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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
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.

Supporting Institution

Eskişehir Osmangazi Üniversitesi, Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

2018-2004

References

  • Weiss G, Ganz T, Goodnough LT. Anemia of inflammation. Blood. 2019;133:40-50.
  • Weiss G, Goodnough LT. Anemia of chronic disease. N Engl J Med. 2005;352:1011-23.
  • Jelkmann W. Regulation of erythropoietin production. J Physiol. 2011;589:1251-8.
  • Warnecke C, Zaborowska Z, Kurreck J, Erdmann VA, Frei U, Wiesener M et al. Differentiating the functional role of hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha (EPAS-1) by the use of RNA interference: erythropoietin is a HIF-2alpha target gene in Hep3B and Kelly cells. FASEB J. 2004;18:1462-4.
  • La Ferla K, Reimann C, Jelkmann W, Hellwig-Burgel T. Inhibition of erythropoietin gene expression signaling involves the transcription factors GATA-2 and NF-kappaB. FASEB J. 2002;16:1811-3.
  • Azeem M, Hanif M, Mahmood K, Ameer N, Chughtai FRS, Abid U. An insight into anticancer, antioxidant, antimicrobial, antidiabetic and anti-inflammatory effects of quercetin: a review. Polym Bull (Berl). 2023;80:241-62.
  • Nishimura K, Matsumoto R, Yonezawa Y, Nakagawa H. Effect of quercetin on cell protection via erythropoietin and cell injury of HepG2 cells. Arch Biochem Biophys. 2017;63:11-6.
  • Kalemci O, Aydin HE, Kizmazoglu C, Kaya I, Yilmaz H, Arda NM. Effects of quercetin and mannitol on erythropoietin levels in rats following acute severe traumatic brain injury. J Korean Neurosurg Soc. 2017;60:355-61.
  • Ozkurt M, Hellwig-Burgel T, Depping R, Kadabere S, Ozyurt R, Karadag A et al. miR663 Prevents Epo inhibition caused by TNF-alpha in normoxia and hypoxia. Int J Endocrinol. 2021;2021:3670499.
  • Hsiao CC, Chen PH, Cheng CI, Tsai MS, Chang CY, Lu SC et al. Toll-like receptor-4 is a target for suppression of proliferation and chemoresistance in HepG2 hepatoblastoma cells. Cancer Lett. 2015;368:144-52.
  • Jelkmann W, Hellwig-Buergel T. Tumor necrosis factor p55 receptor (TNF-RI) mediates the in vitro inhibition of hepatic erythropoietin production. Exp Hematol. 1999;27:224-8.
  • Li Y, Yao J, Han C, Yang J, Chaudhry MT, Wang S et al. Quercetin, inflammation and immunity. Nutrients. 2016;8:167.
  • Balstad TR, Paur I, Poulsen M, Markowski J, Kolodziejczyk K, Dragsted LO et al. Apple, cherry, and blackcurrant increases nuclear factor kappa B activation in liver of transgenic mice. Nutr Cancer. 2010;62:841-8.
  • Pang JL, Ricupero DA, Huang S, Fatma N, Singh DP, Romero JR et al. Differential activity of kaempferol and quercetin in attenuating tumor necrosis factor receptor family signaling in bone cells. Biochem Pharmacol. 2006;71:818-26.
  • Rangan GK, Wang Y, Harris DC. Dietary quercetin augments activator protein-1 and does not reduce nuclear factor-kappa B in the renal cortex of rats with established chronic glomerular disease. Nephron. 2002;90:313-9.
  • Granado-Serrano AB, Martin MA, Bravo L, Goya L, Ramos S. Quercetin attenuates TNF-induced inflammation in hepatic cells by inhibiting the NF-kappaB pathway. Nutr Cancer. 2012;64:588-98.
  • Jeon H, Kim H, Choi D, Kim D, Park SY, Kim YJ et al. Quercetin activates an angiogenic pathway, hypoxia inducible factor (HIF)-1-vascular endothelial growth factor, by inhibiting HIF-prolyl hydroxylase: a structural analysis of quercetin for inhibiting HIF-prolyl hydroxylase. Mol Pharmacol. 2007;71:1676-84.
  • Hellwig-Burgel T, Rutkowski K, Metzen E, Fandrey J, Jelkmann W. Interleukin-1beta and tumor necrosis factor-alpha stimulate DNA binding of hypoxia-inducible factor-1. Blood. 1999;94:1561-7.
  • Belaiba RS, Bonello S, Zahringer C, Schmidt S, Hess J, Kietzmann T et al. Hypoxia up-regulates hypoxia-inducible factor-1alpha transcription by involving phosphatidylinositol 3-kinase and nuclear factor kappaB in pulmonary artery smooth muscle cells. Mol Biol Cell. 2007;18:4691-7.
  • Ke S, Rabson AB, Germino JF, Gallo MA, Tian Y. Mechanism of suppression of cytochrome P-450 1A1 expression by tumor necrosis factor-alpha and lipopolysaccharide. J Biol Chem. 2001;276:39638-44.
  • Gires O, Kieu C, Fix P, Schmitt B, Munz M, Wollenberg B et al. Tumor necrosis factor alpha negatively regulates the expression of the carcinoma-associated antigen epithelial cell adhesion molecule. Cancer. 2001;92:620-8.
  • Krajewski J, Batmunkh C, Jelkmann W, Hellwig-Burgel T. Interleukin-1beta inhibits the hypoxic inducibility of the erythropoietin enhancer by suppressing hepatocyte nuclear factor-4alpha. Cell Mol Life Sci. 2007;64:989-98.
  • Bunn HF, Gu J, Huang LE, Park JW, Zhu H. Erythropoietin: a model system for studying oxygen-dependent gene regulation. J Exp Biol. 1998;201:1197-201.
  • McGettrick AF, O'Neill LAJ. The Role of HIF in immunity and inflammation. Cell Metab. 2020;32:524-36.
  • Yi YS. Regulatory roles of flavonoids on inflammasome activation during inflammatory responses. Mol Nutr Food Res. 2018;62:e1800147.
  • Malik A, Kanneganti TD. Inflammasome activation and assembly at a glance. J Cell Sci. 2017;130:3955-63.
  • Zhang W, Jia L, Zhao B, Xiong Y, Wang YN, Liang J et al. Quercetin reverses TNF alpha induced osteogenic damage to human periodontal ligament stem cells by suppressing the NFkappaB/NLRP3 inflammasome pathway. Int J Mol Med. 2021;47:39.
  • Choe JY, Kim SK. Quercetin and ascorbic acid suppress fructose-induced NLRP3 inflammasome activation by blocking intracellular shuttling of txnip in human macrophage cell lines. Inflammation. 2017;40:980-94.
  • Xue Y, Du M, Zhu MJ. Quercetin suppresses NLRP3 inflammasome activation in epithelial cells triggered by Escherichia coli O157:H7. Free Radic Biol Med. 2017;10:760-9.
  • Reyes-Farias M, Carrasco-Pozo C. The anti-cancer effect of quercetin: molecular implications in cancer metabolism. Int J Mol Sci. 2019;20:3177.
  • Jana N, Bretislav G, Pavel S, Pavla U. Potential of the flavonoid quercetin to prevent and treat cancer - current status of research. Klin Onkol. 2018;31:184-190.
  • Kabadere S, Oztopcu-Vatan P, Uyar R, Durmaz R. Quercetin both partially attenuates hydrogen peroxide-induced toxicity and decreases viability of rat glial cells. Acta Biol Hung. 2011;62:221-7.
  • Oztopcu-Vatan P, Kabadere S, Uyar R. The effect of pretreatment or combined treatment of quercetin on menadione toxicity in rat primary mixed glial cells in vitro. Cytotechnology. 2009;61:11-6

