CISTUS CRETICUS L.: ANTIOBESITY, ANTIMICROBIAL AND ANTIBIOFILM PROPERTIES
Year 2025,
Volume: 49 Issue: 1, 119 - 132
Asli Can Agca
,
Derya Altay
,
Başar Karaca
,
Sibel Ozdas
,
İpek Canatar
Abstract
Objective: Cistus creticus L. is widespread in the coastal regions of Türkiye. In this study, we investigated the cytotoxic, antiobesity, antimicrobial and antibiofilm properties as well as the total phenolic and flavonoid content of both aqueous and ethanolic extracts in vitro.
Material and Method: Two different extracts were prepared from the flowering aerial parts of Cistus creticus using ethanol and water. The total phenolic content and total flavonoids were determined by the Folin-Ciocalteu method and the aluminum chloride colorimetry, respectively. The effect of extracts on the cell viability of 3T3-L1 was determined by methyl thiazole tetrazolium (MTT), and the evaluation of differentiation and the effects of the plant extracts on lipid accumulation in 3T3-L1 adipocytes was performed by Oil-Red O staining. In addition, MIC values and antibiofilm activities were also investigated.
Result and Discussion: The total phenol content of the EtOH and water extract was determined to be 134.2849 mg GAE/g and 96.1803 mg GAE/g, respectively. The total flavonoids in the water and EtOH extracts were found to be 33.1942 mgQE/g and 22.8338 mgQE/g, respectively. The lowest MIC values were determined for the strains Bacillus subtilis DSM 1971, Bacillus licheniformis DSM 13 and Bacillus amyloliquefaciens DSM 7, while the highest MIC concentration was found for the strains Escherichia coli and Eenterococcus gallinarum. The MIC/16 concentration of Pseudomonas aeruginosa ATCC 27853 and Escherichia coli ATCC 25922 also proved to be effective in inhibiting biofilm formation. We observed that noticeable but not strong effects on lipid accumulation were observed in 3T3-L1 adipocytes treated with EtOH extract.
Ethical Statement
The authors declare that the ethics committee approval is not required for this study.
Thanks
The authors would like to thank Prof. Dr. Hayri Duman (Gazi University, Faculty of Science, Department of Biology) for the identification of the plant material.
References
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- 9. Watnick, P., Kolter, R. (2000). Biofilm, city of microbes. Journal of Bacteriology, 182(10), 2675-2679. [CrossRef]
- 10. Lewis, K. (2001). Riddle of biofilm resistance. Antimicrobial Agents and Chemotherapy, 45(4), 999-1007. [CrossRef]
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- 12. Erdoğmuş, S.F., Bilecen, C., Erdal-Altıntaş, Ö., Ulukütük, S., Kargıoğlu, M. (2022). The antibiofilm effects of some Cistus spp. against pathogenic microorganisms. International Journal of Plant Based Pharmaceuticals, 2(2), 252-260. [CrossRef]
- 13. Onal, N.F., Ozturk, I., Aydin Kose, F., Der, G., Kilinc, E., Baykan, S. (2023). Comparative evaluation of polyphenol contents and biological activities of five Cistus L. species native to Turkey. Chemistry & Biodiversity, 20(1), e202200915. [CrossRef]
- 14. Mocan, A., Fernandes, Â., Calhelha, R. C., Gavrilaş, L., Ferreira, I.C., Ivanov, M., Sokovic, M., Barros, L., Babotă, M. (2022). Bioactive compounds and functional properties of herbal preparations of Cystus creticus L. collected from Rhodes Island. Frontiers in Nutrition, 9, 881210. [CrossRef]
- 15. Hickl, J., Argyropoulou, A., Sakavitsi, M. E., Halabalaki, M., Al-Ahmad, A., Hellwig, E., Aligiannis N., Skaltssounis, A.L., Wittmer, A., Vach, K., Karygianni, L. (2018). Mediterranean herb extracts inhibit microbial growth of representative oral microorganisms and biofilm formation of Streptococcus mutans. PLoS One, 13(12), e0207574. [CrossRef]
