Thymbra capitata Etanolik Ekstraktının Galleria mellonella Hemolenf Antioksidan Enzimleri Üzerine Etkisinin Değerlendirilmesi

Abstract

Acı kekik (Thymbra capitata), Akdeniz bölgesi ülkelerinde geniş yayılış göstermektedir ve medikal özelliklerinden dolayı kullanımı yaygındır. T. capitata'nın antibakteriyel, antifungal ve güçlü antioksidan özellikleri bilinmektedir. Model organizma Galleria mellonella bağışıklık araştırmaları için ve insan patojenlerinin araştırılmasında çoğunlukla tercih edilmektedir. Çalışmanın amacı, T. capitata'nın etanolik ekstraktının G. mellonella larvasında hemolenf antioksidan savunması üzerindeki etkisinin belirlenmesidir. Etanolik ekstraktan elde edilen kuru maddeden Fosfat Tampon Tuzu (PBS) ile 2 mg mL-1 ile 20 mg mL-1 arasındaki dozlarda hazırlanan çözeltiler G. mellonella larvalarına enjekte edilmiştir. Bulgularımıza göre T. capitata ekstraktının malondialdehit (MDA) düzeylerine etkisi yoktur. Ancak 10 - 20 mg mL-1 arasındaki dozların hepsinin kontrol gruplarına kıyasla süperoksit dismutaz (SOD) ve katalaz (CAT) aktivitelerini önemli ölçüde azalttığı belirlendi. Çalışmamızın sonuçlarına göre T. capitata ekstrakt yüksek dozları G. mellonella antioksidan savunması üzerinde olumsuz etkilere sahiptir.

Keywords

• Malondialdehit
• süperoksit dismutaz
• katalaz
• toplam protein

Project Number

Yok

Assessment of The Effect of Thymbra capitata Ethanolic Extract on Galleria mellonella Hemolymph Antioxidant Enzymes

Abstract

Conehead thyme (Thymbra capitata) is widely distributed in the countries of the Mediterranean region and used due to its medical properties. The antibacterial, antifungal, and strong antioxidant properties of T. capitata are known. The model organism Galleria mellonella is mostly preferred for immunological studies and for the study of human pathogens. The aim of the study was to determine the effect of the ethanolic extract of T. capitata on the antioxidant defense of the hemolymph in G. mellonella larva. Solutions prepared with Phosphate-Buffered Saline (PBS) from the dry matter obtained from ethanolic extract at doses between 2 mg mL-1 and 20 mg mL-1 were injected into G. mellonella larvae. According to our findings, T. capitata extract had no effect on malondialdehyde (MDA) levels. However, it was determined that all doses between 10 to 20 mg mL-1 significantly reduced superoxide dismutase (SOD) and catalase (CAT) activities compared to the control groups. According to the results of our study, high doses of T. capitata extract had negative effects on G. mellonella antioxidant defense.

Keywords

• Malondialdehyde
• superoxide dismutase
• catalase
• total protein.

Supporting Institution

Yok

Project Number

Yok

Thanks

The author would like to thank Begüm Çetin for her support and COMUDAM (Çanakkale Onsekiz Mart University Experimental Research Application and Research Center) staff and managers for their precious help.

