Kafein akrilamid uygulanan insan kaynaklı hepatoma hücrelerinde protein oksidasyonu ve endoplazmik retikulum stresine etkisi
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Dosyalar
Tarih
2020
Yazarlar
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Yayıncı
Sosyal Bilimler Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Çalışmamızın amacı, kafeinin akrilamid uygulanan insan kaynaklı hepatoma hücrelerinde protein oksidasyonu ve endoplazmik retikulum stresine etkisini araştırmaktır. Akrilamid ve akrilamid ile birlikte uygulanan kafeinin; hücre canlılığına etkisi 3-(4,5- dimetil-2-tiyazolil)-2,5-difenil-2H-tetrazolyum bromür testi ile, protein karbonil, glukozla ilişkili protein 78, transkripsiyon aktive edici faktör 4, C/EBP-homolog protein düzeyleri western blot yöntemiyle ölçüldü. HepG2 hücrelerine 24 saat süreyle uygulanan 1000 ve 10000 ?M konsantrasyonlarında akrilamid hücre canlılığını anlamlı olarak azalttı (her ikisi için p<0.05). 10000 ?M akrilamid, ER stres belirteci olan glukozla ilişkili protein 78, transkripsiyon aktive edici faktör 4, C/EBP-homolog protein düzeylerini anlamlı olarak arttırdı (tümü için p<0.05). Kafein uygulanan hücrelerde glukozla ilişkili protein 78, transkripsiyon aktive edici faktör 4, C/EBP-homolog protein düzeyleri anlamlı olarak değişmedi (tümü için p>0.05). Protein karbonil düzeyleri 24 saat süre ile 10000 ?M akrilamid uygulanan hücrelerde kontrole göre anlamlı oranda yüksekti. 50 ve 200 ?M kafein, 10000 ?M akrilamid uygulanan hücrelerde protein karbonil düzeylerini anlamlı olarak önledi (ikisi için p<0.05). Sonuç olarak çalışmamız, HepG2 hücrelerinde akrilamidin 1000 ve 10000 ?M konsantrasyonlarında hücre canlılığını azalttığını, protein oksidasyonu ve endoplazmik retikulum stresi arttırdığını, 10000 ?M akrilamid ile birlikte uygulanan 10, 50 ve 200 ?M kafeinin endoplazmik retikulum stresi değiştirmediğini ancak 50 ve 200 ?M konsantrasyonlarında kafeinin protein oksidasyonunu önlediğini göstermiştir.
Our study aims to investigate the effect of caffeine on protein oxidation and endoplasmic reticulum stress in human-induced hepatoma cells treated with acrylamide. The effect of caffeine administered with acrylamide and acrylamide on cell viability was measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, protein carbonyl, glucose regulated protein 78, activating transcription factor 4, C/EBPhomologous protein levels were measured by western blot method. Applied for 24 hours, 1000 and 10000 ?M acrylamide significantly reduced the viability of HepG2 cells (for both p <0.05). 10000 ?M acrylamide significantly increased the protein levels of glucose regulated protein 78, activating transcription factor 4, C/EBPhomologous protein (p<0.05 for all). The protein levels of glucose regulated protein 78, activating transcription factor 4, C/EBP-homologous protein were not significantly changed in 10, 50, 200 ?M caffeine treated cells (p >0.05 for all). The protein carbonyl levels were significantly higher in cells treated with 10000 ?M acrylamide compared to control. 50 and 200 ?M caffeine significantly prevented protein carbonyl levels in cells treated with 10000 ?M acrylamide (p <0.05 for both). As a result, our study showed that acrylamide decreases cell viability at concentrations of 1000 and 10000 ?M in HepG2 cells, increases protein oxidation and endoplasmic reticulum stress, caffeine did not change endoplasmic reticulum stress, but doses of 50 and 200 ?M caffeine prevent protein oxidation
Our study aims to investigate the effect of caffeine on protein oxidation and endoplasmic reticulum stress in human-induced hepatoma cells treated with acrylamide. The effect of caffeine administered with acrylamide and acrylamide on cell viability was measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, protein carbonyl, glucose regulated protein 78, activating transcription factor 4, C/EBPhomologous protein levels were measured by western blot method. Applied for 24 hours, 1000 and 10000 ?M acrylamide significantly reduced the viability of HepG2 cells (for both p <0.05). 10000 ?M acrylamide significantly increased the protein levels of glucose regulated protein 78, activating transcription factor 4, C/EBPhomologous protein (p<0.05 for all). The protein levels of glucose regulated protein 78, activating transcription factor 4, C/EBP-homologous protein were not significantly changed in 10, 50, 200 ?M caffeine treated cells (p >0.05 for all). The protein carbonyl levels were significantly higher in cells treated with 10000 ?M acrylamide compared to control. 50 and 200 ?M caffeine significantly prevented protein carbonyl levels in cells treated with 10000 ?M acrylamide (p <0.05 for both). As a result, our study showed that acrylamide decreases cell viability at concentrations of 1000 and 10000 ?M in HepG2 cells, increases protein oxidation and endoplasmic reticulum stress, caffeine did not change endoplasmic reticulum stress, but doses of 50 and 200 ?M caffeine prevent protein oxidation
Açıklama
Anahtar Kelimeler
Akrilamid, Kafein, HepG2, Protein karbonil, Endoplazmik retikulum stresi, Acrylamide, Caffeine, HepG2, Protein carbonyl, Endoplasmic reticulum stress
