Beta-glukan ve Cephalaria saponin kombinasyonlarının immünomodülatör etkinliğinin araştırılması
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Tarih
2021
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Yayıncı
Ege Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Doğal immün modülatörler, bağışıklık sistemini etkileyen bozuklukları kontrol etmek ve onarmak için anahtar olmaktadır. Saponinlerin, bağışıklık uyarıcı etkilerine aracılık edebilecek interlökinler ve interferonlar gibi sitokinlerin üretimini indüklediği yapılan çalışmalarda bildirilmiştir. β-glukanların makrofajlar, dendritik, nötrofiller, B hücreleri, T hücreleri ve doğal öldürücü hücreleri aktive ettiği bilinmektedir. Yapılan tez çalışmasında Cephalaria spp. izole edilen Scoposide A, Scoposide B, Aristatoside B, Elmalienoside A, Balansoide B saponin moleküllerinin, β-glukan ve β-glukan saponin kombinasyonlarının THP-1 makrofaj hücresi üzerindeki immünomodülatör etkinliğinin araştırılması amaçlanmıştır. THP-1 monosit hücreleri PMA ile farklılaştırılmış ve saponin molekülleri, β-glukan ve β-glukan saponin kombinasyonları ile 24 saatlik inkübasyonu ardından makrofaj hücrelerinde FITC-CD11b, FITC-CD163 ve PE-CD80 antikorları ile boyanarak makrofaj polarizasyonuna etkileri flow sitometri cihazında belirlenmiştir. Çalışma sonucunda β-glukan (3µg/ml) CD80 ekspresyon seviyesi %34.3±1.77 olarak elde edilmiştir. Scoposide B (3µg/ml) ve β-glukan (3µg/ml) kombinasyonu CD80 ekspresyonu %25.4±0.26 (p<0.0001) olarak elde edilmiştir. Scoposide A saponin molekülü β-glukan ile kombinasyonunda CD 163 ekspresyonu %8.8± 0.29 (p>0.5), CD80 ekspresyonu ise %8.1±0.21 (p<0.0001) olarak elde edilmiştir. β-glukan-Elmalienoside A kombinasyonun CD 163 ve CD80 ekspresyon yüzde değerlerine göre Th2 yantını desteklediği görülmüştür. Çalışma sonuçlarında, β-glukan ve saponin β-glukan kombinasyonunun makrofaj orijinli THP-1 hücrelerinde hem pro-inflamatuar hem de anti-inflamatuar etkiler görülmüştür. Çalışma sonuçları tümör boyutunun küçülmesi, kanser çalışmaları, temel immün tepkisi seviyesini arttırarak immünoterapötik ajan olarak immün yanıtı bozulmuş bireyler için β-glukan ve saponin kombinasyonları potansiyel olabilecektir.
Innate immune modulators are key to controlling and repairing damage that affect the immune system. Studies have reported that saponins induce the production of cytokines such as interleukins and interferons that may mediate their immunostimulating effects. β-glucans are known to activate macrophages, dendritic, neutrophils, B cells, T cells and natural killer cells. In this thesis, it was aimed to investigate the immunomodulatory activity of Scoposide A, Scoposide B, Aristatoside B, Elmalienoside A, Balanoide B saponin molecules isolated from Cephalaria spp., β-glucan and β-glucan saponin combinations on THP-1 macrophage cell. THP-1 monocyte cells were differentiated to THP-1 macrophage with PMA incubation.Then, Scoposide A, Scoposide B, Aristatoside B, Elmalienoside A, Balanoide B, β-glucan and β-glucan saponin combinations are added on macrophage. After 24 hours incubation, by staining macrophage cells with FITC-CD11b, FITC-CD163 and PE-CD80 antibodies and effect of macrophage polarization in flow cytometer has been determined. As a result, β-glucan (3μg/ml) CD80 expression level was obtained as 34.3±1.77%. The combination of Scoposide B (3μg/ml) and β-glucan (3μg/ml) resulted in CD80 expression level of 25.4±0.26% (p<0.0001). In the combination of Scoposide A saponin molecule with β-glucan, CD163 expression was 8.8±0.29% (p>0.5), and CD80 expression was 8.1±0.21% (p<0.0001). It was observed that the combination of β-glucan-Elmalienoside A supported the Th2 response according to CD163 and CD80 expression percentage levels. The conclusion, it was observed that the combination of β-glucan and saponin β-glucan both promoted pro-inflammatory and anti-inflammation in THP-1 cells of macrophage origin. The results of the study may be potential that β-glucan and saponin combinations for individuals with impaired immune response as immunotherapeutic agents by reducing tumor size, cancer studies, increasing the level of basic immune response.
Innate immune modulators are key to controlling and repairing damage that affect the immune system. Studies have reported that saponins induce the production of cytokines such as interleukins and interferons that may mediate their immunostimulating effects. β-glucans are known to activate macrophages, dendritic, neutrophils, B cells, T cells and natural killer cells. In this thesis, it was aimed to investigate the immunomodulatory activity of Scoposide A, Scoposide B, Aristatoside B, Elmalienoside A, Balanoide B saponin molecules isolated from Cephalaria spp., β-glucan and β-glucan saponin combinations on THP-1 macrophage cell. THP-1 monocyte cells were differentiated to THP-1 macrophage with PMA incubation.Then, Scoposide A, Scoposide B, Aristatoside B, Elmalienoside A, Balanoide B, β-glucan and β-glucan saponin combinations are added on macrophage. After 24 hours incubation, by staining macrophage cells with FITC-CD11b, FITC-CD163 and PE-CD80 antibodies and effect of macrophage polarization in flow cytometer has been determined. As a result, β-glucan (3μg/ml) CD80 expression level was obtained as 34.3±1.77%. The combination of Scoposide B (3μg/ml) and β-glucan (3μg/ml) resulted in CD80 expression level of 25.4±0.26% (p<0.0001). In the combination of Scoposide A saponin molecule with β-glucan, CD163 expression was 8.8±0.29% (p>0.5), and CD80 expression was 8.1±0.21% (p<0.0001). It was observed that the combination of β-glucan-Elmalienoside A supported the Th2 response according to CD163 and CD80 expression percentage levels. The conclusion, it was observed that the combination of β-glucan and saponin β-glucan both promoted pro-inflammatory and anti-inflammation in THP-1 cells of macrophage origin. The results of the study may be potential that β-glucan and saponin combinations for individuals with impaired immune response as immunotherapeutic agents by reducing tumor size, cancer studies, increasing the level of basic immune response.
Açıklama
Anahtar Kelimeler
β-Glukan, Saponin, Cephalaria spp., İmmünomodülatör, β-Glucan, Saponin, Cephalaria spp., Immunomodulator