References

1. Acamovic, T., & Brooker, J.D. (2005). Biochemistry of plant secondary metabolites and their effects in animals. Proceedings of the Nutrition Society, 64(3), 403-412. https://doi.org/10.1079/PNS2005449
2. Aebi, H. (1984). Catalase in vitro. Methods of Enzymology, 105, 121-126. https://doi.org/10.1016/S0076-6879(84)05016-3
3. Albano, S.M., & Miguel, M.G. (2011). Biological activities of extracts of plants grown in Portugal. Industrial Crops and Products, 33(2), 338-343. https://doi.org/10.1016/j.indcrop.2010.11.012
4. Al-Qura’n, S. (2009). Ethnopharmacological survey of wild medicinal plants in Showbak, Jordan. Journal of Ethnopharmacology, 123(1), 45-50. https://doi.org/10.1016/j.jep.2009.02.031
5. Álvarez, A., García, B.G., Jordán, M.J., Martínez-Conesa, C., & Hernández, M.D. (2012). The effect of diets supplemented with thyme essential oils and rosemary extract on the deterioration of farmed gilthead seabream (Sparus aurata) during storage on ice. Food Chemistry, 132(3), 1395-1405. https://doi.org/10.1016/j.foodchem.2011.11.127
6. Barbosa, P., Lima, A.S., Vieira, P., Dias, L.S., Tinoco, M. T., Barroso, J. G., ... & Mota, M. (2010). Nematicidal activity of essential oils and volatiles derived from Portuguese aromatic flora against the pinewood nematode, Bursaphelenchus xylophilus. Journal of Nematology, 42(1), 8.
7. Blanco-Salas, J., Téllez, T.R., Alonso, M.J.P., Pardo, F.M.V., de los Ángeles Cases Capdevila, M., & Gervasini Rodríguez, C. (2010). Chemical composition and antioxidant activity of the essential oil of Thymbra capitata (L.) Cav. in Spain. Acta Botanica Gallica, 157 (1), 55-63. https://doi.org/10.1080/12538078.2010.10516189
8. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72(1-2), 248-254. https://doi.org/10.1016/0003-2697(76)90527-3

