Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells

Abstract

Objectives: The aim of this study is to examine the anti-aging effect of Hibiscus sabdariffa (HS) against oxidative stress induced by hydrogen peroxide (H2O2) on immortalized human ovarian epithelial cells (IHOECs). Materials and Methods: The obtained IHOECs were combined with 12.5 mg/mL HS extracted by adding 1mL of distilled water (dH2O) and different concentrations of H2O2 for oxidative effect. IHOECs cell viability was increased with HS extract and determined by MTT test assay. Real-time polymerase chain reaction was conducted to amplify and detect apoptosis-related BAX and BCL2, stem cell markers SOX2 and OCT4, and antioxidative nuclear factor erythroid 2-related factor 2 (NRF2) gene expressions. For the statistical analysis Kruskal-Wallis analysis and Mann-Whitney U test were used in SPSS program. Results: The increase in HS in cell proliferation was 14%. While there is no significant increase of cell viability in the combined group with HS and H2O2, there were only statistically significant groups are 200, 800, 900 and, 1,000 μl. It was indicated that HS has apoptotic, stem cell promotion and antioxidative effects on high H2O2 concentrations of these groups (p<0.05). Conclusion: In combined groups with both HS and H2O2 induced oxidative stress, HS increased the expression of proapoptotic gene BAX and decreased antiapoptotic gene BCL2. Although, the remaining cells which are abundantly stem cells expressed genes OCT4 and SOX2 remarkably decreased, at high H2O2 concentration opposite effects were observed. NRF2 gene expression also increased in this high oxidative stress.

Keywords

• Hibiscus sabdariffa
• oxidative stress
• apoptosis
• stem cells

Hibiscus sabdariffa’nın Ölümsüz İnsan Over Epitel Hücreleri Üzerindeki Yaşlanma Karşıtı Etkileri

Abstract

Amaç: Bu çalışmanın amacı, Hibiscus sabdariffa’nın (HS) hidrojen peroksitin (H2O2) ölümsüzleştirilmiş insan yumurtalık epitel hücreleri (IHOEC) üzerinde neden olduğu oksidatif strese karşı yaşlanma karşıtı etkisini incelemektir. Gereç ve Yöntem: Elde edilen IHOEC’ler, oksidatif etki için 1 mL distile su (dH2O) ve farklı konsantrasyonlarda H2O2 eklenerek ekstrakte edilen 12,5 mg/mL HS ile kombine olarak denendi. IHOEC’lerin hücre canlılığının, HS ekstraktı ile arttığı MTT testi ile belirlendi. Apoptozla ilişkili BAX ve BCL2’yi; kök hücre belirteçleri olan SOX2 ve OCT4’ü; antioksidatif belirteç olan nükleer faktör eritroid 2 ile ilişkili faktör 2 (NRF2) gen ifadelerini belirlemek için gerçek zamanlı polimeraz zincir reaksiyonu yöntemi kullanıldı. İstatistiksel analiz için SPSS programında Kruskal-Wallis analizi ve Mann-Whitney U testi kullanıldı. Bulgular: Hücre proliferasyonunda HS’deki artış %14 idi. HS ve H2O2 ile kombine edilen grupta hücre canlılığında anlamlı bir artış görülmezken, sadece 200, 800, 900 ve 1,000 μl’lik gruplar istatistiksel olarak anlamlıydı. HS’nin bu grupların yüksek H2O2 konsantrasyonları üzerinde apoptotik, kök hücre teşvik edici ve antioksidatif etkilere sahip olduğu belirtildi (p<0,05). Sonuç: Hem HS hem de H2O2 kaynaklı oksidatif stresin olduğu kombine gruplarda HS, proapoptotik gen BAX’ın ekspresyonunu artırdı ve antiapoptotik gen BCL2’yi azalttı. Geriye kalan kök hücre bakımından zengin kültürde OCT4 ve SOX2 genlerini ekspresyonu azalırken, yüksek H2O2 konsantrasyonunda tersine bir etki gözlendi. Bu yüksek oksidatif streste NRF2 gen ekspresyonu da artış gösterdi.

Keywords

• Hibiscus sabdariffa
• oksidatif stres
• apoptoz
• kök hücreler

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