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

Year 2020, , 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.

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

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

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