PDF herunterladen
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

Elif Burcu Bali   *
1   Cellular 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   *
2   Department of Medical Biology & Genetics, Faculty of Medicine, Gazi University, Ankara, Turkey
,
Lucia Rackova
3   Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovak Republic
,
Oğuz Bayraktar
4   Department of Chemical & Biochemical Engineering, Izmir Institute of Technology, and DUAG Co., İYTE, İzmir, Turkey
,
Nurgün Küçükboyacı
5   Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Ankara, Turkey
,
Çimen Karasu
1   Cellular Stress Response & Signal Transduction Research Laboratory, Department of Medical Pharmacology, Faculty of Medicine, Gazi University, and Farmasens Biotech Co., Gazi TechnoPark, Ankara, Turkey
› Institutsangaben
Weitere Informationen

Publikationsverlauf

received 19. Januar 2014
revised 14. Mai 2014

accepted 16. Juni 2014

Publikationsdatum:
06. August 2014 (online)

Auch verfügbar auf
    Lizenzen und Reprints

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

* Both of these authors contributed equally to this study.


 

  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 4 Grune T, Davies KJ. The proteasomal system and HNE-modified proteins. Mol Aspects Med 2003; 24: 195-204
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 7 Karasu C. Glycoxidative stress and cardiovascular complications in experimentally-induced diabetes: effects of antioxidant treatment. Open Cardiovasc Med J 2010; 4: 240-256
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    PubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 21 Stefek M, Karasu C. Eye lens in aging and diabetes: effect of quercetin. Rejuvenation Res 2011; 14: 525-534
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    PubMedSuche in Google Scholar
  • 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
    PubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    PubMedSuche in Google Scholar
  • 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
    PubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 38 Janjic D, Wollheim CB. Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay. Diabetologia 1992; 35: 482-485
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar
  • 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
    CrossrefPubMedSuche in Google Scholar