Batı anadolu'da yayılış gösteren hypecoum l. taksonları üzerinde farmasötik botanik ve biyoaktif sekonder metabolitleri yönünden araştırmalar
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Dosyalar
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Hypecoum L. cinsi Papaveraceae familyasına aittir. Bu cins ülkemizde H. procumbens L. subsp. procumbens, H. procumbens L. subsp. atropunctatum A.E.Dahl, H. pseudograndiflorum Petr., H. torulosum A.E.Dahl, H. trullatum A.E.Dahl, H. dimidiatum Delile ve H. pendulum L. olmak üzere 7 taksonla temsil edilmektedir. Bunlardan H. trullatum endemiktir. Bu familya pek çoğu ilaç etken maddesi olarak kullanılan alkaloit grupları ve sahip olduğu biyokaktivite potansiyeli açısından büyük öneme sahiptir. Ülkemizde söz konusu taksonlarla ilgili, içerdikleri alkaloit gruplarından sadece protopin üzerinde 4 adet kimyasal çalışma olup, kapsamlı çalışma bulunmadığı görülmüştür. Bu tez kapsamında Batı Anadolu'da doğal yayılış gösteren, 5 ayrı lokaliteden topladığımız ve teşhisini gerçekleştirdiğimiz 4 Hypecoum taksonu (H. procumbens subsp. procumbens, H. torulosum, H. pseudograndiflorum ve H. procumbens subsp. atropunctatum) ile çalışılmıştır. Çalışılan Hypecoum taksonları üzerinde öncelikle karşılaştırmalı morfolojik ve anatomik incelemeler gerçekleştirilip, taksonların ortak ve ayırt edici özellikleri ortaya konmuştur. Ayrıca; farklı solvan sistemleri kullanılarak çeşitli ekstraksiyon basamakları izlenmiş, daha sonra tanımlama için İTK, saflaştırma işlemleri için açık kolon kromatografisi analizleri, uçucu bileşenlerin tayini için GC-MS/MS analizleri yapılmıştır. Aynı zamanda, taksonlar üzerinde antimikrobiyal ve sitotoksik aktivite çalışmaları gerçekleştirilmiştir. Bu çalışmalarda öncelikle Hypecoum taksonlarından elde edilen ekstrelerin HeLa, MRC-5 ve A-549 hücre hatlarına karşı genel sitotoksik aktivitesi, ardından etki potansiyeli olan ekstrelerden izole edilen maddelerin PANC1 (Pankreas adenokarsinoma), MDA-MB-231 (Triple negatif invaziv meme kanseri), SH-SY5Y (Nöroblastoma) kanser hücre hatlarına karşı sitotoksik etkisi araştırılmıştır. Hypecoum procumbens subsp. procumbens'e ait etanol ekstresinin (HPPE/ON) (maserasyon) 50μg dozda HeLa hücreleri ve MRC-5 hücre hattında düşük sitotoksik aktiviteye sahip olduğu gözlenmiştir. Hypecoum procumbens subsp. procumbens'e ait metanol ekstresinin (HPPM/ON) (sıralı maserasyon), 25μg ve 50μg dozda HeLa hücreleri üzerinde doza bağımlı görece kuvvetli sitotoksik etkili olduğu saptanmıştır. Aynı ekstrenin MRC-5 hücre hattında toksik etkisi görülmemiştir. Hypecoum torulosum'a ait metanol ekstresi için (HT/M-ON) IC50 değeri 36.19 5μg/ml (Standart sapma: 0.9998) olarak belirlenmiştir. Bu ekstre HeLa hücreleri üzerinde sitotoksik aktivite göstermiş olup, MRC-5 hücre hattında herhangi bir sitotoksik etkiye rastlanmamıştır. İzole edilen maddeler için IC50 doz değerinin 10-100 ppm aralığında olduğu tespit edilmiş ve bu azalmalar istatistiksel olarak anlamlı bulunmuştur (P<0.0001). Antikanser etkilerinin doğrulanması amacıyla aynı deney düzeneğine eklenen Sisplatin'in 5μg/μl dozdaki hücre canlılığı üzerine etkileri ile karşılaştırıldığında maddelerin özellikle 100 ppm dozlarda ciddi düzeyde antikanser etkiye sahip oldukları gözlenmiştir. Söz konusu maddelerin antimikrobiyal aktivite testleri ise disk difüzyon ve mikrodilüsyon yöntemleri ile gerçekleştirilmiştir. Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Candida albicans ATCC 90029 suşları üzerinde çalışılmıştır. Bu suşlara karşı belirgin bir aktivite bulunmadığı tespit edilmiştir. H. procumbens subsp. atropunctatum taksonu hariç ülkemize ait diğer 3 taksonla ilgili morfolojik incelemelere ait tez çalışması tarafımıza aittir. Ayrıca bu çalışma ile H. torulosum, H. procumbens subsp. procumbens ve H. pseudograndiflorum taksonları anatomik açıdan tarafımızca ilk defa incelenmiş ve karakteristik yapıları ortaya konmuştur. Bu tez çalışması ile H. pseudograndiflorum ve H. torulosum bitkileri sekonder metabolitleri yönünden ilk defa incelenmiştir. Çalışılan taksonlar üzerinde antimikrobiyal aktivite deneyleri ilk kez tarafımızdan gerçekleştirilmiştir. Familyanın; Chelidonium majus, Macleaya cordata, Hylomecon maxim, Macleaya microcarpa gibi diğer bazı üyelerinden elde edilen total ekstre ve bileşikler üzerinde gerçekleştirilmiş sitotoksik aktivite çalışmaları bulunmakla birlikte; Hypecoum taksonları üzerindeki ilk sitotoksik aktivite çalışması tarafımızdan gerçekleştirilmiştir. Çalışma sonucunda elde edilen sitotoksik aktivite bulguları incelendiğinde; moleküllerin, ileri araştırmalarla umut vadeden bileşikler olarak karşımıza çıkabileceği düşünülmektedir. Hypecoum türlerinde bulunan diğer sekonder metabolitler ve biyoaktiviteleri üzerinde ileri araştırmalar yapılması gerekmektedir.
