Antioxidant Effects of Myrtus communis L.’s Essential Oils in BEAS-2B Cells Induced by Oxidative Stress with Hydrogen Peroxide

Hayriye Zehra Ulutaş; Gülay Gülbol Duran

doi:10.30565/medalanya.607989

Research Article

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Hidrojen Peroksit ile Okdisatif Stresin İndüklendiği BEAS-2B Hücrelerinde Myrtus communis L. Esansiyel Yağının Antioksidan Etkilerinin Araştırılması

Year 2020, Volume: 4 Issue: 1, 21 - 28, 02.03.2020

Hayriye Zehra Ulutaş Gülay Gülbol Duran

https://doi.org/10.30565/medalanya.607989

Cited By: 1

Abstract

Amaç: Bu çalışmada, Myrtus communis L. esansiyel yağının, in vitro olarak hidrojen peroksit ile oksidatif strese uğratılmış insan bronşiyal epitel hücre hattında (BEAS-2B) antioksidan etkilerinin olup olmadığı ve apoptotik yolaklar üzerindeki etkileri araştırılmıştır.
Materyal ve Metod: BEAS-2B hücrelerinde H2O2 ile M. communis L. esansiyel yağının uygun dozlarının belirlenmesinde MTT yöntemi kullanılmıştır. Oksidatif stres oluşumu MDA düzeyi ölçülerek belirlenmiştir. Hücreler; oksidatif strese maruz bırakılan grup (H2O2), tedavili grup (H2O2+M. communis L) ve kontrol grubu olmak üzere üç gruba ayrılmıştır. Her üç grupta MDA düzeyleri ölçülmüş ve apoptotik etkilerin saptanması amacıyla Kaspaz 3, Kaspaz 8, Kaspaz 9 ile p21 genlerinin ekspresyon düzeyleri RT-PCR yöntemi ile saptanmıştır.
Bulgular: MTT testi ile uygun dozlar H2O2 için 40 μM, M. communis L. yağı için 15.625 μg/ml olarak saptanmıştır. Sağlıklı hücre grubuyla karşılaştırıldığında, 40 μM H2O2 maruziyeti uygulanan grupta MDA düzeyinde anlamlı artış gözlenirken (p=0,0005), M. communis L. esansiyel yağı ile tedavi edilen gruptaki MDA düzeyi kontrol grubu ile benzer düzeyde (p>0.05) bulunmuştur. 40 μM H2O2 uygulanan hücrelerde Kaspaz 3, Kaspaz 8 ve p21 genlerinin ekspresyon seviyelerinin sağlıklı hücre grubuna göre anlamlı bir şekilde arttığı (sırasıyla p=0.001, p=0.017 ve p=0,0003), Kaspaz 9 gen ekspresyon seviyesinin değişmediği (p=0.8) saptanmıştır. M. communis L. yağı ile tedavi edilen hücrelerde, oksidatif stres modelinin oluşturulduğu gruba göre Kaspaz 3 gen ekspresyon seviyesinin anlamlı şekilde azaldığı saptanmıştır (p=0.00007).
Sonuç: Çalışmamızda, M. communis L. esansiyel yağının MDA seviyesini güçlü şekilde azaltıcı etkisi olduğu, ayrıca apopitozu durdurucu bir etki göstererek, teröpatik ajan olarak potansiyeli olduğu gösterildi. Bu bağlamda M. communis L. yağının güçlü bir antioksidan etkiye sahip olduğu, oksidatif stresten kaynaklanan apoptozisin durdurulmasında etkili olduğu düşünülmektedir.

Keywords

Myrtus communis L, Apopitoz, BEAS-2B, Oksidatif Stres

Supporting Institution

Hatay Mustafa Kemal Üniversitesi, Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

18.YL.087

Thanks

Myrtus communis L esansiyel yağının temin edilmesindeki katkılarından dolayı Hatay Mustafa Kemal Üniversitesi, Ziraat Fakültesi, Tıbbi Bitkiler Anabilim Dalı öğretim üyesi Doç. Dr. Durmuş Alparslan KAYA’ya teşekkür ederiz.