9. Buege J.A., & Aust S.D. (1978). Microsomal Lipid Peroxidation, Methods in Enzymology, 52: 302-310. https://doi.org/10.1016/S0076-6879(78)52032-6
10. Büyükgüzel, E., Büyükgüzel, K., Snela, M., Erdem, M., Radtke, K., Ziemnicki, K., & Adamski, Z. (2013). Effect of boric acid on antioxidant enzyme activity, lipid peroxidation, and ultrastructure of midgut and fat body of Galleria mellonella. Cell biology and toxicology, 29(2), 117-129. https://doi.org/10.1007/s10565-013-9240-7
11. Büyükgüzel, E., Hyršl, P., & Büyükgüzel, K. (2010). Eicosanoids mediate hemolymph oxidative and antioxidative response in larvae of Galleria mellonella L. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 156(2), 176-183. https://doi.org/10.1016/j.cbpa.2010.01.020
12. Büyükgüzel E., & Kayaoğlu S. (2014). The effect of niclosamide on some biological and physiological aspects of Galleria mellonella L. (Lepidoptera: Pyralidae). Türkiye Entomoloji Dergisi, 38(1), 83-100. https://doi.org/10.16970/ted.87976
13. Cook S.M., & McArthur J.D. (2013). Developing Galleria mellonella as a model host for human pathogens. Virulence, 4, 350-353. https://doi.org/10.4161/viru.25240
14. Dere, B., Altuntaş, H., & Nurullahoğlu, Z.U. (2015). Insecticidal and oxidative effects of azadirachtin on the model organism Galleria mellonella L. (Lepidoptera: Pyralidae). Archives of insect biochemistry and physiology, 89(3), 138-152. https://doi.org/10.1002/arch.21231
15. Du E, Wang W, Gan L, Li Z, Guo S., & Guo Y. (2016). Effects of thymol and carvacrol supplementation on intestinal integrity and immune responses of broiler chickens challenged with Clostridium perfringens. Journal of animal science and biotechnology. 7: Article number: 19. https://doi.org/10.1186/s40104-016-0079-7
16. Dubovskii, I.M., Grizanova, E.V., Chertkova, E.A., Slepneva, I.A., Komarov, D.A., Vorontsova, Y.L., & Glupov, V.V. (2010). Generation of reactive oxygen species and activity of antioxidants in hemolymph of the moth larvae Galleria mellonella (L.) (Lepidoptera: Piralidae) at development of the process of encapsulation. Journal of Evolutionary Biochemistry and Physiology, 46(1), 35-43. https://doi.org/10.1134/S0022093010010044
17. Dubovskiy, I.M., Martemyanov, V.V., Vorontsova, Y.L., Rantala, M.J., Gryzanova, E.V., & Glupov, V.V. (2008). Effect of bacterial infection on antioxidant activity and lipid peroxidation in the midgut of Galleria mellonella L. larvae (Lepidoptera, Pyralidae). Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 148(1), 1-5. https://doi.org/10.1016/j.cbpc.2008.02.003
18. Eguchi D., & Iwabuchi K. (2006). A new cell line from the wax moth Galleria mellonella Linne (Lepidoptera: Pyralididae). In Vitro Cellular & Developmental Biology-Animal, 42, 1-3. https://doi.org/10.1007/s11626-006-0002-1
19. Ercan, F.S., Baş, H., & Azarkan, S Y. (2022). In silico detection of Cucurbitacin-E on antioxidant enzymes of model organism Galleria mellonella L.(Lepidoptera: Pyralidae) and variation of antioxidant enzyme activities and lipid peroxidation in treated larvae. Beni-Suef University Journal of Basic and Applied Sciences, 11(1), 1-17. https://doi.org/10.1186/s43088-022-00310-3
20. Faleiro, L., Miguel, G., Gomes, S., Costa, L., Venâncio, F., Teixeira, A., Figueiredo, A.C., Barroso J.G., & Pedro, L. G. (2005). Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L.(Cav.) and Origanum vulgare L. Journal of Agricultural and Food Chemistry, 53(21), 8162-8168. https://doi.org/10.1021/jf0510079
21. Flöhe L., & Ötting F. (1984). Superoxide Dismutase Assays. Methods of Enzymology, 105, 93-104. https://doi.org/10.1016/S0076-6879(84)05013-8
22. Gagliano Candela, R., Maggi, F., Lazzara, G., Rosselli, S., & Bruno, M. (2019). The Essential Oil of Thymbra capitata and its Application as A Biocide on Stone and Derived Surfaces. Plants, 8(9), 300. https://doi.org/10.3390/plants8090300
23. Hashemipour H, Kermanshahi H, Golian A, & Veldkamp T. (2013). Effect of thymol and carvacrol feed supplementation on performance, antioxidant enzyme activities, fatty acid composition, digestive enzyme activities, and immune response in broiler chickens. Poultry science, 92, 2059-2069. https://doi.org/10.3382/ps.2012-02685
24. Hortigón-Vinagre, M.P., Blanco, J., Ruiz, T., & Henao, F. (2014). Thymbra capitata essential oil prevents cell death induced by 4-hydroxy-2-nonenal in neonatal rat cardiac myocytes. Planta medica, 80(15), 1284-1290. https://doi.org/10.1055/s-0034-1383051
25. Hyršl, P., Büyükgüzel, E., & Büyükgüzel, K. (2007). The effects of boric acid-induced oxidative stress on antioxidant enzymes and survivorship in Galleria mellonella. Archives of Insect Biochemistry and Physiology: Published in Collaboration with the Entomological Society of America, 66(1), 23-31. https://doi.org/10.1002/arch.20194
