Capsaicin’in yağ doku kökenli mezenkimal kök hücreler üzerine etkisi
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Date
2021-12-01
Authors
İncebıyık, Ece
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Canlı organ ve dokularda, kaynak hücre olarak uzun süre canlığını sürdüren, kendini yenileyebilen, farklılaşarak diğer doku hücrelerine dönüşebilen, henüz farklılaşmamış hücreler kök hücre olarak tanımlanır. Erişkin memeli canlıdaki mezenkimal kök hücreler (MKH) kolaylıkla ve yüksek oranda bulunan multipotent kök hücrelerdir. Kırmızı acı biberin etken maddesi olan Capsaicin (8-methyl-Nvanillyl-trans-6-nonenamide) (CAP), alkoloid yapıya sahiptir. Günümüz koşullarında farklı kullanım alanlarına sahip kırmızı acı biber, başlangıçta gıda katkı maddesi olarak kullanılmıştır. CAP’ın 19. yüzyıldan itibaren analjezik etkisinden yararlanılmaya başlanmıştır. Günümüzde araştırma ve ilaç sektöründe kullanılmaktadır. Bu tez çalışmasında; sıçan yağ dokusundan elde edilen mezenkimal kök hücrelerine farklı doz ve sürelerde uygulanan Capsaicin’in, Ki-67 antikoru ile proliferasyonuna, kök hücre yüzey işaretleyicileri (cluster differentiation CD), CD90, CD105, CD45 ve CD11b ile fenotipik karakterizasyonuna ve MTT testi ile hücre sağ kalımına olası etkisinin değerlendirilmesi amaçlandı. Mezenkimal kök hücre (MKH) izolasyonu sonucu elde edilen pasaj 3 hücrelerinin CD90 ve CD105 antikorları ile pozitif; CD11b ve CD45 antikorları ile negatif immunreaksiyonu tanımlandı. Aynı zamanda pasaj 3 hücrelerinin osteoblast, kondroblast ve adiposit hücrelerine farklılaştırmaları gerçekleştirildi. Daha sonra pasaj 3 MKH hücrelerine 0, 5, 10, 25, 50, 100, 150 µM CAP dozları 24 ve 48 saat süre ile uygulandı. Bu hücrelerin MKH özelliklerine CAP’ın etkisi; Ki-67, CD90, CD105, CD11b ve CD45 antikorlarının immunreaksiyonları ile değerlendirildi. CAP uygulamasının hücrelerdeki sağ kalım düzeyi ise MTT testi ile incelendi. Sonuç olarak; yağ doku kökenli pasaj 3 mezenkimal kök hücrelerinde; 24 saat süre ile 5, 10 ve 25 µM CAP dozlarının mezenkimal kök hücre fenotipini değiştirmeden kök hücre proliferasyonunu ve kök hücre sağ kalımını arttırdığı saptandı. Ancak 24 ve 48 saatlik 50 µM ve üzeri uygulanan CAP dozlarının hücresel farklılaşmayı indüklediği, hücre canlılığı ve proliferasyonu azalttığı belirlendi.
Stem cells are defined as the cells that have the ability to renew themselves, differentiate into other tissue cells and survive for a long time as source cells in living organ and tissues. Mesenchymal stem cells (MSCs) in adult mammals are multipotent stem cells that are readily and highly prevalent in mammals. Capsaicin (8-methyl-N-vanillyl-trans-6-nonenamide), the active ingredient of red hot chili pepper, (CAP) has an alkaloid structure. Red hot chili pepper, which has different uses nowadays, was initially used as a food additive. CAP has been used for its analgesic effect since the 19th century. It is currently being utilized in research and pharmaceutical industry. In this thesis study; the effect of CAP on cell proliferation using Ki-67 antibody, phenotypic characterization with stem cell surface markers, cluster differentiation, CD90, CD105, CD45 and CD11b, and on cell survival using MTT test in mesenchymal stem cells obtained from rat adipose tissue at different doses and times. Positive immunoreaction of CD90 and CD105 and negative immunoreaction of CD11b and CD45 markers were determined in passage 3 cells that were obtained by mesenchymal stem cell (MSC) isolation. At the same time, differentiations of osteoblast, chondroblast and adipocyte in passage 3 cells were determined. Then, 0, 5, 10, 25, 50, 100, 150 µM CAP doses were applied to passage 3 MSC cells for 24 and 48 hours. The effect of CAP on MSC properties of these cells was evaluated by immunoreactions of Ki-67, CD90, CD105, CD11b and CD45 antibodies. Cellular survival upon CAP treatment was examined by MTT test. As a result; in adipose tissue-derived passage 3 mesenchymal stem cells, it was determined that CAP at 5, 10 and 25 µM CAP doses for 24 hours increased stem cell proliferation and stem cell survival without affecting the mesenchymal stem cell phenotype. However, it was determined that CAP at doses of 50 µM and above applied for 24 and 48 hours induced cellular differentiation while decreasing cell viability and proliferation.
Stem cells are defined as the cells that have the ability to renew themselves, differentiate into other tissue cells and survive for a long time as source cells in living organ and tissues. Mesenchymal stem cells (MSCs) in adult mammals are multipotent stem cells that are readily and highly prevalent in mammals. Capsaicin (8-methyl-N-vanillyl-trans-6-nonenamide), the active ingredient of red hot chili pepper, (CAP) has an alkaloid structure. Red hot chili pepper, which has different uses nowadays, was initially used as a food additive. CAP has been used for its analgesic effect since the 19th century. It is currently being utilized in research and pharmaceutical industry. In this thesis study; the effect of CAP on cell proliferation using Ki-67 antibody, phenotypic characterization with stem cell surface markers, cluster differentiation, CD90, CD105, CD45 and CD11b, and on cell survival using MTT test in mesenchymal stem cells obtained from rat adipose tissue at different doses and times. Positive immunoreaction of CD90 and CD105 and negative immunoreaction of CD11b and CD45 markers were determined in passage 3 cells that were obtained by mesenchymal stem cell (MSC) isolation. At the same time, differentiations of osteoblast, chondroblast and adipocyte in passage 3 cells were determined. Then, 0, 5, 10, 25, 50, 100, 150 µM CAP doses were applied to passage 3 MSC cells for 24 and 48 hours. The effect of CAP on MSC properties of these cells was evaluated by immunoreactions of Ki-67, CD90, CD105, CD11b and CD45 antibodies. Cellular survival upon CAP treatment was examined by MTT test. As a result; in adipose tissue-derived passage 3 mesenchymal stem cells, it was determined that CAP at 5, 10 and 25 µM CAP doses for 24 hours increased stem cell proliferation and stem cell survival without affecting the mesenchymal stem cell phenotype. However, it was determined that CAP at doses of 50 µM and above applied for 24 and 48 hours induced cellular differentiation while decreasing cell viability and proliferation.
Description
Keywords
Kapsaisin, Kapsaisinoidler, Kök hücre taransplantasyonu, Kök hücreler, Capsaicin, Capsaicinoids, Stem cell transplantation, Stem cells
Citation
İncebıyık, E. (2021). Capsaicin’in yağ doku kökenli mezenkimal kök hücreler üzerine etkisi. Yayınlanmamış doktora tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.