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(-)-Epigallocatechin-3-gallate and hydroxytyrosol improved antioxidative and anti-inflammatory responses in bovine mammary epithelial cells

  • L. Basiricò (a1), P. Morera (a1), D. Dipasquale (a1), R. Bernini (a1), L. Santi (a1), A. Romani (a2), N. Lacetera (a1) and U. Bernabucci (a1)...

Abstract

(-)-Epigallocatechin-3-gallate (EGCG), the major phenolic compound of green tea, and hydroxytyrosol (HTyr), a phenol found in olive oil, have received attention due to their wide-ranging health benefits. To date, there are no studies that report their effect in bovine mammary gland. Therefore, the aim of this study was to evaluate the anti-oxidative and anti-inflammatory effects of EGCG and HTyr in bovine mammary epithelial cell line (BME-UV1) and to compare their antioxidant and anti-inflammatory in vitro efficacy. Sample of EGCG was obtained from a commercially available green tea extract while pure HTyr was synthetized in our laboratories. The mammary oxidative stress and inflammatory responses were assessed by measuring the oxidative stress biomarkers and the gene expression of inflammatory cytokines. To evaluate the cellular antioxidant response, glutathione (GSH/GSSH), γ-glutamylcysteine ligase activity, reactive oxygen species and malondialdehyde (MDA) production were measured after 48-h incubation of 50 µM EGCG or 50 µM of HTyr. Reactive oxygen species production after 3 h of hydrogen peroxide (50 µM H2O2) or lipopolysaccharide (20 µM LPS) exposure was quantified to evaluate and to compare the potential protection of EGCG and HTyr against H2O2-induced oxidative stress and LPS-induced inflammation. The anti-inflammatory activity of EGCG and HTyr was investigated by the evaluation of pro and anti-inflammatory interleukins (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and IL-10) messenger RNA abundance after treatment of cells for 3 h with 20 µM of LPS. Data were analyzed by one-way ANOVA. (-)-Epigallocatechin-3-gallate or HTyr treatments induced higher concentrations of intracellular GSH compared to control cells, matched by an increase of γ-glutamylcysteine ligase activity mainly in cells treated with HTyr. Interestingly, EGCG and HTyr prevented oxidative lipid damage in the BME-UV1 cells by a reduction of intracellular MDA levels. (-)-Epigallocatechin-3-gallate and HTyr were able to enhance cell resistance against H2O2-induced oxidative stress. It was found that EGCG and HTyr elicited a reduction of the three inflammatory cytokines TNF-α, IL-1β, IL-6 and an increase of the anti-inflammatory cytokine IL-10. Hydroxytyrosol has proved to be a strong antioxidant compound, and EGCG has shown mainly an anti-inflammatory profile. These results indicated that EGCG and HTyr may provide dual protection because they were able to attenuate oxidative stress and inflammatory responses, suggesting that these phenolic compounds are potential natural alternatives to be used in dairy cattle as feed supplement for reducing the development of oxidative and inflammatory processes related to parturition or as topical treatments for the control of bovine intramammary inflammation.