26. Kaya, S., Akkuş, G., Türkdoğan, S., & Gündüz, B. (2021). Influence of Helichrysum arenarium on hemocyte-mediated immune responses and phenoloxidase enzyme activity of model organism Galleria mellonella (L.). International Journal of Tropical Insect Science, 41(4), 2521-2528. https://doi.org/10.1007/s42690-021-00432-8
27. Kaya, S., & Demir, N. (2020). Zeytin (Olea europaea) Yaprağı Ekstraktının Model Organizma Galleria mellonella Hemosit Aracılı Bağışıklık Tepkileri Üzerine Etkileri. Türk Tarım ve Doğa Bilimleri Dergisi, 7(3), 646-653. https://doi.org/10.30910/turkjans.691138
28. Khan, I., Bahuguna, A., Kumar, P., Bajpai, V.K., & Kang, S.C. (2018). In vitro and in vivo antitumor potential of carvacrol nanoemulsion against human lung adenocarcinoma A549 cells via mitochondrial mediated apoptosis. Scientific Reports, 8(1), 144. https://doi.org/10.1038/s41598-017-18644-9
29. Leonelli, S., & Ankeny, R.A. (2013). What makes a model organism? Endeavour, 37(4), 209-212. https://doi.org/10.1016/j.endeavour.2013.06.001
30. Lozinskaya, Y.L., Slepneva, I.A., Khramtsov, V.V., & Glupov, V.V. (2004). Changes of the antioxidant status and system of generation of free radicals in hemolymph of Galleria mellonella larvae at microsporidiosis. Journal of Evolutionary Biochemistry and Physiology, 40(2):119-125. https://doi.org/10.1023/B:JOEY.0000033802.97996.65
31. Machado, M., Dinis, A. M., Salgueiro, L., Cavaleiro, C., Custódio, J. B., & do Céu Sousa, M. (2010). Anti-Giardia activity of phenolic-rich essential oils: effects of Thymbra capitata, Origanum virens, Thymus zygis subsp. sylvestris, and Lippia graveolens on trophozoites growth, viability, adherence, and ultrastructure. Parasitology Research, 106(5), 1205-1215. https://doi.org/10.1007/s00436-010-1800-7
32. Marchese, A., Arciola, C.R., Coppo, E., Barbieri R., Barreca D., & Chebaibi S. (2018). The natural plant compound carvacrol as an antimicrobial and anti-biofilm agent: mechanisms, synergies and bio-inspired anti-infective materials. Biofouling. 34, 630-56. https://doi.org/10.1080/08927014.2018.1480756
33. Marinelli, L., Di Stefano, A., & Cacciatore, I. (2018). Carvacrol and its derivatives as antibacterial agents. Phytochemistry Reviews, 17(4), 903-921. https://doi.org/10.1007/s11101-018-9569-x
34. Mbarga, M. J. A., Podoprigora, I. V., & Anyutoulou, K. L. D. (2021). Galleria mellonella (greater wax moth) as an eco-friendly in vivo approach for the assessment of the acute toxicity of medicinal plants: Application to some plants from Cameroon. Open Veterinary Journal, 11(4), 651-661. https://doi.org/10.5455/OVJ.2021.v11.i4.15
35. Mukherjee, K., Altincicek, B., Hain, T., Domann, E., Vilcinskas, A., & Chakraborty, T. (2010). Galleria mellonella as a Model System for Studying Listeria Pathogenesis. Applied And Environmental Microbiology, 76 (1), 310–317. https://doi.org/10.1128/AEM.01301-09
36. Palmeira-de-Oliveira, A., Gaspar, C., Palmeira-de-Oliveira, R., Silva-Dias, A., Salgueiro, L., Cavaleiro, C. & Rodrigues, A.G. (2012). The anti-Candida activity of Thymbra capitata essential oil: effect upon pre-formed biofilm. Journal of Ethnopharmacology, 140(2), 379-383. https://doi.org/10.1016/j.jep.2012.01.029
37. Saija, A., Speciale, A., Trombetta, D., Leto, C., Tuttolomondo, T., La Bella, S., & Napoli, E. (2016). Phytochemical, Ecological and Antioxidant Evaluation of Wild Sicilian Thyme: Thymbra capitata (L.) Cav. Chemistry & Biodiversity, 13(12), 1641-1655. https://doi.org/10.1002/cbdv.201600072
38. Sak O., Uçkan F., & Ergin E., (2006). Effects of cypermethrin on total body weight, glycogen, protein, and lipid contents of Pimpla turionellae (L.) (Hymenoptera: Ichneumonidae), Belgian Journal of Zoology, 136, 53-58.
39. Silveira S.R.C., Andrade L.N., & Sousa D.P. (2013). A Review on Anti-Inflammatory Activity of Monoterpenes. Molecules. 18, 1227-1254. https://doi.org/10.3390/molecules18011227
40. Vila, R. (2002). Flavonoids and further polyphenols in the genus Thymus (pp. 144-176). in ed: Stahl- Biskup, E., & Sáez, F. Thyme: the genus Thymus. CRC Press. Taylor & Francis: New York.
41. Wu, G., & Yi, Y. (2015). Effects of dietary heavy metals on the immune and antioxidant systems of Galleria mellonella larvae. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 167, 131-139. https://doi.org/10.1016/j.cbpc.2014.10.004
42. Yu, Y.M., Chao, T.Y., Chang, W.C., Chang, M.J., & Lee, M.F. (2016). Thymol reduces oxidative stress, aortic intimal thickening, and inflammation-related gene expression in hyperlipidemic rabbits. Journal of Food and Drug Analysis, 24(3), 556-563. https://doi.org/10.1016/j.jfda.2016.02.004
43. Zorlu, T., Nurullahoğlu, Z.U., & Altuntaş, H. (2018). Influence of dietary titanium dioxide nanoparticles on the biology and antioxidant system of model insect, Galleria mellonella (L.) (Lepidoptera: Pyralidae). Journal of the Entomological Research Society, 20(3), 89-103.