Protective Effects of Nitrated Fatty Acids from Olive Oil on Cardiovascular Risk in Non-Alcoholic Fatty Liver Disease

Authors

DOI:

https://doi.org/10.31285/AGRO.30.1717

Keywords:

cellular respiration, heart, liver, nitroalkenes, olive oil

Abstract

Non-alcoholic fatty liver disease (NAFLD) is defined by the deposition of fat in the liver without alcohol consumption. Moreover, it is the most prevalent chronic liver disease worldwide and is associated with an increased risk of cardiovascular disease. Olive oil is one of the main sources of lipids in the Mediterranean diet and may reduce cardiovascular risk, although the mechanism has not yet been elucidated. Olive oil is a source of nitrated fatty acids (NO₂-FA) that are formed during digestion and are bioactive molecules with potent anti-inflammatory actions and cytoprotective effects. The aim of this work was to study the capacity of NO₂-FA derived from olive oil in protecting mitochondrial dysfunction in cardiomyocytes in NAFLD. We used a murine model of NAFLD induced by a high-fat diet, supplemented with and without nitrite, as well as with and without olive oil. Heart biopsies were used to analyze mitochondrial function using high-resolution respirometry and the enzymatic activity of the mitochondrial transport chain complexes. As a result, we observed that the supplementation of olive oil in the presence of nitrite, a condition that favors the formation of NO₂-FA, improves mitochondrial function in heart biopsies, which correlates with the improvement in ATPase activity in that tissue. These compounds may be partly responsible for the beneficial effects of this functional food.

Downloads

Download data is not yet available.

References

Abenavoli, L., Milanović, M., Milić, N., Luzza, F., & Giuffrè, A. M. (2019). Olive oil antioxidants and non-alcoholic fatty liver disease. Expert Review of Gastroenterology & Hepatology, 13(8), 739-749. https://doi.org/10.1080/17474124.2019.1634544

Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. https://doi.org/10.1016/0003-2697(76)90527-3

Brand, M. D., & Nicholls, D. G. (2011). Assessing mitochondrial dysfunction in cells. The Biochemical Journal, 435(2), 297-312. https://doi.org/10.1042/BJ20110162

Buchan, G. J., Bonacci, G., Fazzari, M., Salvatore, S. R., & Gelhaus Wendell, S. (2018). Nitro-fatty acid formation and metabolism. Nitric Oxide, 79, 38-44. https://doi.org/10.1016/j.niox.2018.07.003

Bullón-Vela, V., Abete, I., Tur, J. A., Pintó, X., Corbella, E., Martínez-González, M. A., Toledo, E., Corella, D., Macías, M., Tinahones, F., Fitó, M., Estruch, R., Ros, E., Salas-Salvadó, J., Daimiel, L., Zulet, M. A., Martínez, J. A., & PREDIMED Plus investigators (2020). Influence of lifestyle factors and staple foods from the Mediterranean diet on non-alcoholic fatty liver disease among older individuals with metabolic syndrome features. Nutrition, 71, Article 110620. https://doi.org/10.1016/j.nut.2019.110620

Bulotta, S., Celano, M., Lepore, S. M., Montalcini, T., Pujia, A., & Russo, D. (2014). Beneficial effects of the olive oil phenolic components oleuropein and hydroxytyrosol: Focus on protection against cardiovascular and metabolic diseases. Journal of Translational Medicine, 12, Article 219. https://doi.org/10.1186/s12967-014-0219-9

Byrne, C. D., & Targher, G. (2015). NAFLD: A multisystem disease. Journal of Hepatology, 62(1 Suppl), S47-S64. https://doi.org/10.1016/j.jhep.2014.12.012

Cassina, A., & Radi, R. (1996). Differential inhibitory action of nitric oxide and peroxynitrite on mitochondrial electron transport. Archives of Biochemistry and Biophysics, 328(2), 309-316. https://doi.org/10.1006/abbi.1996.0178

Castro, L., Demicheli, V., Tórtora, V., & Radi, R. (2011). Mitochondrial protein tyrosine nitration. Free Radical Research, 45(1), 37-52. https://doi.org/10.3109/10715762.2010.516254

