Olive Leaf Extracts Protect Cardiomyocytes against 4-Hydroxynonenal-Induced Toxicity In Vitro: Comparison with Oleuropein, Hydroxytyrosol, and Quercetin

Elif Burcu Bali; Volkan Ergin; Lucia Rackova; Oğuz Bayraktar; Nurgün Küçükboyacı; Çimen Karasu

doi:10.1055/s-0034-1382881

Planta Medica, Inhaltsverzeichnis

Planta Med 2014; 80(12): 984-992
DOI: 10.1055/s-0034-1382881

Biological and Pharmacological Activity

Original Papers

Georg Thieme Verlag KG Stuttgart · New York

Olive Leaf Extracts Protect Cardiomyocytes against 4-Hydroxynonenal-Induced Toxicity In Vitro: Comparison with Oleuropein, Hydroxytyrosol, and Quercetin

Autor*innen

Elif Burcu Bali^*

¹Cellular Stress Response & Signal Transduction Research Laboratory, Department of Medical Pharmacology, Faculty of Medicine, Gazi University, and Farmasens Biotech Co., Gazi TechnoPark, Ankara, Turkey
Volkan Ergin^*

²Department of Medical Biology & Genetics, Faculty of Medicine, Gazi University, Ankara, Turkey
Lucia Rackova

³Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovak Republic
Oğuz Bayraktar

⁴Department of Chemical & Biochemical Engineering, Izmir Institute of Technology, and DUAG Co., İYTE, İzmir, Turkey
Nurgün Küçükboyacı

⁵Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Ankara, Turkey
Çimen Karasu

¹Cellular Stress Response & Signal Transduction Research Laboratory, Department of Medical Pharmacology, Faculty of Medicine, Gazi University, and Farmasens Biotech Co., Gazi TechnoPark, Ankara, Turkey

Abstract

Olive (Olea europaea) leaf, an important traditional herbal medicine, displays cardioprotection that may be related to the cellular redox modulating effects of its polyphenolic constituents. This study was undertaken to investigate the protective effect of the ethanolic and methanolic extracts of olive leaves compared to the effects of oleuropein, hydroxytyrosol, and quercetin as a positive standard in a carbonyl compound (4-hydroxynonenal)-induced model of oxidative damage to rat cardiomyocytes (H9c2). Cell viability was detected by the MTT assay; reactive oxygen species production was assessed by the 2′,7′-dichlorodihydrofluorescein diacetate method, and the mitochondrial membrane potential was determined using a JC-1 dye kit. Phospho-Hsp27 (Ser82), phospho-MAPKAPK-2 (Thr334), phospho-c-Jun (Ser73), cleaved-caspase-3 (cl-CASP3) (Asp175), and phospho-SAPK/JNK (Thr183/Tyr185) were measured by Western blotting. The ethanolic and methanolic extracts of olive leaves inhibited 4-hydroxynonenal-induced apoptosis, characterized by increased reactive oxygen species production, impaired viability (LD₅₀: 25 µM), mitochondrial dysfunction, and activation of pro-apoptotic cl-CASP3. The ethanolic and methanolic extracts of olive leaves also inhibited 4-hydroxynonenal-induced phosphorylation of stress-activated transcription factors, and the effects of extracts on p-SAPK/JNK, p-Hsp27, and p-MAPKAPK-2 were found to be concentration-dependent and comparable with oleuropein, hydroxytyrosol, and quercetin. While the methanolic extract downregulated 4-hydroxynonenal-induced p-MAPKAPK-2 and p-c-Jun more than the ethanolic extract, it exerted a less inhibitory effect than the ethanolic extract on 4-hydroxynonenal-induced p-SAPK/JNK and p-Hsp27. cl-CASP3 and p-Hsp27 were attenuated, especially by quercetin. Experiments showed a predominant reactive oxygen species inhibitory and mitochondrial protecting ability at a concentration of 1–10 µg/mL of each extract, oleuropein, hydroxytyrosol, and quercetin. The ethanolic extract of olive leaves, which contains larger amounts of oleuropein, hydroxytyrosol, verbascoside, luteolin, and quercetin (by HPLC) than the methanolic one, has more protecting ability on cardiomyocyte viability than the methanolic extract or each phenolic compound against 4-hydroxynonenal-induced carbonyl stress and toxicity.