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Corresponding author

E-mail: bernab@unitus.it

References

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Abuelo, A, Hernández, J, Benedito, JL and Castillo, C 2015. The importance of the oxidative status of dairy cattle in the periparturient period: revisiting antioxidant supplementation. Journal of Animal Physiology and Animal Nutrition (Berl) 99, 10031016. doi: 10.1111/jpn.12273.
Aitken, SL, Karcher, EL, Rezamand, P, Gandy, JC, VandeHaar, MJ, Capuco, AV and Sordillo, LM 2009. Evaluation of antioxidant and proinflammatory gene expression in bovine mammary tissue during the periparturient period. Journal of Dairy Science 92, 589598. doi: 10.3168/jds.2008-1551.
Arango Duque, G and Descoteaux, A 2014. Macrophage cytokines: involvement in immunity and infectious diseases. Frontiers in Immunology 5, 112. doi: 10.3389/fimmu.2014.00491.
Arévalo Turrubiarte, M, Perruchot, MH, Finot, L, Mayeur, F and Dessauge, F 2016. Phenotypic and functional characterization of two bovine mammary epithelial cell lines in 2D and 3D models. American Journal of Physiology-Cell Physiology 310, 348356. doi: 10.1152/ajpcell.00261.2015.
Basiricò, L, Morera, P, Dipasquale, D, Tröscher, A, Serra, A, Mele, M and Bernabucci, U 2015. Conjugated linoleic acid isomers strongly improve the redox status of bovine mammary epithelial cells (BME-UV1). Journal of Dairy Science 98, 70717082. doi: 10.3168/jds.2015-9787.
Basiricò, L, Morera, P, Dipasquale, D, Tröscher, A, and Bernabucci, U 2017. Comparison between CLA and essential fatty acids in preventing oxidative stress in bovine mammary epithelial cells (BME-UV1). Journal of Dairy Science 100, 22992309. doi: 10.3168/jds.2016-11729.
Bernabucci, U, Ronchi, B, Lacetera, N and Nardone, A 2005. Influence of body condition score on relationships between metabolic status and oxidative stress in periparturient dairy cows. Journal of Dairy Science 88, 20172026. doi: 10.3168/jds.S0022-0302( 05)72878-2.
Bernini, R, Gilardini Montani, MS, Merendino, N, Romani, A and Velotti, F 2015. Hydroxytyrosol-derived compounds: a basis for the creation of new pharmacological agents for cancer prevention and therapy. Journal of Medicinal Chemistry 58, 90899107. doi: 10.1021/acs.jmedchem.5b00669.
Bernini, R, Merendino, N, Romani, A and Velotti, F 2013. Naturally occurring hydroxytyrosol: synthesis and anticancer potential. Current Medicinal Chemistry 20, 655670. doi: 10.2174/092986713804999367.
Bernini, R, Mincione, E, Barontini, M and Crisante, F 2008. Convenient synthesis of hydroxytyrosol and its lipophilic derivatives from tyrosol or homovanillyl alcohol. Journal of Agricultural and Food Chemistry 56, 88978904. doi: 10.1021/jf801558z.
Castillo, C, Hernández, J, Valverde, I, Pereira, V, Sotillo, J, Alonso, ML and Benedito, JL 2006. Plasma malonaldehyde (MDA) and total antioxidant status (TAS) during lactation in dairy cows. Research in Veterinary Science 80, 133139. doi: 10.1016/j.rvsc.2005.06.003.
Chen, J, Xu, J, Li, J, Du, L, Chen, T, Liu, P, Peng, S, Wang, M and Song, H 2015. Epigallocatechin-3-gallate attenuates lipopolysaccharide-induced mastitis in rats via suppressing MAPK mediated inflammatory responses and oxidative stress. International Immunopharmacology 26, 147152. doi: 10.1016/j.intimp.2015.03.025.
Contreras, GA and Sordillo, LM 2011. Lipid mobilization and inflammatory responses during the transition period of dairy cows. Comparative Immunology, Microbiology and Infectious Diseases 34, 281289. doi: 10.1016/j.cimid.2011.01.004.
Dipasquale, D, Basiricò, L, Morera, P, Primi, R, Tröscher, A and Bernabucci, U 2018. Anti-inflammatory effects of conjugated linoleic acid isomers and essential fatty acids in bovine mammary epithelial cells. Animal 9, 17. doi: 10.1017/S1751731117003676.
Fu, Y, Gao, R, Cao, Y, Guo, M, Wei, Z, Zhou, E, Li, Y, Yao, M, Yang, Z and Zhang, N 2014. Curcumin attenuates inflammatory responses by suppressing TLR4-mediated NFκB signaling pathway in lipopolysaccharide-induced mastitis in mice. International Immunopharmacology 20, 5458. doi: 10.1016/j.intimp.2014.01.024.
Gambacorta, A, Tofani, D, Bernini, R and Migliorini, A 2007. High yielding preparation of a stable precursor of hydroxytyrosol by total synthesis and from the natural glucoside oleuropein. Journal of Agricultural and Food Chemistry 55, 33863391. doi: 10.1021/jf063353b.
Harijith, A, Ebenezer, DL and Natarajan, V 2014. Reactive oxygen species at the crossroads of inflammasome and inflammation. Frontiers in Physiology 5, 1-11. doi: 10.3389/fphys.2014.00352.
Hu, T, He, XW, Jiang, JG and Xu, XL 2014. Hydroxytyrosol and its potential therapeutic effects. Journal of Agricultural and Food Chemistry 55, 33863391. doi: 10.1021/jf405820v.
Imperatori, F, Barlozzari, G, Scardigli, A, Romani, A, Macrì, G, Polinori, N, Bernini, R and Santi, L 2018. Leishmanicidal activity of green tea leaves and pomegranate peel extracts on L. infantum. Natural Product Research 32, 17. doi: 10.1080/14786419.2018.1481841.