Caussy, C., Aubin, A., & Loomba, R. (2021). The relationship between type 2 diabetes, NAFLD, and cardiovascular risk. Current Diabetes Reports, 21(5), Article 15. https://doi.org/10.1007/s11892-021-01383-7

Day, C. P., & James, O. F. (1998). Steatohepatitis: A tale of two "hits"? Gastroenterology, 114(4), 842-845. https://doi.org/10.1016/s0016-5085(98)70599-2

Deprince, A., Haas, J. T., & Staels, B. (2020). Dysregulated lipid metabolism links NAFLD to cardiovascular disease. Molecular Metabolism, 42, Article 101092. https://doi.org/10.1016/j.molmet.2020.101092

Ditano-Vázquez, P., Torres-Peña, J. D., Galeano-Valle, F., Pérez-Caballero, A. I., Demelo-Rodríguez, P., Lopez-Miranda, J., Katsiki, N., Delgado-Lista, J., & Alvarez-Sala-Walther, L. A. (2019). The fluid aspect of the Mediterranean diet in the prevention and management of cardiovascular disease and diabetes: The role of polyphenol content in moderate consumption of wine and olive oil. Nutrients, 11(11), Article 2833. https://doi.org/10.3390/nu11112833

Duell, P. B., Welty, F. K., Miller, M., Chait, A., Hammond, G., Ahmad, Z., Cohen, D. E., Horton, J. D., Pressman, G. S., Toth, P. P., American Heart Association Council on Arteriosclerosis, Thrombosis and Vascular Biology; Council on Hypertension; Council on the Kidney in Cardiovascular Disease; Council on Lifestyle and Cardiometabolic Health; & Council on Peripheral Vascular Disease. (2022). Nonalcoholic fatty liver disease and cardiovascular risk: A scientific statement from the American Heart Association. Arteriosclerosis, Thrombosis, and Vascular Biology, 42(6), e168-e185. https://doi.org/10.1161/ATV.0000000000000153

Fazzari, M., Khoo, N. K., Woodcock, S. R., Jorkasky, D. K., Li, L., Schopfer, F. J., & Freeman, B. A. (2017). Nitro-fatty acid pharmacokinetics in the adipose tissue compartment. Journal of Lipid Research, 58(2), 375-385. https://doi.org/10.1194/jlr.M072058

Fazzari, M., Trostchansky, A., Schopfer, F. J., Salvatore, S. R., Sánchez-Calvo, B., Vitturi, D., Valderrama, R., Barroso, J. B., Radi, R., Freeman, B. A., & Rubbo, H. (2014). Olives and olive oil are sources of electrophilic fatty acid nitroalkenes. PloS One, 9(1), Article e84884. https://doi.org/10.1371/journal.pone.0084884

Gaforio, J. J., Visioli, F., Alarcón-de-la-Lastra, C., Castañer, O., Delgado-Rodríguez, M., Fitó, M., Hernández, A. F., Huertas, J. R., Martínez-González, M. A., Menendez, J. A., Osada, J., Papadaki, A., Parrón, T., Pereira, J. E., Rosillo, M. A., Sánchez-Quesada, C., Schwingshackl, L., Toledo, E., & Tsatsakis, A. M. (2019). Virgin olive oil and health: Summary of the III International Conference on Virgin Olive Oil and Health Consensus Report, JAEN (Spain) 2018. Nutrients, 11(9), Article 2039. https://doi.org/10.3390/nu11092039

García-Roche, M., Casal, A., Carriquiry, M., Radi, R., Quijano, C., & Cassina, A. (2018). Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies. Redox Biology, 17, 207-212. https://doi.org/10.1016/j.redox.2018.03.008

García-Ruiz, I., Fernández-Moreira, D., Solís-Muñoz, P., Rodríguez-Juan, C., Díaz-Sanjuán, T., Muñoz-Yagüe, T., & Solís-Herruzo, J. A. (2010). Mitochondrial complex I subunits are decreased in murine nonalcoholic fatty liver disease: Implication of peroxynitrite. Journal of Proteome Research, 9(5), 2450-2459. https://doi.org/10.1021/pr9011427