Key words

Olea europaea L. - Oleaceae - olive leaf extract - cardiomyocytes - 4-hydroxynonenal - oleuropein - hydroxytyrosol - quercetin

Volltext

Referenzen

References
1 Poli G, Schaur RJ, Siems WG, Leonarduzzi G. 4-hydroxynonenal: a membrane lipid oxidation product of medicinal interest. Med Res Rev 2008; 28: 569-631
2 Uchida K, Stadtman ER. Modification of histidine residues in proteins by reaction with 4-hydroxynonenal. Proc Natl Acad Sci U S A 1992; 89: 4544-4548
3 Szapacs ME, Riggins JN, Zimmerman LJ, Liebler DC. Covalent adduction of human serum albumin by 4-hydroxy-2-nonenal: kinetic analysis of competing alkylation reactions. Biochemistry 2006; 45: 10521-10528
4 Grune T, Davies KJ. The proteasomal system and HNE-modified proteins. Mol Aspects Med 2003; 24: 195-204
5 Galvani S, Coatrieux C, Elbaz M, Grazide MH, Thiers JC, Parini A, Uchida K, Kamar N, Rostaing L, Baltas M, Salvayre R, Nègre-Salvayre A. Carbonyl scavenger and antiatherogenic effects of hydrazine derivatives. Free Radic Biol Med 2008; 45: 1457-1467
6 Miller DM, Singh IN, Wang JA, Hall ED. Administration of the Nrf2-ARE activators sulforaphane and carnosic acid attenuates 4-hydroxy-2-nonenal-induced mitochondrial dysfunction ex vivo . Free Radic Biol Med 2013; 57: 1-9
7 Karasu C. Glycoxidative stress and cardiovascular complications in experimentally-induced diabetes: effects of antioxidant treatment. Open Cardiovasc Med J 2010; 4: 240-256
8 Sakul A, Cumaoğlu A, Aydin E, Ari N, Dilsiz N, Karasu C. Age- and diabetes-induced regulation of oxidative protein modification in rat brain and peripheral tissues: consequences of treatment with antioxidant pyridoindole. Exp Gerontol 2013; 48: 476-484
9 Qin F, Simeone M, Patel R. Inhibition of NADPH oxidase reduces myocardial oxidative stress and apoptosis and improves cardiac function in heart failure after myocardial infarction. Free Radic Biol Med 2007; 43: 271-281
10 Anderson EJ, Katunga LA, Willis MS. Mitochondria as a source and target of lipid peroxidation products in healthy and diseased heart. Clin Exp Pharmacol Physiol 2012; 39: 179-193
11 He L, Liu B, Dai Z, Zhang HF, Zhang YS, Luo XJ, Ma QL, Peng J. Alpha lipoic acid protects heart against myocardial ischemia-reperfusion injury through a mechanism involving aldehyde dehydrogenase 2 activation. Eur J Pharmacol 2012; 678: 32-38
12 Bayçin D, Altiok E, Ulkü S, Bayraktar O. Adsorption of olive leaf (Olea europaea L.) antioxidants on silk fibroin. J Agric Food Chem 2007; 55: 1227-1236
13 Altıok E, Baycin D, Bayraktar O, Semra Ulkü S. Isolation of polyphenols from the extracts of olive leaves (Olea europaea L.) by adsorption on silk fibroin. Sep Purif Technol 2008; 62: 342-348
14 Dekanski D, Janicijevic-Hudomal S, Tadic V, Markovic G, Arsic I, Mitrovic DM. Phytochemical analysis and gastroprotective activity of an olive leaf extract. J Serb Chem Soc 2009; 74: 367-377
15 Mylonaki S, Kiassos E, Makris DP, Kefalas P. Optimisation of the extraction of olive (Olea europaea) leaf phenolics using water/ethanol-based solvent systems and response surface methodology. Anal Bioanal Chem 2008; 392: 977-985
16 Koca U, Süntar I, Akkol EK, Yilmazer D, Alper M. Wound repair potential of Olea europaea L. leaf extracts revealed by in vivo experimental models and comparative evaluation of the extractsʼ antioxidant activity. J Med Food 2011; 14: 140-146
17 Poudyal H, Campbell F, Brown L. Olive leaf extract attenuates cardiac, hepatic, and metabolic changes in high carbohydrate-, high fat-fed rats. J Nutr 2010; 140: 946-953
18 Manna C, Migliardi V, Golino P, Scognamiglio A, Galletti P, Chiariello M, Zappia V. Oleuropein prevents oxidative myocardial injury induced by ischemia and reperfusion. J Nutr Biochem 2004; 15: 461-466
19 Cumaoğlu A, Rackova L, Stefek M, Kartal M, Maechler P, Karasu C. Effects of olive leaf polyphenols against H₂O₂ toxicity in insulin secreting β-cells. Acta Biochim Pol 2011; 58: 45-50
20 Cumaoğlu A, Ari N, Kartal M, Karasu C. Polyphenolic extracts from Olea europaea L. protect against cytokine-induced β-cell damage through maintenance of redox homeostasis. Rejuvenation Res 2011; 14: 325-334