References

1. Hayder N, Abdelwahed A, Kilani S, Ammar RB, Mahmoud A, Ghedira K, et al. Anti-genotoxic and free-radical scavenging activities of extracts from (Tunisian) Myrtus communis. Mutat Res Genet Toxicol Environ Mutagen. 2004;564(1):89-95. PMID:15474415
2. Aydın C, Özcan MM. Determination of nutritional and physical properties of myrtle (Myrtus communis L.) fruits growing wild in Turkey. J Food Eng. 2007;79(2):453-8. DOI: 10.1016/j.jfoodeng.2006.02.008
3. Mansouri S, Foroumadi A, Ghaneie T, Najar AG. Antibacterial Activity of the Crude Extracts and Fractionated Constituents of Myrtus communis. Pharmaceutical Bio. 2001;39(5):399-401. DOI: 10.1076/phbi.39.5.399.5889
4. Cakir A. Essential oil and fatty acid composition of the fruits of Hippophae rhamnoides L. (Sea Buckthorn) and Myrtus communis L. from Turkey. Biochem Syst Ecol. 2004;32(9):809-16. DOI: 10.1016/j.bse.2003.11.010
5. Gardeli C, Vassiliki P, Athanasios M, Kibouris T, Komaitis M. Essential oil composition of Pistacia lentiscus L. and Myrtus communis L.: Evaluation of antioxidant capacity of methanolic extracts. Food Chem. 2008;107(3):1120-30. DOI: 10.1016/j.foodchem.2007.09.036
6. Smith MA, Perry G, Pryor WA. Causes and consequences of oxidative stress in Alzheimer’s disease 1,2 1Guest Editors: Mark A. Smith and George Perry 2This article is part of a series of reviews on “Causes and Consequences of Oxidative Stress in Alzheimer’s Disease.” Free Radic Biol Med. 2002;32(11):1049. PMID:12031888
7. Olinski R, Gackowski D, Foksinski M, Rozalski R, Roszkowski K, Jaruga P. Oxidative DNA damage: assessment of the role in carcinogenesis, atherosclerosis, and acquired immunodeficiency syndrome1. Free Radic Biol Med. 2002;33(2):192-200. DOI: 10.1016/S0891-5849(02)00878-X
8. Mimica-Dukić N, Božin B, Soković M, Mihajlovic B, Matavulj M. Antimicrobial and Antioxidant Activities of Three Mentha Species Essential Oils. Planta Med. 2003;69(5):413-9. DOI: 10.1055/s-2003-39704
9. Hayder N, Skandrani I, Kilani S, Bouhlel I, Abdelwahed A, Ben Ammar R, et al. Antimutagenic activity of Myrtus communis L. using the Salmonella microsome assay. S Afr J Bot.2008;74(1):121-5. DOI: 10.1016/j.sajb.2007.10.001
10. Sushil JK. Membrane lipid peroxidation in erythrocytes of the newborn. Clin Chim Acta. 1986;161(3):301-6. DOI: 10.1016/0009-8981(86)90014-8
11. Masoudi M, Miraj S, Rafieian-Kopaei M. Comparison of the Effects of Myrtus Communis L, Berberis Vulgaris and Metronidazole Vaginal Gel alone for the Treatment of Bacterial Vaginosis. J Clin Diagn Res. 2016;10(3):QC04-QC7. PMID: 27134945
12. Kumar R, Phaneendra P, Bodhanapu S, Rahiman F, Mohamed NK, Tamizmani T. Antioxidant and hepatoprotective activity of the aqueous extract of Myrtus communis (myrtle) leaves. Pharmacololgy. 2011;1083-90.
13. Rossi A, Di Paola R, Mazzon E, Genovese T, Caminiti R, Bramanti P, et al. Myrtucommulone from <em>Myrtus communis</em> Exhibits Potent Anti-Inflammatory Effectiveness in Vivo. J Pharmacol Exp Ther.2009;329(1):76-86. DOI: 10.1124/jpet.108.143214
14. Mimika-Dukić N, Bugarin D, Grbović S, Mitić-Ćulafić D, Vuković-Gačić B, Orčić D, et al. Essential Oil of Myrtus communis L. as a Potential Antioxidant and Antimutagenic Agents. Molecules. 2010;15(4):2759-70. DOI: 10.3390/molecules15042759