- 16. Singleton, V.L., Rossi, J.A. (1965). Colorimetry of total phenolics width phosphomolybdic phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144-158. [CrossRef]
- 17. Zhinsen, J., Mengcheng, T., Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry, 64(4), 555-559. [CrossRef]
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- 19. Römling, U., Sierralta, W.D., Eriksson, K., Normark, S. (1998). Multicellular and aggregative behavior of Salmonella typhimurium strains is controlled by mutations in the agfD promoter. Molecular Microbiology, 28(2), 249-264. [CrossRef]
- 20. Stepanović, S., Vuković, D., Dakić, I., Savić, B., Švabić-Vlahović, M. (2000). A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods, 40(2), 175-179. [CrossRef]
- 21. Vestby, L.K., Møretrø, T., Langsrud, S., Heir, E., Nesse, L.L. (2009). Biofilm-forming abilities of Salmonella are correlated with persistence in fish meal and feed factories. BMC Veterinary Research, 5(20), 1-6. [CrossRef]
- 22. Onbas, T., Osmanagaoglu, O., Kiran, F. (2019). Potential properties of Lactobacillus plantarum F-10 as a bio-control strategy for wound infections. Probiotics and Antimicrobial Proteins, 11, 1110-1123. [CrossRef]
- 23. Longo, M., Spinelli, R., D’Esposito, V., Zatterale, F., Fiory, F., Nigro, C., Raciti, G.A., Miele, C., Formisano, P., Beguinot, F., Di Jaso, B. (2016). Pathologic endoplasmic reticulum stress induced by glucotoxic insults inhibits adipocyte differentiation and induces an inflammatory phenotype. Biochimica et Biophysica Acta, 1863(6), 1146-1156. [CrossRef]
- 24. Lee, H.W., Rhee, D.K., Kim, B.O., Pyo, S. (2018). Inhibitory effect of sinigrin on adipocyte differentiation in 3T3-L1 cells: Involvement of AMPK and MAPK pathways. Biomedicine and Pharmacotherapy, 102, 670-680. [CrossRef]
- 25. Ho, J.N., Son, M.E., Lim, W.C., Lim, S.T., Cho, H.Y. (2012). Anti-obesity effects of germinated brown rice extract through down-regulation of lipogenic genes in high fat diet-induced obese mice. Bioscience, Biotechnology and Biochemistry, 76(6), 1068-1074. [CrossRef]
- 26. Raciti, G.A., Fiory, F., Campitelli, M., Desiderio, A., Spinelli, R., Longo, M., Nigro, C., Pepe, G., Sommella, E., Campiglia, P., Formisano, P., Beguinot, F., Miele, C. (2018). Citrus aurantium L. dry extracts promote C/ebpβ expression and improve adipocyte differentiation in 3T3-L1 cells. PLoS One, 13(3), 0193704. [CrossRef]
- 27. Karunakaran, R.S., Lokanatha, O., Muni-Swamy, G., Venkataramaiah, C., Muni-Kesavulu, M., Appa-Rao, C., Badri, K.R., Balaji M. (2021). Anti-obesity and lipid-lowering activity of Bauhiniastatin-1 is mediated through PPAR-γ/AMPK expressions in diet-induced obese rat model. Frontiers in Pharmacology, 12, 704074. [CrossRef]
- 28. Liu, X.F., Bera, T.K., Kahue, C., Escobar, T., Fei, Z., Raciti, G.A., Pastan, I. (2012). ANKRD26 and its interacting partners TRIO, GPS2, HMMR and DIPA regulate adipogenesis in 3T3-L1 cells. PLoS One, 7(5), e38130. [CrossRef]
- 29. Nunez, S.K., Young, C., Adebayo, O., Muppuru, K.M., Badri, K.R. (2019). P311, a novel intrinsically disordered protein, regulates adipocyte development. Biochemical and Biophysical Research Communications, 515(1), 234-240. [CrossRef]
- 30. Lu, T., Xu, Y., Mericle, M.T., Mellgren, R.L. (2002). Participation of the conventional calpains in apoptosis. Biochimica et Biophysica Acta, 1590(1-3), 16-26. [CrossRef]
- 31. Per, S., Ercan, F.S. (2015). Comparison of three methods of DNA extraction from Parachipteria willmanni (Acari: Oribatida) collected in Turkey. Journal of Biotechnology Research, 1(4), 16-20.