21 Stefek M, Karasu C. Eye lens in aging and diabetes: effect of quercetin. Rejuvenation Res 2011; 14: 525-534
22 Hur SJ, Lee SJ, Kim DH, Chun SC, Lee SK. Onion extract structural changes during in vitro digestion and its potential antioxidant effect on brain lipids obtained from low- and high-fat-fed mice. Free Radic Res 2013; 47: 1009-1015
23 Budas GR, Disatnik MH, Chen CH, Mochly-Rosen D. Activation of aldehyde dehydrogenase 2 (ALDH2) confers cardioprotection in protein kinase C epsilon (PKCvarepsilon) knockout mice. J Mol Cell Cardiol 2010; 48: 757-764
24 Ergin V, Hariry RE, Karasu C. Carbonyl stress in aging process: role of vitamins and phytochemicals as redox regulators. Aging Dis 2013; 4: 276-294
25 Liao J, Sun A, Xie Y, Isse T, Kawamoto T, Zou Y, Ge J. Aldehyde dehydrogenase-2 deficiency aggravates cardiac dysfunction elicited by endoplasmic reticulum stress induction. Mol Med 2012; 18: 785-793
26 Wei J, Wang W, Chopra I, Li HF, Dougherty CJ, Adi J, Adi N, Wang H, Webster KA. c-Jun N-terminal kinase (JNK-1) confers protection against brief but not extended ischemia during acute myocardial infarction. J Biol Chem 2011; 286: 13995-14006
27 Rouse J, Cohen P, Trigon S, Morange M, Alonso-Llamazares A, Zamanillo D, Hunt T, Nebreda AR. A novel kinase cascade triggered by stress and heat shock that stimulates MAPKAP kinase-2 and phosphorylation of the small heat shock proteins. Cell 1994; 78: 1027-1037
28 Mehlen P, Arrigo AP. The serum-induced phosphorylation of mammalian hsp27 correlates with changes in its intracellular localization and levels of oligomerization. Eur J Biochem 1994; 221: 327-334
29 Monastyrskaya EA, Andreeva LV, Duchen MR, Wiegant F, Bayda LA, Manukhina EB, Malyshev IY. Adaptation to heat of cardiomyoblasts in culture protects them against heat shock: role of nitric oxide and heat shock proteins. Biochemistry (Mosc) 2003; 68: 816-821
30 Kutuk O, Poli G, Basaga H. Resveratrol protects against 4-hydroxynonenal-induced apoptosis by blocking JNK and c-JUN/AP-1 signaling. Toxicol Sci 2006; 90: 120-132
31 Jang YJ, Kim J, Shim J, Kim J, Byun S, Oak MH, Lee KW, Lee HJ. Kaempferol attenuates 4-hydroxynonenal-induced apoptosis in PC12 cells by directly inhibiting NADPH oxidase. J Pharmacol Exp Ther 2011; 337: 747-754
32 Jin HB, Yang YB, Song YL, Zhang YC, Li YR. Protective roles of quercetin in acute myocardial ischemia and reperfusion injury in rats. Mol Biol Rep 2012; 39: 11005-11009
33 Yan L, Zhang JD, Wang B, Lv YJ, Jiang H, Liu GL, Qiao Y, Ren M, Guo XF. Quercetin inhibits left ventricular hypertrophy in spontaneously hypertensive rats and inhibits angiotensin II-induced H9c2 cells hypertrophy by enhancing PPAR-γ expression and suppressing AP-1 activity. PLoS One 2013; 8: e72548
34 Khreiss T, József L, Hossain S, Chan JS, Potempa LA, Filep JG. Loss of pentameric symmetry of C-reactive protein is associated with delayed apoptosis of human neutrophils. J Biol Chem 2002; 277: 40775-40781
35 Li F, Wu JH, Wang QH, Shu YL, Wan CW, Chan CO, Kam-Wah Mok D, Chan SW. Gui-ling-gao, a traditional Chinese functional food, prevents oxidative stress-induced apoptosis in H9c2 cardiomyocytes. Food Funct 2013; 4: 745-753
36 Angeloni C, Spencer JP, Leoncini E, Biagi PL, Hrelia S. Role of quercetin and its in vivo metabolites in protecting H9c2 cells against oxidative stress. Biochimie 2007; 89: 73-82
37 de Bock M, Derraik JG, Brennan CM, Biggs JB, Morgan PE, Hodgkinson SC, Hofman PL, Cutfield WS. Olive (Olea europaea L.) leaf polyphenols improve insulin sensitivity in middle-aged overweight men: a randomized, placebo-controlled, crossover trial. PLoS One 2013; 8: e57622
38 Janjic D, Wollheim CB. Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay. Diabetologia 1992; 35: 482-485
39 Royall JA, Ischiropoulos H. Evaluation of 2′,7′-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H₂O₂ in cultured endothelial cells. Arch Biochem Biophys 1993; 302: 348-355
40 Salvioli S, Ardizzoni A, Franceschi C, Cossarizza A. JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Lett 1997; 411: 77-82