15. de Lavor ÉM, Fernandes AWC, de Andrade Teles RB, Leal AEBP, de Oliveira Júnior RG, Gama E Silva M, et al. Essential Oils and Their Major Compounds in the Treatment of Chronic Inflammation: A Review of Antioxidant Potential in Preclinical Studies and Molecular Mechanisms. Oxid Med Cell Longev. 2018;2018:6468593-. PMID:30671173
16. Lu JM, Lin PH, Yao Q, Chen C. Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems. J Cell Mol Med. 2010;14(4):840-60. PMID:19754673
17. Dennis JM, Witting PK. Protective Role for Antioxidants in Acute Kidney Disease. Nutrients. 2017;9(7):718. DOI: 10.3390/nu9070718
18. Park JE, Yang JH, Yoon SJ, Lee JH, Yang ES, Park JW. Lipid peroxidation-mediated cytotoxicity and DNA damage in U937 cells. Biochimie. 2002;84(12):1199-205. DOI: 10.1016/S0300-9084(02)00039-1
19. Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncol. 2012;2012:137289. DOI: 10.5402/2012/137289
20. Saha SK, Lee SB, Won J, Choi HY, Kim K, Yang G-M, et al. Correlation between Oxidative Stress, Nutrition, and Cancer Initiation. Int J Mol Sci. 2017;18(7):1544. DOI: 10.3390/ijms18071544
21. Romani A, Coinu R, Carta S, Pinelli P, Galardi C, Vincieri FF, et al. Evaluation of antioxidant effect of different extracts of Myrtus communis L. Free Radic Res. 2004;38(1):97-103. DOI: 10.1080/10715760310001625609
22. Ines S, Ines B, Wissem B, Mohamed BS, Nawel H, Dijoux-Franca MG, et al. In vitro antioxidant and antigenotoxic potentials of 3,5-O-di-galloylquinic acid extracted from Myrtus communis leaves and modulation of cell gene expression by H2O2. J Appl Toxicol. 2012;32(5):333-41. DOI: 10.1002/jat.1655
23. Turhan N, Celik H, Duvan Cİ, Onaran Y, Aydın M, Armutcu F. Investigation of oxidative balance in patients with dysmenorrhea by multiple serum markers. J Turk Ger Gynecol Assoc. 2012;13(4):233-6. PMID: 24592048
24. Fadaei B, Ahmadvand H, Ghasemi M, Mirzaie S. Inhibitory effects of myrtle (Myrtus communis L.) leaves hydroalcoholic extract on LDL oxidation in vitro. J Chem Pharm Res. 2015;42-6.
25. Bajpai VK, Agrawal P, Bang BH, Park Y-H. Phytochemical analysis, antioxidant and antilipid peroxidation effects of a medicinal plant, Adhatoda vasica. Front Life Sci. 2015;8(3):305-12. DOI: 10.1080/21553769.2014.1002943
26. Sahreen S, Khan MR, Khan RA. Comprehensive assessment of phenolics and antiradical potential of Rumex hastatus D. Don. roots. BMC Complement and Altern Med. 2014;14(1). DOI: 10.1186/1472-6882-14-47
27. Miguel MG. Antioxidant activity of medicinal and aromatic plants. A review. Flavour Fragr J. 2010;25(5):291-312. DOI: 10.1002/ffj.1961
28. Gonçalves S, Gomes D, Costa P, Romano A. The phenolic content and antioxidant activity of infusions from Mediterranean medicinal plants. Ind Crops Prod. 2013;43:465-71. DOI: 10.1016/j.indcrop.2012.07.066
29. Fernald K, Kurokawa M. Evading apoptosis in cancer. Trends Cell Biol.2013;23(12):620-33. DOI: 10.1016/j.tcb.2013.07.006
30. Tretiakova I, Blaesius D, Maxia L, Wesselborg S, Schulze-Osthoff K, Cinatl J, et al. Myrtucommulone from Myrtus communis induces apoptosis in cancer cells via the mitochondrial pathway involving caspase-9. Apoptosis. 2007;13(1):119-31. DOI: 10.1007/s10495-007-0150-0

