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
- 1. Borges A, José H, Homem V and Simões M. Comparison of techniques and solvents on the antimicrobial and antioxidant potential of extracts from Acacia dealbata and Olea europaea. Antibiotics.2020; 9: 48.
- 2. Ergulen E, Ozkaya MT, Ulger S aND Ozilbey N. Identification of some Turkish olive cultivars by using RAPD-PCR technique. Acta Hortic.2002; 586: 91-5.
- 3. Davis PH. Flora of Turkey and the East Aegean Islands Vol.3. Edinburgh. Edinburgh University Press;1975.
- 4. Putnik P, Lorenzo J, Barba F, Roohinejad S, Režek Jambrak A, Granato D, Montesano D and Bursa´c Kovaˇcevi´c D. Novel food processing and extraction technologies of high-added value compounds from plant materials. Foods.2018; 7: 106.
- 5. Giacometti J, Bursa´c Kovaˇcevi´c D, Putnik P, Gabri´c D, Biluši´c T, Kreši´c G, Stuli´c V, Barba FJ, Chemat F and Barbosa-Cánovas G. Extraction of bioactive compounds and essential oils from mediterranean herbs by conventional and green innovative techniques: A review. Food Res. Int. 2018; 113: 245–62.
- 6. Ferreira ICFR, Barros L, Soares ME, Bastos ML and Pereira JA. Antioxidant activity and phenolic contents of Olea europaea l. leaves sprayed with different copper formulations. Food Chem. 2007; 103: 188–95.
- 7. Jung YC, Kim HW, Min BK, Cho JY, Son HJ, Lee JY, Kim JY, Kwon SB, Li Q and Lee HW. Inhibitory effect of olive leaf extract on obesity in high-fat diet-induced mice. In Vivo.2019; 33: 707–15.
- 8. Batçıoğlu K; Küçükbay FZ, Alagöz MA, Günal S and Yilmaztekin Y. Antioxidant and antithrombotic properties of fruit, leaf, and seed extracts of the Halhalı olive (Olea europaea L.) native to the Hatay region in Turkey. Food Raw Mater. 2023; 11: 84–93.
- 9. Sun W, Frost B and Liu J. Oleuropein, Unexpected Benefits! Oncotarget 2017; 8:17409.
- 10. Yu H, Liu P, Tang H, Jing J, Lv X, Chen L, Jiang L, Xu J and Li J. Oleuropein, a natural extract from plants, offers neuroprotection in focal cerebral ischemia/reperfusion injury in mice. Eur. J. Pharmacol.2016; 775: 113–9.
- 11. Antoniou C and Hull J. The anti-cancer effect of Olea europaea L. products: a Review. Cur Nutr Rep. 2021; 10: 99–124.
- 12. Ćujić N, Šavikin K, Janković T, Pljevljakušić D, Zdunić G and Ibrić S. Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique. Food Chem. 2015; 194:135 42.
- 13. Recepoglu YK, Gümüşoğlu G and Özşen AY. Comparative assessment for efficient oleuropein extraction from olive leaf (Olea europaea L. folium). Turkish Journal of Engineering. 2023; 7(2): 116-24.
- 14. National Committee for Clinical Laboratory Standards (2001) Performance standards for antimicrobial susceptibility testing; 11th informational supplement. M100-S11. National Committee for Clinical Laboratory Standards, Wayne, Pa.
- 15. Topuz S and Bayram M. Oleuropein extraction from leaves of three olive varieties (Olea europaea L.): Antioxidant and antimicrobial properties of purified oleuropein and oleuropein extracts. Journal of Food Processing and Preservation. 2021; 46:15697.
- 16. Palmeri R, Parafati L, Trippa D, Siracusa L, Arena E, Restuccia C and Fallico B. Addition of olive leaf extract (OLE) for producing fortified fresh pasteurized milk with an extended shelf life. Antioxidants.2019; 8: 255.
- 17. Asgharzade S, Sheikhshabani SH, Ghasempour E, Heidari R, Rahmati S, Mohammadi M, Jazaeri A and Amini-Farsani Z. The effect of oleuropein on apoptotic pathway regulators in breast cancer cells. European Journal of Pharmacology.2020; 886: 173509.
- 18. Sherif IO and Al-Gayyar MMH. Oleuropein potentiates anti-tumor activity of cisplatin against HepG2 through affecting proNGF/NGF balance. Life Sciences.2018; 198: 87–93.
- 19. Morandi F, Bensa V, Calarco E, Pastorino F, Perri P, Corrias MV, Ponzoni M and Brignole C. The olive leaves extract has anti‐tumor effects against neuroblastoma through inhibition of cell proliferation and induction of apoptosis. Nutrients. 2021; 13:7.
