The Effect of Glutathione on Necrotic and Autophagic Pathways in Nrk-52e Cell Line Administered With Cytotoxic Concentration of Glucose.
Abstract
Bu tez çalışmasında sitotoksik konsantrasyonda glukoz ile diyabetik stres oluşturulan böbrek hücre hattında, diyabete bağlı komplikasyonların ortaya konulmasında önemli olan prooksidan/antioksidan dengenin glutatyon uygulanmasından nekrotik ve otofajik yolakların nasıl etkilendiğini araştırmak ve elde edilen sonuçlara göre, komplikasyonların giderilmesine glutatyonun önemli bir etkisinin olup olmadığını ortaya koymak amacıyla planlandı.NRK-52E hücresi in-vitro koşullarında çoğaltıldı. Glukozun IC50 değeri ve glutatyonun proliferatif dozunu belirlemek için MTT canlılık testi yapıldı. Hücre hattında çalışma grupları konrol, yüksek glukoz, glutatyon ve yüksek glukoz +glutatyon olarak belirlendi. 24 saat inkübasyondan sonra hücreler toplandı. Trizol ile RNA eldesi yapıldı. cDNA sentezi yapılarak nekrotik ve otofajik genlerin ekspresyon seviyeleri için RT-qPCR analizi ile yapıldı. Bu analiz sonucuna göre; otofajik ve nekrotik genlerin glutatyon grubunda değişmediği, glukoz uygulan gruplarda up-regüle olduğu glutatyon uygulaması ile azalarak kontrole yaklaştığı tespit edildi. Anahtar kelimeler: Yüksek glukoz, Hücre kültürü, Glutatyon, Nekrozis, Otofaji
In this thesis study, diabetic stress was induced by glucose at cytotoxic concentration. It was planned to investigate how necrotic and autophagic pathways are affected by glutathione administration on the prooxidant/antioxidant balance, which is important in revealing diabetes-related complications in the kidney cell line, and to reveal whether glutathione has a significant effect on the elimination of complications according to the results obtained. reproduced under the conditions. MTT viability test was performed to determine the IC50 value of glucose and the proliferative dose of glutathione. In the cell line, the study groups were determined as control, high glucose, glutathione and high glucose + glutathione. After 24 h of incubation, cells were harvested. RNA was obtained with Trizol. cDNA was made and expression levels of necrotic and autophagic genes were performed by RT-qPCR. According to the result of this analysis; It was determined that autophagic and necrotic genes did not change in the glutathione group, were up-regulated in the glucose-administered groups, and decreased with glutathione administration and approached control. Key words: High glucose, Cell culture, Glutathione, Necrosis, Autophagy
In this thesis study, diabetic stress was induced by glucose at cytotoxic concentration. It was planned to investigate how necrotic and autophagic pathways are affected by glutathione administration on the prooxidant/antioxidant balance, which is important in revealing diabetes-related complications in the kidney cell line, and to reveal whether glutathione has a significant effect on the elimination of complications according to the results obtained. reproduced under the conditions. MTT viability test was performed to determine the IC50 value of glucose and the proliferative dose of glutathione. In the cell line, the study groups were determined as control, high glucose, glutathione and high glucose + glutathione. After 24 h of incubation, cells were harvested. RNA was obtained with Trizol. cDNA was made and expression levels of necrotic and autophagic genes were performed by RT-qPCR. According to the result of this analysis; It was determined that autophagic and necrotic genes did not change in the glutathione group, were up-regulated in the glucose-administered groups, and decreased with glutathione administration and approached control. Key words: High glucose, Cell culture, Glutathione, Necrosis, Autophagy
Description
Keywords
Biyokimya, Glükoz, Glütatyon, Hücre dizisi, Hücre kültür teknikleri, Nekroz, Otofaji, Sitotoksisite, Biochemistry, Glucose, Glutathione, Cell line, Cell culture techniques, Necrosis, Autophagy, Cytotoxicity
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