García-Ruiz, I., Solís-Muñoz, P., Fernández-Moreira, D., Muñoz-Yagüe, T., & Solís-Herruzo, J. A. (2015). In vitro treatment of HepG2 cells with saturated fatty acids reproduces mitochondrial dysfunction found in nonalcoholic steatohepatitis. Disease Models & Mechanisms, 8(2), 183-191. https://doi.org/10.1242/dmm.018234

Geisler, A. C., & Rudolph, T. K. (2012). Nitroalkylation--a redox sensitive signaling pathway. Biochimica et Biophysica Acta, 1820(6), 777-784. https://doi.org/10.1016/j.bbagen.2011.06.014

Gorzynik-Debicka, M., Przychodzen, P., Cappello, F., Kuban-Jankowska, A., Marino Gammazza, A., Knap, N., Wozniak, M., & Gorska-Ponikowska, M. (2018). Potential health benefits of olive oil and plant polyphenols. International Journal of Molecular Sciences, 19(3), Article 686. https://doi.org/10.3390/ijms19030686

Guasch-Ferré, M., Hu, F. B., Martínez-González, M. A., Fitó, M., Bulló, M., Estruch, R., Ros, E., Corella, D., Recondo, J., Gómez-Gracia, E., Fiol, M., Lapetra, J., Serra-Majem, L., Muñoz, M. A., Pintó, X., Lamuela-Raventós, R. M., Basora, J., Buil-Cosiales, P., Sorlí, J. V., Ruiz-Gutiérrez, V., … & Salas-Salvadó, J. (2014). Olive oil intake and risk of cardiovascular disease and mortality in the PREDIMED Study. BMC Medicine, 12, Article 78. https://doi.org/10.1186/1741-7015-12-78

Hill, B. G., Benavides, G. A., Lancaster, J. R., Jr, Ballinger, S., Dell'Italia, L., Jianhua, Z., & Darley-Usmar, V. M. (2012). Integration of cellular bioenergetics with mitochondrial quality control and autophagy. Biological Chemistry, 393(12), 1485-1512. https://doi.org/10.1515/hsz-2012-0198

Kakimoto, P. A., & Kowaltowski, A. J. (2016). Effects of high fat diets on rodent liver bioenergetics and oxidative imbalance. Redox Biology, 8, 216-225. https://doi.org/10.1016/j.redox.2016.01.009

Kansanen, E., Bonacci, G., Schopfer, F. J., Kuosmanen, S. M., Tong, K. I., Leinonen, H., Woodcock, S. R., Yamamoto, M., Carlberg, C., Ylä-Herttuala, S., Freeman, B. A., & Levonen, A. L. (2011). Electrophilic nitro-fatty acids activate NRF2 by a KEAP1 cysteine 151-independent mechanism. The Journal of Biological Chemistry, 286(16), 14019-14027. https://doi.org/10.1074/jbc.M110.190710

Kansanen, E., Jyrkkänen, H. K., Volger, O. L., Leinonen, H., Kivelä, A. M., Häkkinen, S. K., Woodcock, S. R., Schopfer, F. J., Horrevoets, A. J., Ylä-Herttuala, S., Freeman, B. A., & Levonen, A. L. (2009). Nrf2-dependent and -independent responses to nitro-fatty acids in human endothelial cells: Identification of heat shock response as the major pathway activated by nitro-oleic acid. The Journal of Biological Chemistry, 284(48), 33233-33241. https://doi.org/10.1074/jbc.M109.064873

Kasper, P., Martin, A., Lang, S., Kütting, F., Goeser, T., Demir, M., & Steffen, H. M. (2021). NAFLD and cardiovascular diseases: A clinical review. Clinical Research in Cardiology, 110(7), 921-937. https://doi.org/10.1007/s00392-020-01709-7

Khoo, N. K. H., & Schopfer, F. J. (2019). Nitrated fatty acids: from diet to disease. Current Opinion in Physiology, 9, 67-72. https://doi.org/10.1016/j.cophys.2019.04.013

Khoo, N. K. H., Fazzari, M., Chartoumpekis, D. V., Li, L., Guimaraes, D. A., Arteel, G. E., Shiva, S., & Freeman, B. A. (2019). Electrophilic nitro-oleic acid reverses obesity-induced hepatic steatosis. Redox Biology, 22, Article 101132. https://doi.org/10.1016/j.redox.2019.101132