- 32. Glasel, J.A. (1995). Validity of nucleic acid purities monitored by A260/A280 absorbance ratios. Biotechniques,18(1), 62-63.
- 33. Manchester, K.L. (1996). Use of UV methods for measurement of protein and nucleic acid concentrations. Biotechniques, 20(6), 968-970. [CrossRef]
- 34. Burton, K. (1956). The study of the conditions and mechanisms of the diphenylamine reaction for the calorimetric estimation of deoxyribonucleic acid. The Biochemical Journal, 62(2), 615-623. [CrossRef]
- 35. Gedikoğlu, A., Öztürk, H.İ., Aytaç, E. (2023). The effect of different solvents on chemical composition, antioxidant activity, and antimicrobial potential of Turkish Cistus creticus extracts. GIDA, 48(4), 728-740. [CrossRef]
- 36. Lukas, B., Bragagna, L., Starzyk, K., Labedz, K., Stolze, K., Novak, J. (2021). Polyphenol diversity and antioxidant activity of European Cistus creticus L. (Cistaceae) compared to six further, partly sympatric Cistus species. Plants, 10(4), 615. [CrossRef]
- 37. Demetzos, C., Loukis, A., Spiliotis, V., Zoakis, N., Stratigakis, N., Katerinopoulos, H.E. (1995). Composition and antimicrobial activity of the essential oil of Cistus creticus L. Journal of Essential Oil Research, 7(4), 407-410. [CrossRef]
- 38. Demetzos, C., Stahl, B., Anastassaki, T., Gazouli, M., Tzouvelekis, L.S., Rallis, M. (1999). Chemical analysis and antimicrobial activity of the Resin Ladano, of its essential oil and of the isolated compounds. Planta Medica, 65(1), 76-78. [CrossRef]
- 39. Güvenç¸A., Yıldız, S., Özkan, A.M., Erdurak, C.S., Coşkun, M., Yılmaz, G., Okuyama, T., Okada (2005). Antimicrobiological studies on Turkish Cistus species. Pharmaceutical Biology 43(2), 178-183. [CrossRef]
- 40. Arıtuluk, Z.C. (2010). Master of Science Thesis. Flora and folk medicines of Tefenni (Burdur) District. Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
- 41. Sargin, S.A. (2021). Plants used against obesity in Turkish folk medicine: A review. Journal of Ethnopharmacology, 270, 113841. [CrossRef]
CİSTUS CRETİCUS L.: ANTİOBEZİTE, ANTİMİKROBİYAL VE ANTİBİYOFİLM ÖZELLİKLERİ
Year 2025,
Volume: 49 Issue: 1, 119 - 132
Asli Can Agca
,
Derya Altay
,
Başar Karaca
,
Sibel Ozdas
,
İpek Canatar
Abstract
Amaç: Cistus creticus L. Türkiye'nin kıyı bölgelerinde yaygın olarak bulunur. Bu çalışmada, sulu ve etanollü ekstrelerin sitotoksik, antiobezite, antimikrobiyal ve antibiyofilm özelliklerinin yanı sıra toplam fenolik ve flavonoit içeriği in vitro olarak araştırılmıştır.
Gereç ve Yöntem: Cistus creticus'un çiçekli toprak üstü kısımlarından hareketle etanol ve su kullanılarak iki farklı ekstre hazırlanmıştır. Toplam fenolik içerik ve toplam flavonoitler sırasıyla Folin-Ciocalteu ve alüminyum klorür kolorimetrik yöntemleriyle belirlenmiştir. Cistus creticus'un toprak üstü kısımlarının etanollü ve sulu ekstrelerinin 3T3-L1 hücre canlılığı üzerindeki etkisi metil tiyazol tetrazolyum (MTT) ile belirlenmiş ve bitki ekstrelerinin 3T3-L1 adipositlerinde farklılaşma ve lipit birikimi üzerindeki etkileri Yağ-Kırmızı O Boyama ile değerlendirilmiştir. Ayrıca, MIC değerleri ve antibiyofilm aktiviteleri de incelenmiştir.
