İn vitro hidrojen peroksit uyarımlı karaciğer oksidatif stres modelinde 3-(4-hidroksifenil) propiyonik asidin etkinliğinin araştırılması
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Tarih
2021
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Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Polifenollerin insan mikrobiyota ürünlerinin antioksidan özellikleri birçok araştırmada ortaya konulmuştur. Karaciğer temel metabolizan organ olduğu için oksidatif strese yoğun olarak maruz kalmaktadır. Araştırmanın amacını birçok çalışmada oksidan molekül olan hidrojen peroksitin (H2O2) karaciğer hücrelerine uygulanımı sonrası oluşturulan in vitro oksidatif stres modelinde, bir mikrobiyota metaboliti olan 3-(4-hidroksifenil) propiyonik asit (4-HPPA)'in antioksidan kapasitesinin ortaya konulması oluşturmaktadır. Araştırmada insan orijinli karaciğer HepG2 hücreleri materyal olarak kullanıldı. Deneme modelllemesinde hidrojen peroksit ve 4-HPPA temin edilerek ve farklı konsantrasyonlarda karaciğer hücrelerine 1 gün süreyle uygulandı. Bu maddelerin etkin miktarları, canlılık analizleriyle tespit edildi. Deneme süreleri sonunda gruplardan alınan RNA örneklerinden cDNA dönüşümleri gerçekleştirilerek; süperoksit dismutaz (SOD), glutatyon peroksidaz (GSH-Px) ve katalaz (CAT) gen ekspresyon düzeyleri gerçek zamanlı kuantatif polimeraz zincir reaksiyonlarıyla (QRT-PCR) ortaya konuldu. Çalışmadan elde edilen verilere göre H2O2'in farklı konsantrasyonları hücrelere uygulanarak kontrol grubuna göre %23 hücre kayıpları meydana getiren 100 μM'lık konsantrasyonu etkin olarak tespit edildi. Bununla birlikte 4-HPPA'nın 25 μg/ml bu kayıpları %17 oranında önlediği tespit edildi. Bu bulgulara göre gen ekspresyon analizleri bu konsantrasyonlar kullanılarak hücrelere uygulandı. Gen ekspresyon analizleri incelendiğinde hidrojen peroksitin 1 gün süreli uygulanımı GSH-Px ve CAT ekspresyonlarını relatif misli değerlerde baskılarken, 4-HPPA'nın bu olumsuz tabloyu önemli derecede geri çevirdiği tespit edildi. Bununla birlikte konu ile ilgili literatür incelendiğinde SOD sonuçlarının doz ve zamana bağlı olarak değişkenlik gösterebileceği görüldü. Sonuç olarak çalışmadan elde edilen bulgulara göre, 4-HPPA'nın oksidatif stres kaynaklı akut karaciğer hasarında tedaviye katkı sağlayabileceği tespit edildi. Ancak elde edilen bulgular ile tam olarak etkinliğinin söylenebilmesi için in vivo denemelere ve ileri moleküler yöntemlere ihtiyaç duyulmaktadır.
The antioxidant properties of human microbiota products of polyphenols have been demonstrated in many studies. Since the liver is the main metabolizing organ, it is heavily exposed to oxidative stress. The aim of this study is to demonstrate the antioxidant capacity of 3-(4-hydroxyphenyl) propionic acid (4-HPPA), a microbiota metabolite, in an in vitro oxidative stress model created after the application of the oxidant molecule hydrogen peroxide (H2O2) to liver cells. Liver HepG2 cells of human origin from continuous cell lines were used as material in the experiment. In trial modelling, hydrogen peroxide and 4-HPPA were provided and applied to liver cells at different concentrations for 1 day. The effective concentrations of these substances were determined by viability tests. At the end of the trial period, cDNA conversions were performed from the RNA samples taken from the groups and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) catalase (CAT) gene expression levels were determined by real-time quantitative polymerase chain reactions (QRT-PCR). According to the data obtained from the study, different concentrations of H2O2 were applied to the cells and the concentration of 100 μM, which caused 23% cell loss compared to the control group, was determined as the effective concentration. However, it was determined that 25 μg/ml 4-HPPA prevented these losses by 17%. According to these findings, gene expression analyzes were performed on cells using these concentrations. When the gene expression analyzes were examined, it was determined that the 1-day application of hydrogen peroxide suppressed the GSH-Px and CAT expressions at relative values, while 4-HPPA significantly reversed this negative picture. However, when the literature on the subject was examined, it was seen that the results of SOD could vary depending on the dose and time. As a result, according to the findings obtained from the study, it was determined that 4-HPPA may contribute to the treatment of acute liver injury caused by oxidative stress. However, in vivo trials and advanced molecular methods are needed to fully express its effectiveness with the findings obtained.
The antioxidant properties of human microbiota products of polyphenols have been demonstrated in many studies. Since the liver is the main metabolizing organ, it is heavily exposed to oxidative stress. The aim of this study is to demonstrate the antioxidant capacity of 3-(4-hydroxyphenyl) propionic acid (4-HPPA), a microbiota metabolite, in an in vitro oxidative stress model created after the application of the oxidant molecule hydrogen peroxide (H2O2) to liver cells. Liver HepG2 cells of human origin from continuous cell lines were used as material in the experiment. In trial modelling, hydrogen peroxide and 4-HPPA were provided and applied to liver cells at different concentrations for 1 day. The effective concentrations of these substances were determined by viability tests. At the end of the trial period, cDNA conversions were performed from the RNA samples taken from the groups and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) catalase (CAT) gene expression levels were determined by real-time quantitative polymerase chain reactions (QRT-PCR). According to the data obtained from the study, different concentrations of H2O2 were applied to the cells and the concentration of 100 μM, which caused 23% cell loss compared to the control group, was determined as the effective concentration. However, it was determined that 25 μg/ml 4-HPPA prevented these losses by 17%. According to these findings, gene expression analyzes were performed on cells using these concentrations. When the gene expression analyzes were examined, it was determined that the 1-day application of hydrogen peroxide suppressed the GSH-Px and CAT expressions at relative values, while 4-HPPA significantly reversed this negative picture. However, when the literature on the subject was examined, it was seen that the results of SOD could vary depending on the dose and time. As a result, according to the findings obtained from the study, it was determined that 4-HPPA may contribute to the treatment of acute liver injury caused by oxidative stress. However, in vivo trials and advanced molecular methods are needed to fully express its effectiveness with the findings obtained.
Açıklama
Anahtar Kelimeler
Biyokimya, Biochemistry