Hypecoum L. genus belongs to the Papaveraceae family. This genus is represented with 7 taxa in our country including H. procumbens L. subsp. procumbens, H. procumbens L. subsp. atropunctatum A.E.Dahl, H. pseudograndiflorum Petr., H. torulosum A.E.Dahl, H. trullatum A.E.Dahl, H. dimidiatum Delile and H. pendulum L. Among these, H. trullatum is endemic. This family is of great importance in terms of the alkaloid groups and bioactivity potential of which many of them are used as active ingredients. In Turkey, there are 4 chemical studies on the protopine, which are among the alkaloids groups, related to the taxa in question, and no comprehensive studies have been found. Within the scope of this thesis, four Hypecoum taxa (H. procumbens subsp. procumbens, H. torulosum, H. pseudograndiflorum and H. procumbens subsp. atropunctatum), which were collected from 5 different localities in Western Anatolia, were studied. Comparative morphological and anatomical investigations were carried out on the studied Hypecoum taxa and the common and distinctive features of the taxa were determined. Also; The various extraction steps were followed by using different solvent systems, then TLC for identification, open column chromatography analysis for purification procedures and GC-MS /MS analysis for determination of volatile components. Antimicrobial and cytotoxic activity studies were also performed on taxa. In these studies, the general cytotoxic activity of extracts obtained from Hypecoum taxa against HeLa, MRC-5 and A- 549 cell lines, followed by PANC1 (Pancreas adenocarcinoma), MDA-MB-231 (Triple negative invasive breast cancer) of pure substances isolated from extracts with potential for action. The cytotoxic effect of SH-SY5Y (Neuroblastoma) against cancer cell lines was investigated. Hypecoum procumbens subsp. procumbens' ethanol extract (HPPE/ON) (maceration) was observed to have low cytotoxic activity in HeLa cells and MRC-5 cell line at a dose of 50μg. Hypecoum procumbens subsp. procumbens' methanol extract (HPPM/ON) (sequential maceration) was found to have a relatively strong dose-dependent cytotoxic effect on HeLa cells at 25μg and 50μg doses. The same extract did not show toxic effects on the MRC-5 cell line. For the methanol extract of Hypecoum torulosum (HT/M-ON) the IC50 value was 36.19 5μg / ml (Standard deviation: 0.9998). This extract showed cytotoxic activity on HeLa cells and no cytotoxic effect was found on the MRC-5 cell line. The IC50 dose value for the isolated substances was found to be in the range of 10-100 ppm and these reductions were statistically significant (P <0.0001). In order to confirm the anticancer effects, cisplatin, which was added to the same experimental setup, was observed to have a significant anticancer effect especially at 100 ppm doses compared with the effects on cell viability at a dose of 5μg/μl. Antimicrobial activity tests of these substances were performed by disk diffusion and microdilution methods. Staphylococcus aureus ATCC 29213, VI Escherichia coli ATCC 25922, Candida albicans ATCC 90029 strains were studied. There was no significant activity against these strains. Except for H. procumbens subsp. atropunctatum taxa, the thesis study of the morphological studies related to 3 other taxa of our country belongs to us. In addition, H. torulosum, H. procumbens subsp. procumbens and H. pseudograndiflorum taxa have been examined for the first time in anatomical terms and their characteristic structures have been revealed. Antimicrobial activity experiments on the taxa were studied for the first time. The family; cytotoxic activity studies on total extracts and compounds obtained from some other members such as Chelidonium majus, Macleaya cordata, Macleaya microcarpa, Hylomecon maxim; The first cytotoxic activity study on Hypecoum taxa was performed by us. When the cytotoxic activity findings obtained from the study were examined; It is thought that molecules may emerge as promising compounds with further research. Further research is needed on other secondary metabolites and bioactivities found in Hypecoum species.
