do not have a capacity to prevent oxidative DNA damage in human PBMCs. These results indicate that the beneficial effect of olive oil, which is known to have more on the digestive system, may be due to its higher concentrations immediately after consumption.

Keywords

• Phenolic compounds
• , gene-specific QPCR
• oxidative DNA damage
• oxidative stress

Sızma Zeytinyağı Fenoliklerinin İnsan Periferik Kan Mononükleer Hücrelerinde Oksidatif Mitokondriyal ve Nükleer DNA Hasarına Karşı Etkisi

Abstract

Son yıllarda, zeytinyağının faydalı etkilerinin sadece yüksek oleik asit içeriğiyle ilişkili değil, aynı zamanda içerdiği fenolik bileşenlerin antioksidan aktivitesiyle de ilişkili olduğu kanıtlanmıştır. Bu çalışmada, sızma zeytinyağı fenolik ekstraktının (ZFE), insan periferik kan mononükleer hücrelerinin (PKMH) nükleer ve mitokondriyal DNA'sındaki bazal DNA hasarı ve oksidatif stres kaynaklı DNA hasarı üzerindeki etkisi incelendi. Hücreler, 35 yaşındaki sağlıklı iki erkek ve iki kadın gönüllünün kan örneklerinden izole edildi. Hücrelere diyetle alınabilecek ve sitotoksik etkisi olmayan ZFE konsantrasyonları ile ön-uygulama yapıldı. Hücreler üzerinde oksidatif stres oluş-turmak için 30 dakika hidrojen peroksit (H2O2, 200 µM) ile uygulama yapıldı. DNA hasarları gene-özgü QPCR yöntemi ile ölçüldü. Hücrelere ZFE ile ön uygulama yapılması her iki genomdaki bazal DNA hasar seviyesi üzerin-de olumsuz bir etkiye neden olmadı. H2O2 ile oluşturulan oksidatif stres, nDNA (nükleer DNA) (1.43 hasar/10kb) ile karşılaştırıldığında mtDNA'da (mitokondriyal DNA) (2.76 hasar/10kb) iki kat daha fazla hasara neden oldu. Bununla birlikte, oksidatif stres öncesi ZFE ile ön-uygulama yapılması hem mtDNA hem de nDNA üzerinde koru-yucu etki göstermedi. Sonuçta, sızma zeytinyağı tüketimi sonrası doku ve vasküler sistemde bulunabilecek iki farklı fenolik ekstrakt konsantrasyonunun, insan PKMH’lerde oksidatif DNA hasarlarını önleyici bir etkiye sahip olmadığı belirlendi. Bu sonuçlar, zeytinyağının sindirim sistemi üzerinde daha fazla sahip olduğu bilinen faydalı etkisinin tüketiminden hemen sonraki daha yüksek konsantrasyonlarından kaynaklanabileceğini işaret etmektedir

Keywords

• Fenolik bileşikler
• gene-özgü QPCR
• oksidatif DNA hasarı
• oksidatif stres, zeytinyağı