Antioxidant Effects of Myrtus communis L.’s Essential Oils in BEAS-2B Cells Induced by Oxidative Stress with Hydrogen Peroxide

Year 2020, Volume: 4 Issue: 1, 21 - 28, 02.03.2020

Hayriye Zehra Ulutaş Gülay Gülbol Duran

https://doi.org/10.30565/medalanya.607989

Cited By: 1

Abstract

Aim: In this study, the effects of Myrtus communis L. essential oil on the human bronchial epithelial cell line (BEAS-2B) exposed to oxidative stress with hydrogen peroxide were investigated and their effects on apoptotic pathways.

Materials and Methods: The MTT 3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide method was used to determine the appropriate doses of hydrogen peroxide (H2O2) and M. communis L.’s essential oil in BEAS-2B cells. Oxidative stress formation was determined by measuring malondialdehyde (MDA) level. The cells were divided into three groups: the group exposed to oxidative stress (group with H2O2), the treatment group (H2O2 + M. communis L.’s essential oil) and the control group. MDA levels were measured in all three groups and expression levels of Caspase 3, Caspase 8, Caspase 9 and p21 genes were determined by RT-PCR method in order to detect apoptotic effects.

Results: According to MTT test results, the appropriate doses were 40 μM for H2O2 and 15.625 μg/ml for M. communis L.’s essential oil. MDA levels were significantly increased in the group treated with 40 μM H2O2 when compared with the healthy cell group (p=0.0005). In the group treated with essential oil of M. communis L., MDA level was found similar to the control group (p>0.05). Expression levels of Caspase 3, Caspase 8 and p21 genes were significantly increased in cells where H2O2 was adminisrated at 40 μM concentrations compared to healthy cell group (p=0.001, p=0.017 and p=0.0003, respectively). However, Caspase 9 gene expression level did not change significantly (p=0.8). Compared to the group in which the oxidative stress model was established, it was found that Caspase 3 gene expression level decreased significantly in the cells treated with M. communis L.’s oil (p=0.00007).

Conclusion: In our study, it was shown that the essential oil of M. communis L. strongly decreased MDA levels and also had the potential to be a therapeutic agent due to its apoptotic inhibiting effect. M. communis L. has a strong antioxidant effect and is thought to be effective in stopping apoptosis caused by oxidative stress.

