Borik asitin NFKB sinyali iletim yolağına etkisinin NFKB-GFP belirteç sistemi entegre edilmiş transgenic HEK293 hücre hattı kullanılarak çalışılması
Küçük Resim Yok
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Araştırma, insan sağlığı üzerine birçok metabolik ve biyokimyasal etkisi olduğu bilinen borik asitin, inflamatuar yanıtların anahtar düzenleyicisi olan NF-?B yolağını regüle edebilme potansiyelini belirlemek amacıyla yapılmıştır. Materyal ve Metot: Bu araştırmada NF-?B-GFP belirteç sistemi entegre edilmiş transgenik HEK293 hücreleri model olarak kullanılmıştır. Bu doğrultuda, çalışma için uygun TNF? ve borik asit miktarları tespit edilmiştir. Ardından TNF? ile uyarılmış ve aktifleşmiş NF-?B sinyal iletim yolağına, kontrol grubuna kıyasla borik asitin etkisi incelenmiştir. Borik asitin NF-?B yolağını regüle edebilme kapasitesi akış sitometrik analizlerle belirlenmiştir. TNF? ve borik asit uygulanmış hücrelerin GFP yoğunluğunun ortalaması ve standart sapması, üç bağımsız biyolojik deneyden hesaplanmıştır. İstatistiksel testler için iki örneklem konum t-testi kullanılmıştır. Bulgular: Bu çalışmada kullanılan HEK293 hücrelerinin genomuna entegre GFP kaseti bulunduğundan, akış sitometrik analizlerle kontrol grubunda bir miktar GFP ekspresyonu gözlemlenmiştir. Belirli dozlarda TNF? verildiğinde, hücrelerde kontrol grubuna kıyasla 6-8 kat daha yoğun GFP ekspresyonu belirlenirken (p<0.0001), sadece borik asit uygulandığında, hücrelerde kontrol grubuna kıyasla eşit veya daha az yoğunlukta GFP ekspresyonu gözlemlenmiştir (p<0.05). TNF? ve farklı dozlarda borik asit birlikte verildiğinde ise, TNF? grubuna kıyasla GFP yoğunluğunun azaldığı gözlemlenmiştir (p<0.05). Sonuç: Analizler sonucunda, HEK293 hücrelerine tek başına borik asitin muamelesi kontrole kıyasla etkili bir fark oluşturmamışken, TNF? ile aktive edilen yolakta borik asit muamelesinin NF-?B aktivasyonunu azaltıcı yönde etkisi olduğu ortaya konmuştur.
Aim: The research was carried out to determine the potential of boric acid, which is known to have many metabolic and biochemical effects on human health, to regulate the NF-kB pathway, which is a key regulator of inflammatory responses. Material and Method: In this study, transgenic HEK293 cells with integrated NF-?B-GFP marker system were used as a model. In this direction, suitable amounts of TNF? and boric acid were determined for the study. Then, the effect of boric acid on TNF? stimulated and activated NF-kB signal transduction pathway compared to the control group was investigated. The capacity of boric acid to regulate the NF-kB pathway was determined by flow cytometric analysis. The mean and standard deviation of the GFP density of TNF? and boric acid-treated cells were calculated from three independent biological experiments. Two-sample position t-test was used for statistical tests. Results: Since the HEK293 cells used in this study had a GFP cassette integrated into their genome, some GFP expression was observed in the control group by flow cytometric analysis. When given specific doses of TNF?, 6-8 times more intense GFP expression was determined in the cells compared to the control group (p<0.0001), while when only boric acid was applied, equal or less intense GFP expression was observed in the cells compared to the control group (p<0.05). When TNF? and different doses of boric acid were given together, it was observed that the GFP density decreased compared to the TNF? group (p<0.05). Conclusion: As a result of the analysis, it was revealed that boric acid treatment in HEK293 cells alone did not make an effective difference compared to the control, while boric acid treatment had a decreasing effect on NF-kB activation in the pathway activated by TNF?.
Aim: The research was carried out to determine the potential of boric acid, which is known to have many metabolic and biochemical effects on human health, to regulate the NF-kB pathway, which is a key regulator of inflammatory responses. Material and Method: In this study, transgenic HEK293 cells with integrated NF-?B-GFP marker system were used as a model. In this direction, suitable amounts of TNF? and boric acid were determined for the study. Then, the effect of boric acid on TNF? stimulated and activated NF-kB signal transduction pathway compared to the control group was investigated. The capacity of boric acid to regulate the NF-kB pathway was determined by flow cytometric analysis. The mean and standard deviation of the GFP density of TNF? and boric acid-treated cells were calculated from three independent biological experiments. Two-sample position t-test was used for statistical tests. Results: Since the HEK293 cells used in this study had a GFP cassette integrated into their genome, some GFP expression was observed in the control group by flow cytometric analysis. When given specific doses of TNF?, 6-8 times more intense GFP expression was determined in the cells compared to the control group (p<0.0001), while when only boric acid was applied, equal or less intense GFP expression was observed in the cells compared to the control group (p<0.05). When TNF? and different doses of boric acid were given together, it was observed that the GFP density decreased compared to the TNF? group (p<0.05). Conclusion: As a result of the analysis, it was revealed that boric acid treatment in HEK293 cells alone did not make an effective difference compared to the control, while boric acid treatment had a decreasing effect on NF-kB activation in the pathway activated by TNF?.
Açıklama
Anahtar Kelimeler
Genetik, Genetics, Tıbbi Biyoloji
