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

Year 2024, Volume: 77 Issue: 4, 374 - 380, 31.12.2024

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.

Ethical Statement

The cells used in the study are commercially available so ethics committee approval is not required. The study protocol was appropriate in accordance with the Declaration of Helsinki.

Supporting Institution

-

Project Number

-

Thanks

-

References

  • 1. Noll C, Raaf L, Planque C, et al. Protection and reversal of hepatic fibrosis by red wine polyphenols in hyperhomocysteinemic mice. J Nutr Biochem. 2011;22:856-864.
  • 2. Zhao W, Ji X, Zhang F, et al. Embryonic stem cell markers. Molecules. 2012;17:6196-6236.
  • 3. Sporn MB, Liby KT. NRF2 and cancer: the good, the bad and the importance of context. Nat Rev Cancer. 2012;12:564-571.
  • 4. da Silva PFL, Schumacher B. Principles of the Molecular and Cellular Mechanisms of Aging. J Invest Dermatol. 2021;141:951-960.
  • 5. Wang D, Nagata M, Matsumoto M, et al. Potential of Hibiscus sabdariffa L. and Hibiscus Acid to Reverse Skin Aging. Molecules. 2022;27:6076.
  • 6. Alkadi H. A Review on Free Radicals and Antioxidants. Infect Disord Drug Targets. 2020;20:16-26.
  • 7. Riaz G, Chopra R. A review on phytochemistry and therapeutic uses of Hibiscus sabdariffa L. Biomed Pharmacother. 2018;102:575-586.
  • 8. Ighodaro O. M., Akinloye O. A. First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine. 2018;54:287-293.
  • 9. Schuyer M, van der Burg ME, Henzen-Logmans SC, et al. Reduced expression of BAX is associated with poor prognosis in patients with epithelial ovarian cancer: a multifactorial analysis of TP53, p21, BAX and BCL-2. Br J Cancer. 2001;85:1359-67.
  • 10. Arnold K, Sarkar A, Yram MA, et al. Sox2(+) adult stem and progenitor cells are important for tissue regeneration and survival of mice. Cell Stem Cell. 2011;9:317-29.
  • 11. Pan GJ, Chang ZY, Schöler HR, et al. Stem cell pluripotency and transcription factor Oct4. Cell Res. 2002;12:321-329.
  • 12. Rojo de la Vega M, Chapman E, Zhang DD. NRF2 and the Hallmarks of Cancer. Cancer Cell. 2018;34:21-43.
  • 13. Menegon S, Columbano A, Giordano S. The Dual Roles of NRF2 in Cancer. Trends Mol Med. 2016;22:578-593.
  • 14. D’Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int. 2019;43:582-592.
  • 15. He F, Ru X, Wen T. NRF2, a Transcription Factor for Stress Response and Beyond. Int J Mol Sci. 2020;21:4777.
  • 16. Al-Hashimi, A.G. Antioxidant and Antibacterial Activities of Hibiscus sabdariffa L. Extracts. African Journal of Food Science. 2012;6:506-511.
  • 17. Li J, Lu YR, Lin IF, et al. Reversing UVB-induced photoaging with Hibiscus sabdariffa calyx aqueous extract. J Sci Food Agric. 2020;100:672-681.
  • 18. Malacrida A, Erriquez J, Hashemi M, et al. Evaluation of antitumoral effect of Hibiscus sabdariffa extract on human breast cancer cells. Biochem Biophys Rep. 2022;32:101353.
  • 19. Abdul Hamid Z, Lin Lin WH, Abdalla BJ, et al. The role of Hibiscus sabdariffa L. (Roselle) in maintenance of ex vivo murine bone marrow-derived hematopoietic stem cells. ScientificWorldJournal. 2014;2014:258192.
  • 20. Kaspar JW, Niture SK, Jaiswal AK. Nrf2:INrf2 (Keap1) signaling in oxidative stress. Free Radic Biol Med. 2009;47:1304-1309.
  • 21. Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem. 2009;284:13291-13295.
  • 22. Prasomthong J, Limpeanchob N, Daodee S, et al. Hibiscus sabdariffa extract improves hepatic steatosis, partially through IRS-1/Akt and Nrf2 signaling pathways in rats fed a high fat diet. Sci Rep. 2022;12:7022.
  • 23. Niture SK, Jaiswal AK. Nrf2 protein up-regulates antiapoptotic protein Bcl- 2 and prevents cellular apoptosis. J Biol Chem. 2012;287:9873-9886.
  • 24. Cengiz M, Ayhanci A, Akkemik E, et al. The role of Bax/Bcl-2 and Nrf2- Keap-1 signaling pathways in mediating the protective effect of boric acid on acrylamide-induced acute liver injury in rats. Life Sci. 2022;307:120864.
  • 25. Jaramillo MC, Zhang DD. The emerging role of the Nrf2-Keap1 signaling pathway in cancer. Genes Dev. 2013;27:2179-2191.
  • 26. Wu S, Lu H, Bai Y. Nrf2 in cancers: A double-edged sword. Cancer Med. 2019;8:2252-2267.