Karamese, M, Guvendi, B, Karamese, S A, Cinar, I, Can, S, Erol, HS, Aydin, H, Gelen, V and Karakus, E 2016. The protective effects of epigallocatechin gallate on lipopolysaccharide-induced hepatotoxicity: an in vitro study on Hep3B cells. Iranian Journal of Basic Medical Sciences 19, 483489.
Kim, HS, Quon, MJ and Kim, JA 2014. New insights into the mechanisms of polyphenols beyond antioxidant properties; lessons from the green tea polyphenol, epigallocatechin 3-gallate. Redox Biology 2, 187-195. doi: 10.1016/j.redox.2013.12.022.
Li, D, Zhang, N, Cao, Y, Zhang, W, Su, G, Sun, Y, Liu, Z, Li, F, Liang, D,Liu, B, Guo, M, Yunhe, F, Zang, X and Yang, Z 2013. Emodin ameliorates lipopolysccharide-induced mastitis in mice by inhibiting activation of NFκB and MAPKs signal pathways. European Journal of Pharmacology 705, 7985. doi: 10.1016/j.ejphar.2013.02.021.
Liu, Z, Sun, L, Zhu, L, Jia, X, Li, X, Jia, H, Wang, Y, Weber, P, Long, J, and Liu, J 2007. Hydroxytyrosol protects retinal pigment epithelial cells from acrolein-induced oxidative stress and mitochondrial dysfunction. Journal of Neurochemistry 103, 26902700. doi: 10.1111/j.1471-4159.2007.04954.x.
Mastrogiovanni, F, Bernini, R, Basiricò, L, Bernabucci, U, Campo, M, Romani, A, Santi, L, and Lacetera, N 2018. Antioxidant and anti-inflammatory effects of pomegranate peel extracts on bovine mammary epithelial cells BME-UV1. Natural Product Research 2, 15. doi: 10.1080/14786419.2018.1508149.
Mota, MAD, Landim, JSP, Targino, TSS, da Silva, SFR, da Silva, SL and Pereira, MRP 2015. Evaluation of the anti-inflammatory and analgesic effects of green tea (Camellia sinensis) in mice. Acta Cirurgica Brasileira 30, 242246. doi: 10.1590/S0102-865020150040000002.
Peng, Y, Moritz, M, Han, X, Giddings, TH, Lyon, A, Kollman, J, Winey, M, Yates, J, Agard, DA, Drubin, DG and Barnes, G 2015. Interaction of CK1δ with γTuSC ensures proper microtubule assembly and spindle positioning. Molecular Biology of the Cell 26, 25052518. doi: 10.1091%2Fmbc.E14-12-1627.
Ramos-Gomez, M, Kwak, MK, Dolan, PM, Itoh, K, Yamamoto, M, Talalay, P and Kensler, TW 2001. Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice. Proceedings of the National Academy of Sciences 98, 34103415. doi: 10.1073/pnas.051618798.
Sun, L, Luo, C, Long, J, Wei, D and Liu, J 2006. Acrolein is a mitochondrial toxin: Effects on respiratory function and enzyme activities in isolated rat liver mitochondria. Mitochondrion 6, 136142. doi: 10.1016/j.mito.2006.04.003.
Tomas-Barberan, FA and Andres-Lacueva, C 2012. Polyphenols and health: current state and progress. Journal of Agriculture and Food Chemistry 60, 87738775. doi: 10.1021/jf300671j.
Tuzcu, M, Sahin, N, Karatepe, M, Cikim, G, Kilinc, U and Sahin, K 2008. Epigallocatechin-3-gallate supplementation can improve antioxidant status in stressed quail. British Poultry Science 49, 643648. doi: 10.1080/00071660802298336.
Vilaplana-Pérez, C, Auñón, D, García-Flores, LA and Gil-Izquierdo, A 2014. Hydroxytyrosol and potential uses in cardiovascular diseases, cancer, and AIDS. Frontiers in Nutrition 1, 118. doi: 10.3389%2Ffnut.2014.00018.
Visioli, F, Galli, C, Bornet, F, Mattei, A, Patelli, R, Galli, G and Caruso, D 2000. Olive oil phenolics are dose-dependently absorbed in humans. FEBS Letters 468, 159160. doi: 10.1016/S0014-5793(00)01216-3.
Visioli, F, Poli, A and Galli, C 2002. Antioxidant and other biological activities of phenols from olives and olive oil. Medicinal Research Reviews 22, 6575. doi: 10.1002/med.1028.
Warleta, F, Sánchez Quesada, C, Campos, M, Allouche, Y, Beltrán, G and Gaforio, JJ 2011. Hydroxytyrosol protects against oxidative DNA damage in human breast cells. Nutrients 3, 839857. doi: 10.3390%2Fnu3100839.
Wein, S, Beyer, B, Gohlke, A, Blank, R, Metges, CC and Wolffram, S 2016. Systemic absorption of catechins after intraruminal or intraduodenal application of a green tea extract in cows. PLoS ONE 11, e0159428. doi: 10.1371/journal.pone.0159428.
Zavizion, B, van Duffelen, M, Schaeffer, W and Politis, I. 1996. Establishment and characterization of a bovine mammary myoepithelial cell line. In Vitro Cellular & Developmental Biology - Animal 32, 149158. doi: 10.1007/BF02723680.
Zhang, HY, Wang, JY and Yao, HP 2014. Epigallocatechin-3-gallate attenuates lipopolysaccharide-induced inflammation in human retinal endothelial cells. International Journal of Ophthalmology 7, 408412. doi: 10.3980%2Fj.issn.2222-3959.2014.03.04.
Zhong, Y, Chiou, YS, Pan, MH and Shahidi, F 2012. Anti-inflammatory activity of lipophilic epigallocatechin gallate (EGCG) derivatives in LPS-stimulated murine macrophages. Food Chemistry 134, 742748. doi: 10.1016/j.foodchem.2012.02.172.
Zhu, L, Liu, Z, Feng, Z, Hao, J, Shen, W, Li, X, Sun, L, Sharman, E, Wang, Y, Wertz, K, Weber, P, Shi, X and Liu, J 2010. Hydroxytyrosol protects against oxidative damage by simultaneous activation of mitochondrial biogenesis and phase II detoxifying enzyme systems in retinal pigment epithelial cells. Journal of Nutritional Biochemistry 21, 10891098. doi: 10.1016/j.jnutbio.2009.09.006.
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