Arı ekmeğinin biyokimyasal, duyusal karakterizasyonu ve biyolojik aktivitesinin in vitro belirlenmesi
Küçük Resim Yok
Tarih
2025
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Bursa Teknik Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Geçmişten günümüze yapılan araştırmalar, doğal arı ürünlerinin birçok hastalığa karşı koruyucu olarak veya tedavi amaçlı kullanılabileceğini göstermektedir. Bu anlamda geleneksel, tamamlayıcı ve fonksiyonel tıp uygulamaları arasında yer alan apiterapi, arı ürünlerinin (bal, balmumu, propolis, arı poleni, arı sütü, arı ekmeği, arı zehri, apilarnil ve arı kovanı havası) kullanıldığı doğal bir tedavi yöntemidir. Apiterapide kullanılan ve arı ürünleri arasında nispeten az bilinen arı ekmeği (perga); arılar tarafından toplanan çiçek nektarı ve polenin arının kendi vücut salgıları, bal ve balmumu ile karıştırılıp petek gözlerine depolanması burada bakteri ve mayaların yardımıyla anaerobik şartlarda laktik asit fermantasyonu sonucu oluşan bir üründür. Bu çalışmada son yıllarda ilgi çeken arı ekmeğinin hem fizikokimyasal, biyokimyasal ve duyusal özelliklerinin belirlenmesi; hem de biyolojik aktivitesine yönelik olarak MTT ve NKA analizleri ile sitotoksik, COMET ve MN testi ile genotoksik ve antigenotoksik, total 5mC tayini ile epigenetik etkilerinin araştırılması amaçlanmıştır. Aroma maddelerinin ve yüzde oranları GC-MS cihazında SPME tekniği kullanılarak belirlenmiştir. Arı ekmeğinin toplam protein içeriği %19,06; nem içeriği %17,89; kül içeriği %2,53; yağ içeriği %9,16 olarak bulunmuştur. Arı ekmeğinin toplam fenolik içeriği 9,91 mg GAE/g, toplam flavonoid içeriği 0,32 mg QE/g olarak bulunmuştur. Arı ekmeğinin antioksidan kapasite analizlerine göre ABTS.+ kapasite değeri 0,55 mM Trolox/mL, CUPRAC değeri 12,97 mg Trolox/g olarak belirlenmiştir. Yapılan çalışmalarda elde edilen sonuçlar, geçmişte yapılan diğer çalışmaların sonuçlarıyla karşılaştırıldığında küçük farklılıklar olduğu görülse de sonuçların birbirini destekler nitelikte olduğu saptanmıştır. Arı ekmeğinin fizikokimyasal, antioksidan kapasite ve duyusal karakterizasyon özellikleri ürünün toplandığı bölge, iklim, bitki örtüsü, toplanma ve depolanma şekli, arının türü, arı kolonisinin durumu gibi birçok parametreye bağlı olarak değişiklik göstermektedir. Biyolojik aktivitesinin belirlenmesi amacıyla BEAS-2B ve A549 hücreleri üzerinde yapılan MTT ve NKA sitotoksisite test sonuçlarına göre arı ekmeğinin uygulama konsantrasyonlarının (6,25-800 µg/mL) sitotoksik etki göstermediği belirlenmiştir. Comet testi ile değerlendirilen genotoksisite sonuçlarına göre, 100, 200, 400, 800 µg/mL konsantrasyonda uygulanan arı ekmeğinin BEAS-2B ve A549 hücrelerinde muamelesiz negatif kontrol grubuna kıyasla genotoksik etki oluşturmadığı, diğer yandan H2O2 ile DNA hasarına uğratılan hücrelerde antigenotoksik etkisi değerlendirildiğinde tek başına H2O2 uygulanan pozitif kontrole göre hasarı azalttığı belirlenmiştir. MN testi ile değerlendirilen genotoksisite sonuçlarına göre, 100, 200, 400, 800 µg/mL konsantrasyonda uygulanan arı ekmeğinin insan lenfosit hücrelerinde muamelesiz negatif kontrol grubuna kıyasla istatistiksel olarak anlamlı bir artışa neden olmadığı yani genotoksik etki oluşturmadığı, diğer yandan H2O2 ile DNA hasarına uğratılan hücrelerde antigenotoksik etkisi değerlendirildiğinde tek başına H2O2 uygulanan pozitif kontrole göre hasarı azalttığı belirlenmiştir. Arı ekmeğinin BEAS-2B hücrelerinde 24 saat süre ile uygulanmasının ardından tüm uygulama gruplarında kontrol grubuna kıyasla total genomik DNA metilasyonunda azalma olduğu belirlenmiştir. Elde edilen sonuçlar, arı ekmeğinin güçlü antioksidan aktivitesine bağlı hücre ve DNA hasarını azaltıcı aktivite gösterdiğini ortaya koymuştur.