Keywords

Myrtus communis L, Apoptosis, BEAS-2B, Oxidative Stress

Project Number

18.YL.087

References

1. Hayder N, Abdelwahed A, Kilani S, Ammar RB, Mahmoud A, Ghedira K, et al. Anti-genotoxic and free-radical scavenging activities of extracts from (Tunisian) Myrtus communis. Mutat Res Genet Toxicol Environ Mutagen. 2004;564(1):89-95. PMID:15474415
2. Aydın C, Özcan MM. Determination of nutritional and physical properties of myrtle (Myrtus communis L.) fruits growing wild in Turkey. J Food Eng. 2007;79(2):453-8. DOI: 10.1016/j.jfoodeng.2006.02.008
3. Mansouri S, Foroumadi A, Ghaneie T, Najar AG. Antibacterial Activity of the Crude Extracts and Fractionated Constituents of Myrtus communis. Pharmaceutical Bio. 2001;39(5):399-401. DOI: 10.1076/phbi.39.5.399.5889
4. Cakir A. Essential oil and fatty acid composition of the fruits of Hippophae rhamnoides L. (Sea Buckthorn) and Myrtus communis L. from Turkey. Biochem Syst Ecol. 2004;32(9):809-16. DOI: 10.1016/j.bse.2003.11.010
5. Gardeli C, Vassiliki P, Athanasios M, Kibouris T, Komaitis M. Essential oil composition of Pistacia lentiscus L. and Myrtus communis L.: Evaluation of antioxidant capacity of methanolic extracts. Food Chem. 2008;107(3):1120-30. DOI: 10.1016/j.foodchem.2007.09.036
6. Smith MA, Perry G, Pryor WA. Causes and consequences of oxidative stress in Alzheimer’s disease 1,2 1Guest Editors: Mark A. Smith and George Perry 2This article is part of a series of reviews on “Causes and Consequences of Oxidative Stress in Alzheimer’s Disease.” Free Radic Biol Med. 2002;32(11):1049. PMID:12031888
7. Olinski R, Gackowski D, Foksinski M, Rozalski R, Roszkowski K, Jaruga P. Oxidative DNA damage: assessment of the role in carcinogenesis, atherosclerosis, and acquired immunodeficiency syndrome1. Free Radic Biol Med. 2002;33(2):192-200. DOI: 10.1016/S0891-5849(02)00878-X
8. Mimica-Dukić N, Božin B, Soković M, Mihajlovic B, Matavulj M. Antimicrobial and Antioxidant Activities of Three Mentha Species Essential Oils. Planta Med. 2003;69(5):413-9. DOI: 10.1055/s-2003-39704
9. Hayder N, Skandrani I, Kilani S, Bouhlel I, Abdelwahed A, Ben Ammar R, et al. Antimutagenic activity of Myrtus communis L. using the Salmonella microsome assay. S Afr J Bot.2008;74(1):121-5. DOI: 10.1016/j.sajb.2007.10.001
10. Sushil JK. Membrane lipid peroxidation in erythrocytes of the newborn. Clin Chim Acta. 1986;161(3):301-6. DOI: 10.1016/0009-8981(86)90014-8
11. Masoudi M, Miraj S, Rafieian-Kopaei M. Comparison of the Effects of Myrtus Communis L, Berberis Vulgaris and Metronidazole Vaginal Gel alone for the Treatment of Bacterial Vaginosis. J Clin Diagn Res. 2016;10(3):QC04-QC7. PMID: 27134945
12. Kumar R, Phaneendra P, Bodhanapu S, Rahiman F, Mohamed NK, Tamizmani T. Antioxidant and hepatoprotective activity of the aqueous extract of Myrtus communis (myrtle) leaves. Pharmacololgy. 2011;1083-90.
13. Rossi A, Di Paola R, Mazzon E, Genovese T, Caminiti R, Bramanti P, et al. Myrtucommulone from <em>Myrtus communis</em> Exhibits Potent Anti-Inflammatory Effectiveness in Vivo. J Pharmacol Exp Ther.2009;329(1):76-86. DOI: 10.1124/jpet.108.143214
14. Mimika-Dukić N, Bugarin D, Grbović S, Mitić-Ćulafić D, Vuković-Gačić B, Orčić D, et al. Essential Oil of Myrtus communis L. as a Potential Antioxidant and Antimutagenic Agents. Molecules. 2010;15(4):2759-70. DOI: 10.3390/molecules15042759
15. de Lavor ÉM, Fernandes AWC, de Andrade Teles RB, Leal AEBP, de Oliveira Júnior RG, Gama E Silva M, et al. Essential Oils and Their Major Compounds in the Treatment of Chronic Inflammation: A Review of Antioxidant Potential in Preclinical Studies and Molecular Mechanisms. Oxid Med Cell Longev. 2018;2018:6468593-. PMID:30671173