Sonuç ve Tartışma: EtOH ve sulu ekstrelerin toplam fenol içeriği sırasıyla 134.2849 mg GAE/g ve 96,1803 mg GAE/g olarak belirlenmiştir. Su ve EtOH ekstrelerindeki toplam flavonoitlerin sırasıyla 33.1942 mgKE/g ve 22.8338 mgKE/g olduğu bulunmuştur. En düşük MİK değerleri Bacillus subtilis DSM 1971, Bacillus licheniformis DSM 13 ve Bacillus amyloliquefaciens DSM 7 suşları için belirlenirken, en yüksek MİK konsantrasyonu Escherichia coli ve Enteroccus gallinarum suşları için bulunmuştur. Pseudomonas aeruginosa ATCC 27853 ve Escherichia coli ATCC 25922'nin MIC/16 konsantrasyonunun da biyofilm oluşumunu engellemede etkili olduğu kanıtlanmıştır. EtOH ekstresi ile muamele edilen 3T3-L1 adipositlerinde lipid birikimi üzerinde belirgin ancak güçlü olmayan etkiler gözlemledik.
References
- 1. Tanker, N., Koyuncu, M., Coşkun, M. (2007). Farmasötik Botanik, Ankara Üniversitesi Basımevi, Ankara, p. 245.
- 2. Baytop T. (1999). Türkiye’de Bitkilerle Tedavi, Nobel Tıp Kitapevleri, İstanbul, p. 284.
- 3. Tuzlacı E. (2016). Türkiye Bitkileri Geleneksel İlaç Rehberi, İstanbul Medika Yayıncılık, İstanbul, p. 53, 311, 667-668, 822.
- 4. Barrajón-Catalán, E., Fernández-Arroyo, S., Roldán, C., Guillén, E., Saura, D., Segura-Carreteroc, A., Micola, V. (2011). A systematic study of the polyphenolic composition of aqueous extracts deriving from several Cistus genus species: Evolutionary relationship. Phytochemical Analysis, 22, 303-312. [CrossRef]
- 5. Tomou, E-M., Krystalia-Lytra, K., Rallis, S., Tzakos, A.G., Skaltsa, H. (2022). An updated review of genus Cistus L. since 2014: Traditional uses, phytochemistry, and pharmacological properties. Phytochemical Review, 21, 2049-2087. [CrossRef]
- 6. El-Ghazouani, F., El-Ouahmani, N., Teixidor-Toneu, I, Yacoubi, B., Zekhnini, A. (2021). A survey of medicinal plants used in traditional medicine by women and herbalists from the city of Agadir, southwest of Morocco. European Journal of Integrative Medicine, 42, 101284. [CrossRef]
- 7. World Health Organisation Web site. (2024). Retrieved date March 1, 2024, from https://www.who.int/. Accessed date: 05.03.2024.