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

Year 2024, Volume: 77 Issue: 4, 374 - 380, 31.12.2024

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.

Ethical Statement

The cells used in the study are commercially available so ethics committee approval is not required. The study protocol was appropriate in accordance with the Declaration of Helsinki.

Supporting Institution

-

Project Number

-

Thanks

-

References

  • 1. Noll C, Raaf L, Planque C, et al. Protection and reversal of hepatic fibrosis by red wine polyphenols in hyperhomocysteinemic mice. J Nutr Biochem. 2011;22:856-864.
  • 2. Zhao W, Ji X, Zhang F, et al. Embryonic stem cell markers. Molecules. 2012;17:6196-6236.
  • 3. Sporn MB, Liby KT. NRF2 and cancer: the good, the bad and the importance of context. Nat Rev Cancer. 2012;12:564-571.
  • 4. da Silva PFL, Schumacher B. Principles of the Molecular and Cellular Mechanisms of Aging. J Invest Dermatol. 2021;141:951-960.
  • 5. Wang D, Nagata M, Matsumoto M, et al. Potential of Hibiscus sabdariffa L. and Hibiscus Acid to Reverse Skin Aging. Molecules. 2022;27:6076.
  • 6. Alkadi H. A Review on Free Radicals and Antioxidants. Infect Disord Drug Targets. 2020;20:16-26.
  • 7. Riaz G, Chopra R. A review on phytochemistry and therapeutic uses of Hibiscus sabdariffa L. Biomed Pharmacother. 2018;102:575-586.
  • 8. Ighodaro O. M., Akinloye O. A. First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine. 2018;54:287-293.
  • 9. Schuyer M, van der Burg ME, Henzen-Logmans SC, et al. Reduced expression of BAX is associated with poor prognosis in patients with epithelial ovarian cancer: a multifactorial analysis of TP53, p21, BAX and BCL-2. Br J Cancer. 2001;85:1359-67.
  • 10. Arnold K, Sarkar A, Yram MA, et al. Sox2(+) adult stem and progenitor cells are important for tissue regeneration and survival of mice. Cell Stem Cell. 2011;9:317-29.
  • 11. Pan GJ, Chang ZY, Schöler HR, et al. Stem cell pluripotency and transcription factor Oct4. Cell Res. 2002;12:321-329.
  • 12. Rojo de la Vega M, Chapman E, Zhang DD. NRF2 and the Hallmarks of Cancer. Cancer Cell. 2018;34:21-43.
  • 13. Menegon S, Columbano A, Giordano S. The Dual Roles of NRF2 in Cancer. Trends Mol Med. 2016;22:578-593.
  • 14. D’Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int. 2019;43:582-592.
  • 15. He F, Ru X, Wen T. NRF2, a Transcription Factor for Stress Response and Beyond. Int J Mol Sci. 2020;21:4777.
  • 16. Al-Hashimi, A.G. Antioxidant and Antibacterial Activities of Hibiscus sabdariffa L. Extracts. African Journal of Food Science. 2012;6:506-511.
  • 17. Li J, Lu YR, Lin IF, et al. Reversing UVB-induced photoaging with Hibiscus sabdariffa calyx aqueous extract. J Sci Food Agric. 2020;100:672-681.
  • 18. Malacrida A, Erriquez J, Hashemi M, et al. Evaluation of antitumoral effect of Hibiscus sabdariffa extract on human breast cancer cells. Biochem Biophys Rep. 2022;32:101353.
  • 19. Abdul Hamid Z, Lin Lin WH, Abdalla BJ, et al. The role of Hibiscus sabdariffa L. (Roselle) in maintenance of ex vivo murine bone marrow-derived hematopoietic stem cells. ScientificWorldJournal. 2014;2014:258192.
  • 20. Kaspar JW, Niture SK, Jaiswal AK. Nrf2:INrf2 (Keap1) signaling in oxidative stress. Free Radic Biol Med. 2009;47:1304-1309.
  • 21. Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem. 2009;284:13291-13295.
  • 22. Prasomthong J, Limpeanchob N, Daodee S, et al. Hibiscus sabdariffa extract improves hepatic steatosis, partially through IRS-1/Akt and Nrf2 signaling pathways in rats fed a high fat diet. Sci Rep. 2022;12:7022.
  • 23. Niture SK, Jaiswal AK. Nrf2 protein up-regulates antiapoptotic protein Bcl- 2 and prevents cellular apoptosis. J Biol Chem. 2012;287:9873-9886.
  • 24. Cengiz M, Ayhanci A, Akkemik E, et al. The role of Bax/Bcl-2 and Nrf2- Keap-1 signaling pathways in mediating the protective effect of boric acid on acrylamide-induced acute liver injury in rats. Life Sci. 2022;307:120864.
  • 25. Jaramillo MC, Zhang DD. The emerging role of the Nrf2-Keap1 signaling pathway in cancer. Genes Dev. 2013;27:2179-2191.
  • 26. Wu S, Lu H, Bai Y. Nrf2 in cancers: A double-edged sword. Cancer Med. 2019;8:2252-2267.
There are 26 citations in total.