16. Lu JM, Lin PH, Yao Q, Chen C. Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems. J Cell Mol Med. 2010;14(4):840-60. PMID:19754673
17. Dennis JM, Witting PK. Protective Role for Antioxidants in Acute Kidney Disease. Nutrients. 2017;9(7):718. DOI: 10.3390/nu9070718
18. Park JE, Yang JH, Yoon SJ, Lee JH, Yang ES, Park JW. Lipid peroxidation-mediated cytotoxicity and DNA damage in U937 cells. Biochimie. 2002;84(12):1199-205. DOI: 10.1016/S0300-9084(02)00039-1
19. Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncol. 2012;2012:137289. DOI: 10.5402/2012/137289
20. Saha SK, Lee SB, Won J, Choi HY, Kim K, Yang G-M, et al. Correlation between Oxidative Stress, Nutrition, and Cancer Initiation. Int J Mol Sci. 2017;18(7):1544. DOI: 10.3390/ijms18071544
21. Romani A, Coinu R, Carta S, Pinelli P, Galardi C, Vincieri FF, et al. Evaluation of antioxidant effect of different extracts of Myrtus communis L. Free Radic Res. 2004;38(1):97-103. DOI: 10.1080/10715760310001625609
22. Ines S, Ines B, Wissem B, Mohamed BS, Nawel H, Dijoux-Franca MG, et al. In vitro antioxidant and antigenotoxic potentials of 3,5-O-di-galloylquinic acid extracted from Myrtus communis leaves and modulation of cell gene expression by H2O2. J Appl Toxicol. 2012;32(5):333-41. DOI: 10.1002/jat.1655
23. Turhan N, Celik H, Duvan Cİ, Onaran Y, Aydın M, Armutcu F. Investigation of oxidative balance in patients with dysmenorrhea by multiple serum markers. J Turk Ger Gynecol Assoc. 2012;13(4):233-6. PMID: 24592048
24. Fadaei B, Ahmadvand H, Ghasemi M, Mirzaie S. Inhibitory effects of myrtle (Myrtus communis L.) leaves hydroalcoholic extract on LDL oxidation in vitro. J Chem Pharm Res. 2015;42-6.
25. Bajpai VK, Agrawal P, Bang BH, Park Y-H. Phytochemical analysis, antioxidant and antilipid peroxidation effects of a medicinal plant, Adhatoda vasica. Front Life Sci. 2015;8(3):305-12. DOI: 10.1080/21553769.2014.1002943
26. Sahreen S, Khan MR, Khan RA. Comprehensive assessment of phenolics and antiradical potential of Rumex hastatus D. Don. roots. BMC Complement and Altern Med. 2014;14(1). DOI: 10.1186/1472-6882-14-47
27. Miguel MG. Antioxidant activity of medicinal and aromatic plants. A review. Flavour Fragr J. 2010;25(5):291-312. DOI: 10.1002/ffj.1961
28. Gonçalves S, Gomes D, Costa P, Romano A. The phenolic content and antioxidant activity of infusions from Mediterranean medicinal plants. Ind Crops Prod. 2013;43:465-71. DOI: 10.1016/j.indcrop.2012.07.066
29. Fernald K, Kurokawa M. Evading apoptosis in cancer. Trends Cell Biol.2013;23(12):620-33. DOI: 10.1016/j.tcb.2013.07.006
30. Tretiakova I, Blaesius D, Maxia L, Wesselborg S, Schulze-Osthoff K, Cinatl J, et al. Myrtucommulone from Myrtus communis induces apoptosis in cancer cells via the mitochondrial pathway involving caspase-9. Apoptosis. 2007;13(1):119-31. DOI: 10.1007/s10495-007-0150-0

There are 30 citations in total.

Details

Primary Language	English
Subjects	Clinical Sciences
Journal Section	Research Article
Authors	Hayriye Zehra Ulutaş This is me 0000-0001-6553-3420 Gülay Gülbol Duran 0000-0002-4672-2960
Project Number	18.YL.087
Publication Date	March 2, 2020
Submission Date	August 21, 2019
Acceptance Date	November 28, 2019
Published in Issue	Year 2020 Volume: 4 Issue: 1

Cite

Vancouver	Ulutaş HZ, Gülbol Duran G. Antioxidant Effects of Myrtus communis L.’s Essential Oils in BEAS-2B Cells Induced by Oxidative Stress with Hydrogen Peroxide. Acta Med. Alanya. 2020;4(1):21-8.

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