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

Yıl 2020, Cilt: 4 Sayı: 1, 21 - 28, 02.03.2020
https://doi.org/10.30565/medalanya.607989

Öz

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. 

Destekleyen Kurum

Hatay Mustafa Kemal Üniversitesi, Bilimsel Araştırma Projeleri Koordinatörlüğü

Proje Numarası

18.YL.087

Teşekkür

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.

Kaynakça

  • 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

Yıl 2020, Cilt: 4 Sayı: 1, 21 - 28, 02.03.2020
https://doi.org/10.30565/medalanya.607989

Öz

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.

Proje Numarası

18.YL.087

Kaynakça

  • 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
Toplam 30 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Klinik Tıp Bilimleri
Bölüm Araştırma Makalesi
Yazarlar

Hayriye Zehra Ulutaş Bu kişi benim 0000-0001-6553-3420

Gülay Gülbol Duran 0000-0002-4672-2960

Proje Numarası 18.YL.087
Yayımlanma Tarihi 2 Mart 2020
Gönderilme Tarihi 21 Ağustos 2019
Kabul Tarihi 28 Kasım 2019
Yayımlandığı Sayı Yıl 2020 Cilt: 4 Sayı: 1

Kaynak Göster

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.

9705 

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