Quercetin’in HepG2 hücrelerinde, TNF-Alfa ve LPS ile indüklenen enflamasyonda, eritropoietin, HIF-1α, HIF-2α ve piroptoz ile ilgili genlerin ekspresyon seviyeleri üzerine etkileri

Year 2023, Volume: 48 Issue: 1, 268 - 277, 31.03.2023

Mete Ozkurt , Burcu Akıncı , Selda Kabadere Deliorman , Rumeysa Özyurt , Abdullah Karadağ , Nilüfer Erkasap

https://doi.org/10.17826/cumj.1174607

Abstract

Amaç: Sistemik inflamasyon sonucu dolaşımda artan TNF-α, eritropoezi düzenleyici hormon eritropoietin (EPO) üretimini baskılayarak inflamasyon anemisine neden olur. Quercetin'in antiinflamatuar aktivitesi farklı çalışmalarda rapor edilmiş ancak bu durumun EPO gen ekspresyonu üzerine etkisini gösteren bir çalışmaya rastlanmamıştır. Bu amaçla quercetin'in inflamasyon ve daha da önemlisi inflamatuar ortamda baskılanan EPO ve EPO sentezini uyaran Hipoksi ile indüklenebilir faktörler (HIF) üzerindeki etkilerini değerlendirdik. Ayrıca proinflamatuvar programlı hücre ölümü olarak tanımlanan piroptozis üzerine quercetin'in etkisini değerlendirmeyi amaçladık.
Gereç ve Yöntem: İn-vitro HepG2 hücrelerini kullanarak TNF-α veya LPS ile inflamasyon modelleri oluşturduk. Bu inflamasyon modellerinde, quercetin'in proinflamatuar mediatörler TNF-α, IL-1α, NF-κB gen ekspresyonları, EPO, HIF-1α, HIF-2α gen ekspresyonları ve ayrıca piroptozla ilişkili kaspaz-1 ve IL-18 gen ifadesi üzerindeki etkilerini değerlendirdik..
Bulgular: Quercetin, TNF-α, IL-1α, NF-κB mRNA düzeylerini yükselterek inflamatuar etkiler göstermiştir. Bu enflamatuar etkiyle uyumlu olarak, EPO mRNA ekspresyonu baskılanmıştır. HIF-1α ve HIF-2α mRNA seviyeleri yükselmiştir.
Sonuç: HIF'lerdeki artışın, inflamatuar sitokinlerin ve NF-κB'nin EPO üretimi üzerindeki baskılayıcı etkisini engelleyemediğini düşündürmektedir. Ancak kaspaz-1 ve IL-18 mRNA düzeylerini düşürerek proinflamatuar hücre ölümünü baskılama eğiliminde olduğu gözlemlenmiştir. Bu sonuçlar, quercetin'in çelişkili etkiler gösterebileceğini ve güvenliğini test etmek için daha ileri çalışmalara ihtiyaç olduğunu göstermektedir.

Keywords

inflamasyon anemisi, eritropoetin, HİF, TNF-alfa, quercetin

Supporting Institution

Eskişehir Osmangazi Üniversitesi, Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

2018-2004

The effects of quercetin on erythropoietin, hypoxia-inducible factors-1α, 2α, and pyroptosis related genes expression levels in inflammation induced by TNF-α and LPS in HepG2 cells

Abstract

Purpose: TNF-α, which increases in the circulation as a result of systemic inflammation, suppresses the production of the erythropoiesis regulating hormone, erythropoietin (EPO) and causes inflammation anemia. The anti-inflammatory activity of quercetin has been reported in different studies. However, no study has been found showing the effect of this situation on EPO gene expression. For this purpose, we evaluated the effects of quercetin on inflammation and, more importantly, its effects on EPO, suppressed in the inflammatory environment and Hypoxia-inducible factors (HIF), which are stimulators of EPO synthesis. In addition, we aimed to evaluate the effect of quercetin on pyroptosis, which is defined as pro-inflammatory programmed cell death.
Materials and Methods: We created inflammation models with TNF-α or LPS using HepG2 cells in vitro. In these inflammation models, we evaluated the effects of quercetin on proinflammatory mediators TNF-α, IL-1α, NF-κB gene expressions, EPO, HIF-1α, HIF-2α gene expressions, as well as pyroptosis-related caspase 1 and IL-18 gene expressions.
Results: Quercetin showed inflammatory effects by increasing TNF-α, IL-1α, and NF-κB mRNA levels. Consistent with this inflammatory effect, EPO mRNA expression was suppressed. HIF-1α and HIF-2α mRNA levels were increased.
Conclusion: These results suggested that the increase in HIFs could not prevent the suppressive effect of inflammatory cytokines and NF-κB on EPO production. However, it has been observed that it tends to suppress proinflammatory cell death by decreasing caspase 1 and IL-18 mRNA levels. These results show that quercetin may show conflicting effects, and further studies are needed to test its safety.

Keywords

anemia of inflammation, erythropoietin, HIF, TNF-alpha, quercetin

Project Number

2018-2004

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