Details

Primary Language English
Subjects Medical Biochemistry and Metabolomics (Other)
Journal Section Research Article
Authors

Elif Dener Kılıç This is me

Yüsra Nur Parlak This is me 0009-0003-5516-0829

Tuana Yıldırım This is me 0009-0001-5579-7518

Merve Çeşme This is me 0009-0000-4213-0098

Selin Akkaya This is me 0009-0000-6428-1743

Hafsa Amzım This is me 0009-0008-5654-8848

Asalat Isanbaeva This is me 0009-0001-1607-8334

Nurbanu Özgültekin This is me 0000-0002-5338-885X

Nazila Farhangzad This is me 0009-0002-3150-6103

Derya Gökmen 0000-0001-6266-3035

Tülin Özkan 0000-0002-1431-2385

Aslıhan Gürbüz Bolkan This is me 0000-0002-5089-3965

Asuman Sunguroğlu 0000-0001-7693-0958

Project Number -
Submission Date October 18, 2024
Acceptance Date October 28, 2024
Publication Date December 31, 2024
Published in Issue Year 2024 Volume: 77 Issue: 4

Cite

APA Dener Kılıç, E., Parlak, Y. N., Yıldırım, T., … Çeşme, M. (2024). Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells. Ankara Üniversitesi Tıp Fakültesi Mecmuası, 77(4), 374-380. https://doi.org/10.4274/atfm.galenos.2024.29053
AMA Dener Kılıç E, Parlak YN, Yıldırım T, et al. Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells. Ankara Üniversitesi Tıp Fakültesi Mecmuası. December 2024;77(4):374-380. doi:10.4274/atfm.galenos.2024.29053
Chicago Dener Kılıç, Elif, Yüsra Nur Parlak, Tuana Yıldırım, Merve Çeşme, Selin Akkaya, Hafsa Amzım, Asalat Isanbaeva, et al. “Anti-Aging Effects of Hibiscus Sabdariffa on Immortalized Human Ovarian Epithelial Cells”. Ankara Üniversitesi Tıp Fakültesi Mecmuası 77, no. 4 (December 2024): 374-80. https://doi.org/10.4274/atfm.galenos.2024.29053.
EndNote Dener Kılıç E, Parlak YN, Yıldırım T, Çeşme M, Akkaya S, Amzım H, Isanbaeva A, Özgültekin N, Farhangzad N, Gökmen D, Özkan T, Gürbüz Bolkan A, Sunguroğlu A (December 1, 2024) Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells. Ankara Üniversitesi Tıp Fakültesi Mecmuası 77 4 374–380.
IEEE E. Dener Kılıç et al., “Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells”, Ankara Üniversitesi Tıp Fakültesi Mecmuası, vol. 77, no. 4, pp. 374–380, 2024, doi: 10.4274/atfm.galenos.2024.29053.
ISNAD Dener Kılıç, Elif et al. “Anti-Aging Effects of Hibiscus Sabdariffa on Immortalized Human Ovarian Epithelial Cells”. Ankara Üniversitesi Tıp Fakültesi Mecmuası 77/4 (December2024), 374-380. https://doi.org/10.4274/atfm.galenos.2024.29053.
JAMA Dener Kılıç E, Parlak YN, Yıldırım T, Çeşme M, Akkaya S, Amzım H, Isanbaeva A, Özgültekin N, Farhangzad N, Gökmen D, Özkan T, Gürbüz Bolkan A, Sunguroğlu A. Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells. Ankara Üniversitesi Tıp Fakültesi Mecmuası. 2024;77:374–380.
MLA Dener Kılıç, Elif et al. “Anti-Aging Effects of Hibiscus Sabdariffa on Immortalized Human Ovarian Epithelial Cells”. Ankara Üniversitesi Tıp Fakültesi Mecmuası, vol. 77, no. 4, 2024, pp. 374-80, doi:10.4274/atfm.galenos.2024.29053.
Vancouver Dener Kılıç E, Parlak YN, Yıldırım T, Çeşme M, Akkaya S, Amzım H, et al. Anti-Aging Effects of Hibiscus sabdariffa on Immortalized Human Ovarian Epithelial Cells. Ankara Üniversitesi Tıp Fakültesi Mecmuası. 2024;77(4):374-80.