Kien, C. L., Bunn, J. Y., & Ugrasbul, F. (2005). Increasing dietary palmitic acid decreases fat oxidation and daily energy expenditure. The American Journal of Clinical Nutrition, 82(2), 320-326. https://doi.org/10.1093/ajcn.82.2.320

Koenitzer, J. R., Bonacci, G., Woodcock, S. R., Chen, C. S., Cantu-Medellin, N., Kelley, E. E., & Schopfer, F. J. (2016). Fatty acid nitroalkenes induce resistance to ischemic cardiac injury by modulating mitochondrial respiration at complex II. Redox Biology, 8, 1-10. https://doi.org/10.1016/j.redox.2015.11.002

Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259), 680-685. https://doi.org/10.1038/227680a0

Mancebo-Campos, V., Salvador, M. D., & Fregapane, G. (2023). EFSA health claims-based virgin olive oil shelf-life. Antioxidants (Basel), 12(8), Article 1563. https://doi.org/10.3390/antiox12081563

Mazzoli, A., Crescenzo, R., Cigliano, L., Spagnuolo, M. S., Cancelliere, R., Gatto, C., & Iossa, S. (2019). Early hepatic oxidative stress and mitochondrial changes following western diet in middle aged rats. Nutrients, 11(11), Article 2670. https://doi.org/10.3390/nu11112670

Pesta, D., & Gnaiger, E. (2012). High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods in Molecular Biology, 810, 25-58. https://doi.org/10.1007/978-1-61779-382-0_3

Pullman, M. E., Penefsky, H. S., Datta, A., & Racker, E. (1960). Partial resolution of the enzymes catalyzing oxidative phosphorylation: I. Purification and properties of soluble dinitrophenol-stimulated adenosine triphosphatase. The Journal of Biological Chemistry, 235, 3322-3329.

Reboredo-Rodríguez, P., Figueiredo-González, M., González-Barreiro, C., Simal-Gándara, J., Salvador, M. D., Cancho-Grande, B., & Fregapane, G. (2017). State of the art on functional virgin olive oils enriched with bioactive compounds and their properties. International Journal of Molecular Sciences, 18(3), Article 668. https://doi.org/10.3390/ijms18030668

Rudolph, V., Rudolph, T. K., Schopfer, F. J., Bonacci, G., Woodcock, S. R., Cole, M. P., Baker, P. R., Ramani, R., & Freeman, B. A. (2010). Endogenous generation and protective effects of nitro-fatty acids in a murine model of focal cardiac ischaemia and reperfusion. Cardiovascular Research, 85(1), 155-166. https://doi.org/10.1093/cvr/cvp275

Salabei, J. K., Gibb, A. A., & Hill, B. G. (2014). Comprehensive measurement of respiratory activity in permeabilized cells using extracellular flux analysis. Nature Protocols, 9(2), 421-438. https://doi.org/10.1038/nprot.2014.018

Sánchez-Calvo, B., Cassina, A., Mastrogiovanni, M., Santos, M., Trias, E., Kelley, E. E., Rubbo, H., & Trostchansky, A. (2021). Olive oil-derived nitro-fatty acids: Protection of mitochondrial function in non-alcoholic fatty liver disease. The Journal of Nutritional Biochemistry, 94, Article 108646. https://doi.org/10.1016/j.jnutbio.2021.108646

Sánchez-Calvo, B., Cassina, A., Rios, N., Peluffo, G., Boggia, J., Radi, R., Rubbo, H., & Trostchansky, A. (2016). Nitro-arachidonic acid prevents angiotensin II-induced mitochondrial dysfunction in a cell line of kidney proximal tubular cells. PloS One, 11(3), Article e0150459. https://doi.org/10.1371/journal.pone.0150459

Santos, R. D., Valenti, L., & Romeo, S. (2019). Does nonalcoholic fatty liver disease cause cardiovascular disease?: Current knowledge and gaps. Atherosclerosis, 282, 110-120. https://doi.org/10.1016/j.atherosclerosis.2019.01.029