Past and present research demonstrates that natural bee products can be used as preventative or therapeutic against many diseases. Apitherapy, a traditional, complementary, and functional medicine practice, is a natural treatment method that utilizes bee products (honey, beeswax, propolis, bee pollen, royal jelly, bee bread, bee venom, apilarnil, and beehive air). Bee bread (perga), a relatively lesser-known bee product used in apitherapy, is a product of the mixture of flower nectar and pollen collected by bees with their own bodily secretions, honey, and beeswax, which is then stored in honeycomb cells and subsequently formed through lactic acid fermentation under anaerobic conditions with the help of bacteria and yeast. In this study, it was aimed to determine the physicochemical, biochemical and sensory properties of bee bread, which has attracted attention in recent years, and to investigate its biological activity by cytotoxicity with MTT and NRU analyses, genotoxicity and antigenotoxicity with COMET and MN tests, and epigenetic effects with total 5mC determination. Identification of aroma substances and their percentage ratios were carried out using the SPME technique on the GC-MS device. The total protein content of bee bread was found as 19.06%; moisture content was found as 17.89%; ash content was found as 2.53%; fat content was found as 9.16%. Total phenolic content of bee bread was found as 9.91 mg GAE/g, total flavonoid content was found as 0.32 mg QE/g. According to antioxidant capacity analyses of bee bread, ABTS.+ capacity value was determined as 0.55 mM Trolox/mL, CUPRAC value was determined as 12.97 mg Trolox/g. When the results obtained in the study were compared with the results of previous studies, it was determined that although there were minor differences, the results supported each other. The physicochemical, antioxidant capacity and sensory characterization properties of bee bread vary depending on many parameters such as the region where the product is collected, climate, vegetation, collection and storage method, bee species, and the condition of the bee colony. In order to evaluate its biological activity, it was determined that the exposure concentrations of bee bread (6.25-800 µg/mL) did not show cytotoxic effect according to the MTT and NRU cytotoxicity tests performed on BEAS-2B and A549 cells. According to the genotoxicity results evaluated with the comet test, bee bread applied at concentrations of 100, 200, 400, 800 µg/mL did not show a genotoxic effect on BEAS-2B and A549 cells compared to the untreated negative control group, on the other hand, when its antigenotoxic effect was evaluated in cells subjected to DNA damage with H2O2, it was determined that it reduced the damage compared to the positive control. According to the genotoxicity results evaluated by the MN test, bee bread applied at concentrations of 100, 200, 400, and 800 µg/mL did not cause a statistically significant increase in human lymphocyte cells compared to the untreated negative control group and did not cause a genotoxic effect. On the other hand, when its antigenotoxic effect was evaluated in cells subjected to DNA damage with H2O2, it reduced the damage compared to the positive control treated with H2O2 alone. Following 24-hour application of bee bread to BEAS-2B cells, a decrease in total genomic DNA methylation was determined in all treatment groups compared to the control group. The results obtained revealed that bee bread exhibits cell and DNA damage-reducing activity due to its strong antioxidant activity.
Past and present research demonstrates that natural bee products can be used as preventative or therapeutic against many diseases. Apitherapy, a traditional, complementary, and functional medicine practice, is a natural treatment method that utilizes bee products (honey, beeswax, propolis, bee pollen, royal jelly, bee bread, bee venom, apilarnil, and beehive air). Bee bread (perga), a relatively lesser-known bee product used in apitherapy, is a product of the mixture of flower nectar and pollen collected by bees with their own bodily secretions, honey, and beeswax, which is then stored in honeycomb cells and subsequently formed through lactic acid fermentation under anaerobic conditions with the help of bacteria and yeast. In this study, it was aimed to determine the physicochemical, biochemical and sensory properties of bee bread, which has attracted attention in recent years, and to investigate its biological activity by cytotoxicity with MTT and NRU analyses, genotoxicity and antigenotoxicity with COMET and MN tests, and epigenetic effects with total 5mC determination. Identification of aroma substances and their percentage ratios were carried out using the SPME technique on the GC-MS device. The total protein content of bee bread was found as 19.06%; moisture content was found as 17.89%; ash content was found as 2.53%; fat content was found as 9.16%. Total phenolic content of bee bread was found as 9.91 mg GAE/g, total flavonoid content was found as 0.32 mg QE/g. According to antioxidant capacity analyses of bee bread, ABTS.+ capacity value was determined as 0.55 mM Trolox/mL, CUPRAC value was determined as 12.97 mg Trolox/g. When the results obtained in the study were compared with the results of previous studies, it was determined that although there were minor differences, the results supported each other. The physicochemical, antioxidant capacity and sensory characterization properties of bee bread vary depending on many parameters such as the region where the product is collected, climate, vegetation, collection and storage method, bee species, and the condition of the bee colony. In order to evaluate its biological activity, it was determined that the exposure concentrations of bee bread (6.25-800 µg/mL) did not show cytotoxic effect according to the MTT and NRU cytotoxicity tests performed on BEAS-2B and A549 cells. According to the genotoxicity results evaluated with the comet test, bee bread applied at concentrations of 100, 200, 400, 800 µg/mL did not show a genotoxic effect on BEAS-2B and A549 cells compared to the untreated negative control group, on the other hand, when its antigenotoxic effect was evaluated in cells subjected to DNA damage with H2O2, it was determined that it reduced the damage compared to the positive control. According to the genotoxicity results evaluated by the MN test, bee bread applied at concentrations of 100, 200, 400, and 800 µg/mL did not cause a statistically significant increase in human lymphocyte cells compared to the untreated negative control group and did not cause a genotoxic effect. On the other hand, when its antigenotoxic effect was evaluated in cells subjected to DNA damage with H2O2, it reduced the damage compared to the positive control treated with H2O2 alone. Following 24-hour application of bee bread to BEAS-2B cells, a decrease in total genomic DNA methylation was determined in all treatment groups compared to the control group. The results obtained revealed that bee bread exhibits cell and DNA damage-reducing activity due to its strong antioxidant activity.
Açıklama
Anahtar Kelimeler
Biyoteknoloji, Biotechnology