References

1. Aebi, H. (1974). In catalase. In H.U. Bergmeyer (Ed.), In methods of enzymatic analysis (pp. 673–684). New York, USA. Erişim adresi: https://www.sciencedirect.com/book/9780120913022/methods-of-enzymatic-analysis
2. Bienert, G.P., Schjoerring, J.K. ve Jahn, T.P. (2006). Membrane transport of hydrogen peroxide, Biochimica et Biophysica Acta, 1758, 994–1003. https://doi.org/10.1016/j.bbamem.2006.02.015
3. Bishop, K.S., Erdrich, S., Karunasinghe, N., Han, D.Y., Zhu, S., Jesuthasan A. ve Ferguson L.R. (2015). An investigation into the association between DNA damage and dietary fatty acids in men with prostate cancer. Nutrients, 7, 405–422. https://doi.org/10.3390/nu7010405
4. Buckland, G. ve Gonzalez, C.A. (2015). The role of olive oil in disease prevention: a focus on the recent epidemiological evidence from cohort studies and dietary intervention trials. British Journal of Nutrition, 113, 94–101. https://doi.org/10.1017/S0007114514003936
5. Cline S.D. (2012). Mitochondrial DNA damage and its consequences for mitochondrial gene expression. Biochimica et Biophysica Acta, 1819(9-10), 979–991. https://doi.org/10.1016/j.bbagrm.2012.06.002
6. Devasagayam, T.P., Tilak, J.C., Boloor, K.K., Sane, K.S., Ghaskadbi, S.S. ve Lele, R.D. (2004). Free radicals and antioxidants in human health: current status and future prospects. The Journal of the Association of Physicians of India, 52, 794-804. https://doi.org/10.1016/j.bbagrm.2012.06.002
7. Erol, Ö., Arda, N. ve Erdem, G. (2012). Phenols of virgin olive oil protects nuclear DNA against oxidative damage in HeLa cells. Food and Chemical Toxicology, 50(10), 3475–3479. https://doi.org/10.1016/j.fct.2012.07.048
8. Fabiani, R., Rosignoli, P., De Bartolomeo, A., Fuccelli, R., Servili, M., Montedoro, G.F. ve Morozzi G. (2008). Oxidative DNA damage is prevented by extracts of olive oil, hydroxytyrosol, and other olive phenolic compounds in human blood mononuclear cells and HL60 cells. The Journal of Nutrition, 138(8), 1411–1416. https://doi.org/10.1093/jn/138.8.1411