- 20. Bal Y, Sürmeli Y and Şanlı-Mohamed G. Antiproliferative and apoptotic effects of olive leaf extract microcapsules on MCF-7 and A549 cancer cells. ACS Omega.2023;8(32): 28984–28993.
- 21. Chotphruethipong L, Hutamekalin P, Nilsuwan K, Sukketsiri W, Aluko RE, Abdul NR and Benjakul S. Combined effects of defatted hydrolyzed collagen from salmon skin and vitamin C on proliferation and migration of human fibroblast cell. Fishes. 2022; 7(5): 265.
- 22. Munira SRYWW. The effect of vitamin C on fibroblast proliferation and VEGF expression in fibroblast culture. Journal of the Medical Sciences (Berkala Ilmu Kedokteran).2015; 41(03): 152-6.
- 23. Goldsmith CD, Bond DR, Jankowski H, Weidenhofer J, Stathopoulos CE, Roach PD and Scarlett CJ. The olive biophenols oleuropein and hydroxytyrosol selectively reduce proliferation, influence the cell cycle, and induce apoptosis in pancreatic cancer cells. International Journal of Molecular Sciences.2018; 19(7): 1937.
- 24. Katsiki M, Chondrogianni N, Chinou I, Rivett AJ and Gonos ES. The olive constituent oleuropein exhibits proteasome stimulatory properties in vitro and confers life span extension of human embryonic fibroblasts. Rejuvenation Res. 2007; 10(2): 157-72.
- 25. Oliveira ALS, Gondim S, Gómez-García R, Ribeiro T and Pintado M. Olive leaf phenolic extract from two Portuguese cultivars – bioactivities for potential food and cosmetic application. Journal of Environmental Chemical Engineering.2021; 9(5):106175.
- 26. Mehraein F, Sarbishegi M and Aslani M. Evaluation of effect of oleuropein on skin wound healing in aged male Balb/c mice. Cell Journal (Yakhteh).2014; 16(1): 25.
Olea europaea L.'den elde edilen triterpenoid ve polifenol bileşiklerinin antimikrobiyal ve yaşlanma karşıtı etkilerinin değerlendirilmesi: Ekstraksiyon, Tanılama ve In Vitro Testler
Abstract
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.
References
- 1. Borges A, José H, Homem V and Simões M. Comparison of techniques and solvents on the antimicrobial and antioxidant potential of extracts from Acacia dealbata and Olea europaea. Antibiotics.2020; 9: 48.
- 2. Ergulen E, Ozkaya MT, Ulger S aND Ozilbey N. Identification of some Turkish olive cultivars by using RAPD-PCR technique. Acta Hortic.2002; 586: 91-5.
- 3. Davis PH. Flora of Turkey and the East Aegean Islands Vol.3. Edinburgh. Edinburgh University Press;1975.
- 4. Putnik P, Lorenzo J, Barba F, Roohinejad S, Režek Jambrak A, Granato D, Montesano D and Bursa´c Kovaˇcevi´c D. Novel food processing and extraction technologies of high-added value compounds from plant materials. Foods.2018; 7: 106.
- 5. Giacometti J, Bursa´c Kovaˇcevi´c D, Putnik P, Gabri´c D, Biluši´c T, Kreši´c G, Stuli´c V, Barba FJ, Chemat F and Barbosa-Cánovas G. Extraction of bioactive compounds and essential oils from mediterranean herbs by conventional and green innovative techniques: A review. Food Res. Int. 2018; 113: 245–62.
- 6. Ferreira ICFR, Barros L, Soares ME, Bastos ML and Pereira JA. Antioxidant activity and phenolic contents of Olea europaea l. leaves sprayed with different copper formulations. Food Chem. 2007; 103: 188–95.
- 7. Jung YC, Kim HW, Min BK, Cho JY, Son HJ, Lee JY, Kim JY, Kwon SB, Li Q and Lee HW. Inhibitory effect of olive leaf extract on obesity in high-fat diet-induced mice. In Vivo.2019; 33: 707–15.
- 8. Batçıoğlu K; Küçükbay FZ, Alagöz MA, Günal S and Yilmaztekin Y. Antioxidant and antithrombotic properties of fruit, leaf, and seed extracts of the Halhalı olive (Olea europaea L.) native to the Hatay region in Turkey. Food Raw Mater. 2023; 11: 84–93.
- 9. Sun W, Frost B and Liu J. Oleuropein, Unexpected Benefits! Oncotarget 2017; 8:17409.
