Bal arılarında (Apis mellifera) beslenme farklılıklarının sirtuin enzimleri ve nörodejeneratif ve nörotransmitter salınımını düzenleyen genler üzerinde etkilerinin araştırılması
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Date
2024
Authors
Çelik, Sevda
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Publisher
Bursa Uludağ Üniversitesi
Abstract
Nörodejeneratif hastalıkların kesin tedavisi henüz bulunmamaktadır. Birçok nörodejeneratif bozuklukların araştırılmasında model organizma olarak bal arıları kullanılmaktadır. Histon deasetilaz olan sirtuinlerin nörodejeneratif hastalıklara katkı sunduğu bilinmektedir fakat hangi gen ve yolaklar üzerinden olduğu bilinmediğinden bu tez çalışmasında bal arılarında, SIRT1, SIRT6 ve SIRT7 aktivitesi ve bununla ilişkili nörotransmitter salınımını kontrol eden BRP geni, nörodejeneratif hastalıklarda rolü olduğu bilinen BECN1, ATG5, mTOR geni ile yaşam uzunluğunu etkileyen Vg ve AmILP-2 gen ifade farklılıkları araştırılmış, istatistiksel olarak anlamlılıkları tespit edilmiştir. Deney ünitelerinin beslenmesinde kullanılan şuruplar %50 sukroz ve steril su ile hazırlanmıştır ayrıca aktivatör gruplar için 3 mg/l kurkumin, inhibitör gruplar için 1,1 g/l sodyum bütirat eklenmiştir. Kontrol grubuna yalnızca şekerli su verilmiştir. Kraliçe larvalar, işçi larvalar ve ergin arılardan oluşan 36 numunede RNA izolasyonu, cDNA sentezi ve qPZR ile gen ekspresyonlarına bakılmıştır. Gen ekspresyon seviyeleri ∆∆Cp, ANOVA test, Tukey Kramer post hoc testi ve Pearson korelasyon katsayısı istatistik yöntemleri ile hesaplanmıştır ve ifade anlamlılığı değerlendirilmiştir. Uygulanan besin diyeti ana arı larvalarında mTOR ve SIRT1, işçi arı larvalarında ATG5, ergin işçi arılardam TOR ve BRP genlerinde istatistiki fark önemli bulunmuştur. Besin diyetinden bağımsız olarak çalışmamızda larva grubunda, SIRT1 ve SIRT6 genleri ATG5 geni ile SIRT7 geni ise mTOR ve ILP2 genleri ile ergin işçi arı grubunda ise SIRT1 geni mTOR, ILP2, ATG5ve Vg genleri ile SIRT6 ve SIRT7 genleri ise mTOR, BECN1, ILP2, ATG5 ve Vg genleri ile anlamlı bir korelasyon göstermiştir. Sonuç olarak, bu tez çalışmasında kurkumin ve sodyum bütirat içeren besin diyetlerinin doğrudan ya da dolaylı olarak sirtuin genleri venöro dejeneratif hastalıklarla ilişki gen üzerindeki etkileri gösterilmiştir. Daha fazla örneklem grubu ve parametreler içeren gen ve etkilerinin protein düzeylerinde de araştırıldığı çalışmaların yapılmasının yararlı olacağı düşünülmekledir.