Hypecoum L. genus belongs to the Papaveraceae family. This genus is represented with 7 taxa in our country including H. procumbens L. subsp. procumbens, H. procumbens L. subsp. atropunctatum A.E.Dahl, H. pseudograndiflorum Petr., H. torulosum A.E.Dahl, H. trullatum A.E.Dahl, H. dimidiatum Delile and H. pendulum L. Among these, H. trullatum is endemic. This family is of great importance in terms of the alkaloid groups and bioactivity potential of which many of them are used as active ingredients. In Turkey, there are 4 chemical studies on the protopine, which are among the alkaloids groups, related to the taxa in question, and no comprehensive studies have been found. Within the scope of this thesis, four Hypecoum taxa (H. procumbens subsp. procumbens, H. torulosum, H. pseudograndiflorum and H. procumbens subsp. atropunctatum), which were collected from 5 different localities in Western Anatolia, were studied. Comparative morphological and anatomical investigations were carried out on the studied Hypecoum taxa and the common and distinctive features of the taxa were determined. Also; The various extraction steps were followed by using different solvent systems, then TLC for identification, open column chromatography analysis for purification procedures and GC-MS /MS analysis for determination of volatile components. Antimicrobial and cytotoxic activity studies were also performed on taxa. In these studies, the general cytotoxic activity of extracts obtained from Hypecoum taxa against HeLa, MRC-5 and A- 549 cell lines, followed by PANC1 (Pancreas adenocarcinoma), MDA-MB-231 (Triple negative invasive breast cancer) of pure substances isolated from extracts with potential for action. The cytotoxic effect of SH-SY5Y (Neuroblastoma) against cancer cell lines was investigated. Hypecoum procumbens subsp. procumbens' ethanol extract (HPPE/ON) (maceration) was observed to have low cytotoxic activity in HeLa cells and MRC-5 cell line at a dose of 50μg. Hypecoum procumbens subsp. procumbens' methanol extract (HPPM/ON) (sequential maceration) was found to have a relatively strong dose-dependent cytotoxic effect on HeLa cells at 25μg and 50μg doses. The same extract did not show toxic effects on the MRC-5 cell line. For the methanol extract of Hypecoum torulosum (HT/M-ON) the IC50 value was 36.19 5μg / ml (Standard deviation: 0.9998). This extract showed cytotoxic activity on HeLa cells and no cytotoxic effect was found on the MRC-5 cell line. The IC50 dose value for the isolated substances was found to be in the range of 10-100 ppm and these reductions were statistically significant (P <0.0001). In order to confirm the anticancer effects, cisplatin, which was added to the same experimental setup, was observed to have a significant anticancer effect especially at 100 ppm doses compared with the effects on cell viability at a dose of 5μg/μl. Antimicrobial activity tests of these substances were performed by disk diffusion and microdilution methods. Staphylococcus aureus ATCC 29213, VI Escherichia coli ATCC 25922, Candida albicans ATCC 90029 strains were studied. There was no significant activity against these strains. Except for H. procumbens subsp. atropunctatum taxa, the thesis study of the morphological studies related to 3 other taxa of our country belongs to us. In addition, H. torulosum, H. procumbens subsp. procumbens and H. pseudograndiflorum taxa have been examined for the first time in anatomical terms and their characteristic structures have been revealed. Antimicrobial activity experiments on the taxa were studied for the first time. The family; cytotoxic activity studies on total extracts and compounds obtained from some other members such as Chelidonium majus, Macleaya cordata, Macleaya microcarpa, Hylomecon maxim; The first cytotoxic activity study on Hypecoum taxa was performed by us. When the cytotoxic activity findings obtained from the study were examined; It is thought that molecules may emerge as promising compounds with further research. Further research is needed on other secondary metabolites and bioactivities found in Hypecoum species.
Açıklama
Anahtar Kelimeler
Papaveraceae, Hypecoum L., Alkaloit, Antimikrobiyal, Sitotoksisite, Papaveraceae, Hypecoum L., Alkaloid, Antimicrobial, Cytotoxicity