- 10. Yu H, Liu P, Tang H, Jing J, Lv X, Chen L, Jiang L, Xu J and Li J. Oleuropein, a natural extract from plants, offers neuroprotection in focal cerebral ischemia/reperfusion injury in mice. Eur. J. Pharmacol.2016; 775: 113–9.
- 11. Antoniou C and Hull J. The anti-cancer effect of Olea europaea L. products: a Review. Cur Nutr Rep. 2021; 10: 99–124.
- 12. Ćujić N, Šavikin K, Janković T, Pljevljakušić D, Zdunić G and Ibrić S. Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique. Food Chem. 2015; 194:135 42.
- 13. Recepoglu YK, Gümüşoğlu G and Özşen AY. Comparative assessment for efficient oleuropein extraction from olive leaf (Olea europaea L. folium). Turkish Journal of Engineering. 2023; 7(2): 116-24.
- 14. National Committee for Clinical Laboratory Standards (2001) Performance standards for antimicrobial susceptibility testing; 11th informational supplement. M100-S11. National Committee for Clinical Laboratory Standards, Wayne, Pa.
- 15. Topuz S and Bayram M. Oleuropein extraction from leaves of three olive varieties (Olea europaea L.): Antioxidant and antimicrobial properties of purified oleuropein and oleuropein extracts. Journal of Food Processing and Preservation. 2021; 46:15697.
- 16. Palmeri R, Parafati L, Trippa D, Siracusa L, Arena E, Restuccia C and Fallico B. Addition of olive leaf extract (OLE) for producing fortified fresh pasteurized milk with an extended shelf life. Antioxidants.2019; 8: 255.
- 17. Asgharzade S, Sheikhshabani SH, Ghasempour E, Heidari R, Rahmati S, Mohammadi M, Jazaeri A and Amini-Farsani Z. The effect of oleuropein on apoptotic pathway regulators in breast cancer cells. European Journal of Pharmacology.2020; 886: 173509.
- 18. Sherif IO and Al-Gayyar MMH. Oleuropein potentiates anti-tumor activity of cisplatin against HepG2 through affecting proNGF/NGF balance. Life Sciences.2018; 198: 87–93.
- 19. Morandi F, Bensa V, Calarco E, Pastorino F, Perri P, Corrias MV, Ponzoni M and Brignole C. The olive leaves extract has anti‐tumor effects against neuroblastoma through inhibition of cell proliferation and induction of apoptosis. Nutrients. 2021; 13:7.
- 20. Bal Y, Sürmeli Y and Şanlı-Mohamed G. Antiproliferative and apoptotic effects of olive leaf extract microcapsules on MCF-7 and A549 cancer cells. ACS Omega.2023;8(32): 28984–28993.
- 21. Chotphruethipong L, Hutamekalin P, Nilsuwan K, Sukketsiri W, Aluko RE, Abdul NR and Benjakul S. Combined effects of defatted hydrolyzed collagen from salmon skin and vitamin C on proliferation and migration of human fibroblast cell. Fishes. 2022; 7(5): 265.
- 22. Munira SRYWW. The effect of vitamin C on fibroblast proliferation and VEGF expression in fibroblast culture. Journal of the Medical Sciences (Berkala Ilmu Kedokteran).2015; 41(03): 152-6.
- 23. Goldsmith CD, Bond DR, Jankowski H, Weidenhofer J, Stathopoulos CE, Roach PD and Scarlett CJ. The olive biophenols oleuropein and hydroxytyrosol selectively reduce proliferation, influence the cell cycle, and induce apoptosis in pancreatic cancer cells. International Journal of Molecular Sciences.2018; 19(7): 1937.
- 24. Katsiki M, Chondrogianni N, Chinou I, Rivett AJ and Gonos ES. The olive constituent oleuropein exhibits proteasome stimulatory properties in vitro and confers life span extension of human embryonic fibroblasts. Rejuvenation Res. 2007; 10(2): 157-72.
- 25. Oliveira ALS, Gondim S, Gómez-García R, Ribeiro T and Pintado M. Olive leaf phenolic extract from two Portuguese cultivars – bioactivities for potential food and cosmetic application. Journal of Environmental Chemical Engineering.2021; 9(5):106175.
- 26. Mehraein F, Sarbishegi M and Aslani M. Evaluation of effect of oleuropein on skin wound healing in aged male Balb/c mice. Cell Journal (Yakhteh).2014; 16(1): 25.
Details
| Primary Language | English |
|---|---|
| Subjects | Clinical Microbiology |
| Journal Section | Research Articles |
| Authors | |
| Publication Date | September 9, 2024 |
| Submission Date | January 10, 2024 |
| Acceptance Date | February 13, 2024 |
| Published in Issue | Year 2024 |