- 8. Zalegh, I., Akssira, M., Bourhia, M., Mellouki, F., Rhallabi, N., Salamatullah, A.M., Alkaltham, M.S., Alyahya, H.K., Mhand, R.A. (2021). A review on Cistus sp.: Phytochemical and antimicrobial activities. Plants, 10(6), 1214. [CrossRef]
- 9. Watnick, P., Kolter, R. (2000). Biofilm, city of microbes. Journal of Bacteriology, 182(10), 2675-2679. [CrossRef]
- 10. Lewis, K. (2001). Riddle of biofilm resistance. Antimicrobial Agents and Chemotherapy, 45(4), 999-1007. [CrossRef]
- 11. Lu, L., Hu, W., Tian, Z., Yuan, D., Yi, G., Zhou, Y., Cheng, Q., Zhu, J., Li, M. (2019). Developing natural products as potential anti-biofilm agents. Chinese Medicine, 14(11), 1-17. [CrossRef]
- 12. Erdoğmuş, S.F., Bilecen, C., Erdal-Altıntaş, Ö., Ulukütük, S., Kargıoğlu, M. (2022). The antibiofilm effects of some Cistus spp. against pathogenic microorganisms. International Journal of Plant Based Pharmaceuticals, 2(2), 252-260. [CrossRef]
- 13. Onal, N.F., Ozturk, I., Aydin Kose, F., Der, G., Kilinc, E., Baykan, S. (2023). Comparative evaluation of polyphenol contents and biological activities of five Cistus L. species native to Turkey. Chemistry & Biodiversity, 20(1), e202200915. [CrossRef]
- 14. Mocan, A., Fernandes, Â., Calhelha, R. C., Gavrilaş, L., Ferreira, I.C., Ivanov, M., Sokovic, M., Barros, L., Babotă, M. (2022). Bioactive compounds and functional properties of herbal preparations of Cystus creticus L. collected from Rhodes Island. Frontiers in Nutrition, 9, 881210. [CrossRef]
- 15. Hickl, J., Argyropoulou, A., Sakavitsi, M. E., Halabalaki, M., Al-Ahmad, A., Hellwig, E., Aligiannis N., Skaltssounis, A.L., Wittmer, A., Vach, K., Karygianni, L. (2018). Mediterranean herb extracts inhibit microbial growth of representative oral microorganisms and biofilm formation of Streptococcus mutans. PLoS One, 13(12), e0207574. [CrossRef]
- 16. Singleton, V.L., Rossi, J.A. (1965). Colorimetry of total phenolics width phosphomolybdic phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144-158. [CrossRef]
- 17. Zhinsen, J., Mengcheng, T., Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry, 64(4), 555-559. [CrossRef]
- 18. Clinical and Laboratory Standards Institute (CLSI) Web site. (2024). M100-S25 performance standards for antimicrobial susceptibility testing; Twenty-fifth informational supplement, USA. Retrieved date March 1, 2024, from https://www.who.int/. Accessed date: 05.03.2024.
- 19. Römling, U., Sierralta, W.D., Eriksson, K., Normark, S. (1998). Multicellular and aggregative behavior of Salmonella typhimurium strains is controlled by mutations in the agfD promoter. Molecular Microbiology, 28(2), 249-264. [CrossRef]
- 20. Stepanović, S., Vuković, D., Dakić, I., Savić, B., Švabić-Vlahović, M. (2000). A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods, 40(2), 175-179. [CrossRef]
- 21. Vestby, L.K., Møretrø, T., Langsrud, S., Heir, E., Nesse, L.L. (2009). Biofilm-forming abilities of Salmonella are correlated with persistence in fish meal and feed factories. BMC Veterinary Research, 5(20), 1-6. [CrossRef]
- 22. Onbas, T., Osmanagaoglu, O., Kiran, F. (2019). Potential properties of Lactobacillus plantarum F-10 as a bio-control strategy for wound infections. Probiotics and Antimicrobial Proteins, 11, 1110-1123. [CrossRef]
- 23. Longo, M., Spinelli, R., D’Esposito, V., Zatterale, F., Fiory, F., Nigro, C., Raciti, G.A., Miele, C., Formisano, P., Beguinot, F., Di Jaso, B. (2016). Pathologic endoplasmic reticulum stress induced by glucotoxic insults inhibits adipocyte differentiation and induces an inflammatory phenotype. Biochimica et Biophysica Acta, 1863(6), 1146-1156. [CrossRef]
- 24. Lee, H.W., Rhee, D.K., Kim, B.O., Pyo, S. (2018). Inhibitory effect of sinigrin on adipocyte differentiation in 3T3-L1 cells: Involvement of AMPK and MAPK pathways. Biomedicine and Pharmacotherapy, 102, 670-680. [CrossRef]
