Publication: Kan nötrofil seviyesi değişikliğinin total kolesterol düzeyi üzerine etkisinde karaciğer makrofaj ve ATP-bağlayıcı-kaset-taşıyıcılarının rolü
Abstract
Amaç: Kolesterol, nötrofiller için hücre membranında bulunan önemli bir bileşen olarak kabul edilir. Bununla birlikte, ekstraselüler kolesterolün endositoz yoluyla nötrofil membranına entegre olma kabiliyeti ve kolesterol yüzey değişimi ile bu hücrelerin işlevselliğine etki edebilme potansiyeli bulunmaktadır. Kolesterol ve nötrofiller arasındaki ilişki inflamatuar açıdan ele alınmış olup nötrofillerin sayısında ya da fenotipinde meydana gelen değişimlerin kolesterol düzeyi üzerine olan etkisi ile halen bilinmemektedir. Gereç ve Yöntem: Bu bilgiler ışığında bu tez çalışmasında anti-nötrofil serum uygulanması ile nötropenik hale getirilen farelerin kan kolesterol düzeyleri ölçülmüş ve hemogram analizleri yapılmıştır. Bu işlemler tamamlandıktan sonra karaciğerden MACS-tabanlı yöntem ile F4-80+ hücreler izole edilmiş ve bu hücrelerin ATP-Bağlayıcı-Kaset-Taşıyıcı-1 (ABCA1) proteinleri akış-sitometrisi ile değerlendirilmiştir. Bulgular: Anti-nötrofil-serum (ANS) alan grupta kontrol grubu ile kıyaslandığında nötrofil seviyeleri istatiksel olarak anlamlı ölçüde azalmış (p<0.05), total lökosit, lenfosit ve monosit miktarları değişmemiştir. Düşük yoğunluklu lipoprotein (LDL) kolesterol miktarı ANS alan grupta (p<0.005) kontrol grubuna kıyasla anlamlı ölçüde artmıştır. Karaciğer hücrelerinden izole edilen F4-80+ hücrelerin sayısı kontrol ve ANS gruplarında anlamlı seviyede farklılık göstermezken, bu hücrelerin eksprese ettiği ABCA1 protein miktarı ANS grubunda anlamlı seviyede artmıştır (p<0.001). Sonuç: Dolaşımdaki nötrofil seviyelerinin azalması LDL kolesterol düzeyini artırmış, bu artıştan karaciğerden izole edilen F4-80+ hücrelerin eksprese ettiği ABCA1 proteinler sorumludur
Aim: Cholesterol is considered an important component of the cell membrane for neutrophils. Extracellular cholesterol has the ability to integrate into the neutrophil membrane through endocytosis and has the potential to affect the functionality of these cells through cholesterol surface modification. The relationship between cholesterol and neutrophils has been discussed from an inflammatory perspective, but the effect of changes in the number or phenotype of neutrophils on cholesterol levels is still unknown. Material and Method: In the light of this information, in this thesis study, blood cholesterol levels of mice made neutropenic by the application of anti-neutrophil serum were measured and hemogram analyzes were performed. After these procedures were completed, F4-80+ cells were isolated from the liver by the MACS-based method, and the ATP-Binding-Cassette-Carrier-1 (ABCA1) proteins of these cells were evaluated by flow-cytometry. Result: In the group receiving anti-neutrophil-serum (ANS), compared to the control group, neutrophil levels decreased statistically significantly (p<0.05), and total leukocyte, lymphocyte and monocyte amounts did not change. The amount of low density lipoprotein (LDL) cholesterol increased significantly in the group receiving ANS (p<0.005) compared to the control group. While the number of F4-80+ cells isolated from liver cells did not differ significantly in the control and ANS groups, the amount of ABCA1 protein expressed by these cells increased significantly in the ANS group (p<0.001). Conclusion: The decrease in circulating neutrophil levels increased the LDL cholesterol level, and ABCA1 proteins expressed by F4-80+ cells isolated from the liver are responsible for this increase.
Aim: Cholesterol is considered an important component of the cell membrane for neutrophils. Extracellular cholesterol has the ability to integrate into the neutrophil membrane through endocytosis and has the potential to affect the functionality of these cells through cholesterol surface modification. The relationship between cholesterol and neutrophils has been discussed from an inflammatory perspective, but the effect of changes in the number or phenotype of neutrophils on cholesterol levels is still unknown. Material and Method: In the light of this information, in this thesis study, blood cholesterol levels of mice made neutropenic by the application of anti-neutrophil serum were measured and hemogram analyzes were performed. After these procedures were completed, F4-80+ cells were isolated from the liver by the MACS-based method, and the ATP-Binding-Cassette-Carrier-1 (ABCA1) proteins of these cells were evaluated by flow-cytometry. Result: In the group receiving anti-neutrophil-serum (ANS), compared to the control group, neutrophil levels decreased statistically significantly (p<0.05), and total leukocyte, lymphocyte and monocyte amounts did not change. The amount of low density lipoprotein (LDL) cholesterol increased significantly in the group receiving ANS (p<0.005) compared to the control group. While the number of F4-80+ cells isolated from liver cells did not differ significantly in the control and ANS groups, the amount of ABCA1 protein expressed by these cells increased significantly in the ANS group (p<0.001). Conclusion: The decrease in circulating neutrophil levels increased the LDL cholesterol level, and ABCA1 proteins expressed by F4-80+ cells isolated from the liver are responsible for this increase.