Assessment of Antimicrobial and Anti-Aging Effects of Triterpenoid and Polyphenol Compounds from Olea europaea L: Extraction, Identification and In Vitro Tests

Burçin Karabey; Ecem Saygılı; Fatih Karabey

doi:10.19161/etd.1413161

TR EN

Olea europaea L.'den elde edilen triterpenoid ve polifenol bileşiklerinin antimikrobiyal ve yaşlanma karşıtı etkilerinin değerlendirilmesi: Ekstraksiyon, Tanılama ve In Vitro Testler

Öz

Amaç: Bu çalışmada geleneksel tedavilerde Avrupa ve Akdeniz ülkelerinde kullanılmakta olan Olea europaea L. yaprakları ve çiçeklerinden elde edilen biyoaktif bileşiklerin antimikrobiyal ve yaşlanma karşıtı etkileri araştırılmıştır. Gereç ve Yöntem: Katı-sıvı ekstraksiyonu ve saflaştırma adımlarını takiben, saflaştırma işlemlerinin kontrolü ince tabaka kromatografisi (ITK) ile yapılmıştır. Elde edilen moleküllerin tanımlanması yüksek basınçlı sıvı kromatografisi (HPLC) ve nükleer manyetik rezonans (NMR) analizleri ile gerçekleştirilmiştir. Antimikrobiyal aktivitenin belirlenmesinde minimum inhibisyon konsantrasyonu (MIC) testleri için gram pozitif ve gram negatif bakterilerin temsilcisi olarak sırasıyla Bacillus cereus ve Escherichia coli O15:H7 kullanılmıştır. Oleuropein'in insan dermal fibroblastları üzerindeki in vitro sitotoksisite değerlendirmesinde, HDFa hücre hatları kullanılarak MTT testi, kollajen tip-I seviyesinde ki değişimin belirlenebilmesi için ise ELISA Test kiti kullanılmıştır. Bulgular: Çalışma sonucunda zeytin yaprağından Oleanolik asit (L1), oleouropein (L2) ve Ursolik asit (L3) molekülleri, zeytin çiçeğinden ise oleuropein aglikon (F1) molekülü izole edilerek tanımlanmıştır. Minimal inhibisyon konsantrasyonu (MIK) testi sonuçlarına göre, yapraklardan izole edilen L1, L2 ve L3 bileşikleri, Bacillus cereus'a karşı 5-250 µg / mL konsantrasyon aralığında inhibisyon etkisi göstermiştir, ancak Oleuropein aglikon (F1) herhangi bir aktivite göstermemiştir. Ayrıca, Escherichia coli'ye karşı Oleuropein (L2) molekülünün dışında hiçbir bileşiğin etkili olmadığı bulunmuştur. Oleuropein'in insan dermal fibroblastları üzerindeki in vitro değerlendirmesinde, HDFa hücrelerinin hücre canlılığı üzerinde doza bağlı bir etki gözlenmiş ve kollajen tip-I seviyeleri vitamin C ile elde edilen seviyelerden önemli ölçüde yüksek bulunmuştur. Sonuç: Elde edilen sonuçlarla olarak zeytin bitkisinden elde edilen aktif moleküllerin antimikrobiyal etkileri ile doğal koruyucu olarak kozmetik alanında ve hücre yenilenmesine olan katkısı ile yara tedavilerinde potansiyel kullanımı olduğu düşünülmektedir.

Anahtar Kelimeler

Assessment of Antimicrobial and Anti-Aging Effects of Triterpenoid and Polyphenol Compounds from Olea europaea L: Extraction, Identification and In Vitro Tests

Abstract

Aim: This study investigated the antimicrobial and anti-aging effects of bioactive compounds derived from Olea europaea L. leaves and flowers, widely used in traditional treatments in European and Mediterranean countries. Materials and Methods: Following solid-liquid extraction and purification steps, the control of purification processes was conducted using thin-layer chromatography (TLC). Identification of the obtained molecules was performed through high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) analyses. For determining antimicrobial activity, representatives of Gram-positive and Gram-negative bacteria, Bacillus cereus, and Escherichia coli O15:H7, were respectively used for minimum inhibitory concentration (MIC) tests. In the assessment of Oleuropein's in vitro cytotoxicity on human dermal fibroblasts, the MTT assay was employed using HDFa cell lines, while an ELISA Test kit was utilized to determine changes in collagen type-I levels. Results: As a result of the study, molecules oleanolic acid (L1), oleuropein (L2), and ursolic acid (L3) were isolated and identified from olive leaves, whereas oleuropein aglycone (F1) molecule was isolated from olive flowers. According to the results of the Minimum Inhibitory Concentration (MIC) tests, compounds L1, L2, and L3 isolated from the leaves exhibited an inhibitory effect against Bacillus cereus within the concentration range of 5-250 µg/mL, whereas Oleuropein aglycone (F1) did not demonstrate any inhibitory activity. Furthermore, it was found that, except for the Oleuropein (L2) molecule, no other compound was effective against Escherichia coli. In the evaluation of Oleuropein's in vitro cytotoxicity on human dermal fibroblasts, a dose-dependent effect on HDFa cell viability was observed, and collagen type-I levels were significantly higher than levels obtained with vitamin C. Conclusion: Based on the obtained results, it is believed that the active molecules derived from olive plant's leaves and flowers exhibit antimicrobial effects, potentially serving as natural preservatives in the cosmetics industry. Moreover, their contribution to cell regeneration suggests potential use in wound treatments

Keywords

References

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Details

Primary Language

English

Subjects

Clinical Microbiology

Journal Section

Research Article

Authors

Burçin Karabey ^*
0000-0003-4834-6740
Türkiye

Ecem Saygılı
0000-0002-8389-9079
Türkiye

Fatih Karabey
0000-0003-0305-6192
Türkiye

Publication Date

September 9, 2024

Submission Date

January 10, 2024

Acceptance Date

February 13, 2024

Published in Issue

Year 2024 Volume: 63 Number: 3

DOI

https://doi.org/10.19161/etd.1413161

IZ

https://izlik.org/JA63SW52AP

Cite

RIS / Bibtex

Vancouver

1.Burçin Karabey, Ecem Saygılı, Fatih Karabey. Assessment of Antimicrobial and Anti-Aging Effects of Triterpenoid and Polyphenol Compounds from Olea europaea L: Extraction, Identification and In Vitro Tests. EJM. 2024 Sep. 1;63(3):369-77. doi:10.19161/etd.1413161