There is currently no cure for neurodegenerative diseases. Honey bees are used as a model organism in the research of many neurodegenerative disorders. The sirtuins, which are histone deacetylases, are known to contribute to neurodegenerative diseases but it is not known in which genes and pathways they are involved. Therefore, in this thesis study, the activities of SIRT1, SIRT6, and SIRT7 were investigated in honeybees, and expression of related genes including the BRP gene that controls neurotransmitter release, BECN1, ATG5, and mTOR genes, which are known for their roles in neurodegenerative diseases, Vg and AmILP2 genes, which affect life span, were investigated and their statistical significance was determined. The syrups used in the feeding of the test units were prepared with 50% sucrose and sterile water, to which were added 3 mg/l curcumin for activator groups, and 1,1 g/l sodium butyrate for inhibitor groups. The control group was given only sugar water without additives. In 36 samples of queen larvae, worker larvae and adult bees, gene expression levels were examined by RNA isolation, cDNA synthesis and qPCR. Gene expression levels were calculated using Δ∆Cp, and the significance of the expression levels assessed using ANOVA test, Tukey Kramer post hoc test and Pearson correlation coefficient statistical methods. Statistically significant differences were found between nutrient diet applications in mTOR and SIRT1 in the queen bee larvae, ATG5 in worker bee larvae, mTOR and BRP in adult worker bees. In our study, regardless of the nutrient diet, the SIRT1 and SIRT6 genes showed a significant correlation with the ATG5 gene and the SIRT7 gene showed a significant correlation with the mTOR and ILP2 genes in the larval group, the SIRT1 gene was significantly correlated with themTOR, ILP2, ATG5, and Vg genes, and the SIRT6 and SIRT7 genes, such as mTOR, BECN1, ILP2, ATG5, and Vg genes in the adult worker bee group. Our result, this thesis has shown the effects of food diets containing curcumin and sodium butyrate directly or indirectly on the expression of sirtuin genes and genes associated with neurodegenerative diseases. It is thought that it would be useful to carry out studies including more sample groups and parameters and in which genes and their effects are also investigated at the protein level.
There is currently no cure for neurodegenerative diseases. Honey bees are used as a model organism in the research of many neurodegenerative disorders. The sirtuins, which are histone deacetylases, are known to contribute to neurodegenerative diseases but it is not known in which genes and pathways they are involved. Therefore, in this thesis study, the activities of SIRT1, SIRT6, and SIRT7 were investigated in honeybees, and expression of related genes including the BRP gene that controls neurotransmitter release, BECN1, ATG5, and mTOR genes, which are known for their roles in neurodegenerative diseases, Vg and AmILP2 genes, which affect life span, were investigated and their statistical significance was determined. The syrups used in the feeding of the test units were prepared with 50% sucrose and sterile water, to which were added 3 mg/l curcumin for activator groups, and 1,1 g/l sodium butyrate for inhibitor groups. The control group was given only sugar water without additives. In 36 samples of queen larvae, worker larvae and adult bees, gene expression levels were examined by RNA isolation, cDNA synthesis and qPCR. Gene expression levels were calculated using Δ∆Cp, and the significance of the expression levels assessed using ANOVA test, Tukey Kramer post hoc test and Pearson correlation coefficient statistical methods. Statistically significant differences were found between nutrient diet applications in mTOR and SIRT1 in the queen bee larvae, ATG5 in worker bee larvae, mTOR and BRP in adult worker bees. In our study, regardless of the nutrient diet, the SIRT1 and SIRT6 genes showed a significant correlation with the ATG5 gene and the SIRT7 gene showed a significant correlation with the mTOR and ILP2 genes in the larval group, the SIRT1 gene was significantly correlated with themTOR, ILP2, ATG5, and Vg genes, and the SIRT6 and SIRT7 genes, such as mTOR, BECN1, ILP2, ATG5, and Vg genes in the adult worker bee group. Our result, this thesis has shown the effects of food diets containing curcumin and sodium butyrate directly or indirectly on the expression of sirtuin genes and genes associated with neurodegenerative diseases. It is thought that it would be useful to carry out studies including more sample groups and parameters and in which genes and their effects are also investigated at the protein level.
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Keywords
Bal arısı, Sirtuin, BECN1, ATG5, mTOR, BRP, Vg ve AmILP-2, HDAC inhibitör, HDAC aktivatör, Nörodejeneratif hastalıklar, Honey bee, Vg and AmILP2, HDAC activator, Neurodegenerative diseases
Citation
Çelik, S. (2024). Bal arılarında (Apis mellifera) beslenme farklılıklarının sirtuin enzimleri ve nörodejeneratif ve nörotransmitter salınımını düzenleyen genler üzerinde etkilerinin araştırılması. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Fen Bilimleri Enstitüsü.