Schopfer, F. J., & Khoo, N. K. H. (2019). Nitro-fatty acid logistics: Formation, biodistribution, signaling, and pharmacology. Trends in Endocrinology and Metabolism, 30(8), 505-519. https://doi.org/10.1016/j.tem.2019.04.009

Schopfer, F. J., Vitturi, D. A., Jorkasky, D. K., & Freeman, B. A. (2018). Nitro-fatty acids: New drug candidates for chronic inflammatory and fibrotic diseases. Nitric Oxide, 79, 31-37. https://doi.org/10.1016/j.niox.2018.06.006

Sharma, R. S., Harrison, D. J., Kisielewski, D., Cassidy, D. M., McNeilly, A. D., Gallagher, J. R., Walsh, S. V., Honda, T., McCrimmon, R. J., Dinkova-Kostova, A. T., Ashford, M. L. J., Dillon, J. F., & Hayes, J. D. (2017). Experimental nonalcoholic steatohepatitis and liver fibrosis are ameliorated by pharmacologic activation of Nrf2 (NF-E2 p45-Related Factor 2). Cellular and Molecular Gastroenterology and Hepatology, 5(3), 367-398. https://doi.org/10.1016/j.jcmgh.2017.11.016

Singer, T. P. (1974). Determination of the activity of succinate, NADH, choline, and alpha-glycerophosphate dehydrogenases. Methods of Biochemical Analysis, 22, 123-175. https://doi.org/10.1002/9780470110423.ch3

Takeuchi, H., Matsuo, T., Tokuyama, K., Shimomura, Y., & Suzuki, M. (1995). Diet-induced thermogenesis is lower in rats fed a lard diet than in those fed a high oleic acid safflower oil diet, a safflower oil diet or a linseed oil diet. The Journal of Nutrition, 125(4), 920-925. https://doi.org/10.1093/jn/125.4.920

Targher, G., Byrne, C. D., & Tilg, H. (2024). MASLD: A systemic metabolic disorder with cardiovascular and malignant complications. Gut, 73(4), 691-702. https://doi.org/10.1136/gutjnl-2023-330595

Terés, S., Barceló-Coblijn, G., Benet, M., Alvarez, R., Bressani, R., Halver, J. E., & Escribá, P. V. (2008). Oleic acid content is responsible for the reduction in blood pressure induced by olive oil. Proceedings of the National Academy of Sciences of the United States of America, 105(37), 13811-13816. https://doi.org/10.1073/pnas.0807500105

Valenzuela, R., Espinosa, A., Llanos, P., Hernandez-Rodas, M. C., Barrera, C., Vergara, D., Romero, N., Pérez, F., Ruz, M., & Videla, L. A. (2016). Anti-steatotic effects of an n-3 LCPUFA and extra virgin olive oil mixture in the liver of mice subjected to high-fat diet. Food & Function, 7(1), 140-150. https://doi.org/10.1039/c5fo01086a

Visioli, F., Franco, M., Toledo, E., Luchsinger, J., Willett, W. C., Hu, F. B., & Martinez-Gonzalez, M. A. (2018). Olive oil and prevention of chronic diseases: Summary of an International conference. Nutrition, Metabolism, and Cardiovascular Diseases, 28(7), 649-656. https://doi.org/10.1016/j.numecd.2018.04.004

Wright, M. M., Kim, J., Hock, T. D., Leitinger, N., Freeman, B. A., & Agarwal, A. (2009). Human haem oxygenase-1 induction by nitro-linoleic acid is mediated by cAMP, AP-1 and E-box response element interactions. The Biochemical Journal, 422(2), 353-361. https://doi.org/10.1042/BJ20090339

Downloads

Published

2026-06-05

How to Cite

Sánchez-Calvo, B., Cassina, A., Rubbo, H., & Trostchansky, A. (2026). Protective Effects of Nitrated Fatty Acids from Olive Oil on Cardiovascular Risk in Non-Alcoholic Fatty Liver Disease. Agrociencia Uruguay, 30(NE2), e1717. https://doi.org/10.31285/AGRO.30.1717

Issue

Section

Health and Consumption Section

Altmetric

Article metrics
Abstract views
Galley views
PDF Views
HTML views
Other views
Scan to share
QR code to open and share this publication

Most read articles by the same author(s)