9. Fitó, M., de la Torre, R., Farré-Albaladejo, M., Khymenetz, O., Marrugat, J. ve Covas, M.I. (2007). Bioavailability and antioxidant effects of olive oil phenolic compounds in humans: a review. Annali dell'Istituto Superiore di Sanità, 43(4), 375-381. http://old.iss.it/publ/anna/2007/4/434375.pdf
10. Furda, A., Santos, J.H., Meyer, J.N. ve van Houten, B. (2014). Quantitative PCR-based measurement of nuclear and mitochondrial DNA damage and repair in mammalian cells. In P. Keohavong ve S. Grant (Ed.), Molecular toxicology protocols. Methods in molecular biology (Methods and protocols) (pp. 419-437). Totowa, NJ. https://doi.org/10.1007/978-1-62703-739-6_31
11. Gill, PK. (2019). Rapid isolation of peripheral blood mononuclear cells from whole blood with ficoll hypaque density centrifugation. Journal of International Research in Medical and Pharmaceutical Sciences, 14(1), 17-20. https://www.ikprress.org/index.php/JIRMEPS/article/view/4566
12. Giovannelli, L,. Saieva, C., Masala, G., Testa, G., Salvini, S., Pitozzi, V., Riboli, E., Dolara, P. ve Palli, D. (2002). Nutritional and lifestyle determinants of DNA oxidative damage: a study in a mediterranean population. Carcinogenesis, 23(9), 1483-1489. https://doi.org/10.1093/carcin/23.9.1483
13. Gorzynik-Debicka, M., Przychodzen, P., Cappello, F., Kuban-Jankowska, A., Marino Gammazza, A., Knap, N., Wozniak, M. ve Gorska-Ponikowska, M. (2018). Potential health benefits of olive oil and plant polyphenols. International Journal of Molecular Sciences, 28: 19(3). https://doi.org/10.3390/ijms19030686
14. Hunter, S.E., Jung, D., Di Giulio, R.T. ve Meyer, J.N. (2010). The QPCR assay for analysis of mitochondrial DNA damage, repair, and relative copy number. Methods, 51(4), 444–451. https://doi.org/10.1016/j.ymeth.2010.01.033
15. Jang, Y.C. ve van Remmen, H. (2009). The mitochondrial theory of aging: insight from transgenic and knockout mouse models. Experimental Gerontology, 44(4), 256–260. https://doi.org/10.1016/j.exger.2008.12.006
16. Konstantinidou, V., Covas, M.I., Sola, R. ve Fitó, M. (2013). Up-to date knowledge on the in vivo transcriptomic effect of the mediterranean diet in humans. Molecular Nutrition Food Research, 57(5), 772–783. https://doi.org/10.1002/mnfr.201200613
17. Kryston, T.B., Georgiev, A.B., Pissis, P. ve Georgakilas, A.G. (2011). Role of oxidative stress and DNA damage in human carcinogenesis. Mutation Research, 711(1-2), 193-201. https://doi.org/10.1016/j.mrfmmm.2010.12.016
18. Martín-Peláez, S., Covas, M.I., Fitó, M., Kušar, A. ve Pravst, I. (2013). Health effects of olive oil polyphenols: recent advances and possibilities for the use of health claims. Molecular Nutrition Food Research, 57(5), 760–771. https://doi.org/10.1002/mnfr.201200421
19. Montedoro, G., Servili, M., Baldioli, M. ve Miniati, E. (1992). Simple and hydrolyzable phenolic compounds in virgin olive oil. 1. Their extraction, separation, and quantitative and semiquantitative evaluation by HPLC. The Journal of Agricultural and Food Chemistry, 40, 1571–1576. https://doi.org/10.1021/jf00021a019
20. Nousis, L., Doulias, P.T., Aligiannis, N., Bazios, D., Agalias, A., Galaris, D. ve Mitakou, S. (2005). DNA protecting and genotoxic effects of olive oil related components in cells exposed to hydrogen peroxide. Free Radical Research, 39(7), 787–795. https://doi.org/10.1080/10715760500045806
21. Palli, D., Vineis, P., Russo, A., Berrino, F., Krogh, V., Masala, G., Munnia, A., Panico, S., Taioli, E., Tumino, R., Garte, S. ve Peluso, M. (2000). Diet, metabolic polymorphisms and DNA adducts: the epıc-İtaly cross-sectional study. The International Journal of Cancer, 87, 444–451. https://doi.org/10.1002/1097-0215(20000801)87:3<444::AID-IJC21>3.0.CO;2-%23
22. Ragazzi, E. ve Veronese, G. (1973). Quantitative analysis of phenolic compounds after thin-layer chromatographic separation, Journal of Chromatography, 77(2), 369–375. https://doi.org/10.1016/S0021-9673(00)92204-0
23. Romani, A., Ieri, F., Urciuoli, S., Noce, A., Marrone, G., Nediani, C. ve Bernini, R. (2019). Health effects of phenolic compounds found in extra-virgin olive oil, by-products, and leaf of Olea europaea L. Nutrients, 11(8). pii: E1776. https://doi.org/10.3390/nu11081776
24. Rubió, L., Valls, R.M., MacIà, A., Pedre, A., Giralt, M., Romero, M.P., De La Torre, R., Covas, M.I., Solà, R. ve Motilva, M.J. (2012). Impact of olive oil phenolic concentration on human plasmatic phenolic metabolites. Food Chemistry, 135(4), 2922-2929. https://doi.org/10.1016/j.foodchem.2012.07.085
25. Salvini, S., Sera, F., Caruso, D., Giovannell, L., Visioli, F., Saieva, C., Masala, G., Ceroti, M., Giovacchini, V., Pitozzi, V., Galli, C., Romani, A., Mulinacci, N., Bortolomeazzi, R., Dolara, P. ve Palli D. (2006). Daily consumption of a high-phenol extra-virgin olive oil reduces oxidative DNA damage in postmenopausal women. British Journal of Nutrition, 95(4), 742–751. https://doi.org/10.1079/BJN20051674
26. Weinbrenner, T., Fitó, M., de la Torre, R., Saez, G.T., Rijken, P., Tormos, C., Coolen, S., Albaladejo, M.F., Abanades, S., Schroder, H., Marrugat, J. ve Covas, M.I. (2004). Olive oils high in phenolic compounds modulate oxidative/antioxidative status in men. The Journal of Nutrition, 134(9), 2314-2321. https://doi.org/10.1093/jn/134.9.2314