- 25. Ho, J.N., Son, M.E., Lim, W.C., Lim, S.T., Cho, H.Y. (2012). Anti-obesity effects of germinated brown rice extract through down-regulation of lipogenic genes in high fat diet-induced obese mice. Bioscience, Biotechnology and Biochemistry, 76(6), 1068-1074. [CrossRef]
- 26. Raciti, G.A., Fiory, F., Campitelli, M., Desiderio, A., Spinelli, R., Longo, M., Nigro, C., Pepe, G., Sommella, E., Campiglia, P., Formisano, P., Beguinot, F., Miele, C. (2018). Citrus aurantium L. dry extracts promote C/ebpβ expression and improve adipocyte differentiation in 3T3-L1 cells. PLoS One, 13(3), 0193704. [CrossRef]
- 27. Karunakaran, R.S., Lokanatha, O., Muni-Swamy, G., Venkataramaiah, C., Muni-Kesavulu, M., Appa-Rao, C., Badri, K.R., Balaji M. (2021). Anti-obesity and lipid-lowering activity of Bauhiniastatin-1 is mediated through PPAR-γ/AMPK expressions in diet-induced obese rat model. Frontiers in Pharmacology, 12, 704074. [CrossRef]
- 28. Liu, X.F., Bera, T.K., Kahue, C., Escobar, T., Fei, Z., Raciti, G.A., Pastan, I. (2012). ANKRD26 and its interacting partners TRIO, GPS2, HMMR and DIPA regulate adipogenesis in 3T3-L1 cells. PLoS One, 7(5), e38130. [CrossRef]
- 29. Nunez, S.K., Young, C., Adebayo, O., Muppuru, K.M., Badri, K.R. (2019). P311, a novel intrinsically disordered protein, regulates adipocyte development. Biochemical and Biophysical Research Communications, 515(1), 234-240. [CrossRef]
- 30. Lu, T., Xu, Y., Mericle, M.T., Mellgren, R.L. (2002). Participation of the conventional calpains in apoptosis. Biochimica et Biophysica Acta, 1590(1-3), 16-26. [CrossRef]
- 31. Per, S., Ercan, F.S. (2015). Comparison of three methods of DNA extraction from Parachipteria willmanni (Acari: Oribatida) collected in Turkey. Journal of Biotechnology Research, 1(4), 16-20.
- 32. Glasel, J.A. (1995). Validity of nucleic acid purities monitored by A260/A280 absorbance ratios. Biotechniques,18(1), 62-63.
- 33. Manchester, K.L. (1996). Use of UV methods for measurement of protein and nucleic acid concentrations. Biotechniques, 20(6), 968-970. [CrossRef]
- 34. Burton, K. (1956). The study of the conditions and mechanisms of the diphenylamine reaction for the calorimetric estimation of deoxyribonucleic acid. The Biochemical Journal, 62(2), 615-623. [CrossRef]
- 35. Gedikoğlu, A., Öztürk, H.İ., Aytaç, E. (2023). The effect of different solvents on chemical composition, antioxidant activity, and antimicrobial potential of Turkish Cistus creticus extracts. GIDA, 48(4), 728-740. [CrossRef]
- 36. Lukas, B., Bragagna, L., Starzyk, K., Labedz, K., Stolze, K., Novak, J. (2021). Polyphenol diversity and antioxidant activity of European Cistus creticus L. (Cistaceae) compared to six further, partly sympatric Cistus species. Plants, 10(4), 615. [CrossRef]
- 37. Demetzos, C., Loukis, A., Spiliotis, V., Zoakis, N., Stratigakis, N., Katerinopoulos, H.E. (1995). Composition and antimicrobial activity of the essential oil of Cistus creticus L. Journal of Essential Oil Research, 7(4), 407-410. [CrossRef]
- 38. Demetzos, C., Stahl, B., Anastassaki, T., Gazouli, M., Tzouvelekis, L.S., Rallis, M. (1999). Chemical analysis and antimicrobial activity of the Resin Ladano, of its essential oil and of the isolated compounds. Planta Medica, 65(1), 76-78. [CrossRef]
- 39. Güvenç¸A., Yıldız, S., Özkan, A.M., Erdurak, C.S., Coşkun, M., Yılmaz, G., Okuyama, T., Okada (2005). Antimicrobiological studies on Turkish Cistus species. Pharmaceutical Biology 43(2), 178-183. [CrossRef]
- 40. Arıtuluk, Z.C. (2010). Master of Science Thesis. Flora and folk medicines of Tefenni (Burdur) District. Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
- 41. Sargin, S.A. (2021). Plants used against obesity in Turkish folk medicine: A review. Journal of Ethnopharmacology, 270, 113841. [CrossRef]