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

Year 2023, Volume: 48 Issue: 1, 268 - 277, 31.03.2023
https://doi.org/10.17826/cumj.1174607

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.

Project Number

2018-2004

References

  • Weiss G, Ganz T, Goodnough LT. Anemia of inflammation. Blood. 2019;133:40-50.
  • Weiss G, Goodnough LT. Anemia of chronic disease. N Engl J Med. 2005;352:1011-23.
  • Jelkmann W. Regulation of erythropoietin production. J Physiol. 2011;589:1251-8.
  • Warnecke C, Zaborowska Z, Kurreck J, Erdmann VA, Frei U, Wiesener M et al. Differentiating the functional role of hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha (EPAS-1) by the use of RNA interference: erythropoietin is a HIF-2alpha target gene in Hep3B and Kelly cells. FASEB J. 2004;18:1462-4.
  • La Ferla K, Reimann C, Jelkmann W, Hellwig-Burgel T. Inhibition of erythropoietin gene expression signaling involves the transcription factors GATA-2 and NF-kappaB. FASEB J. 2002;16:1811-3.
  • Azeem M, Hanif M, Mahmood K, Ameer N, Chughtai FRS, Abid U. An insight into anticancer, antioxidant, antimicrobial, antidiabetic and anti-inflammatory effects of quercetin: a review. Polym Bull (Berl). 2023;80:241-62.
  • Nishimura K, Matsumoto R, Yonezawa Y, Nakagawa H. Effect of quercetin on cell protection via erythropoietin and cell injury of HepG2 cells. Arch Biochem Biophys. 2017;63:11-6.
  • Kalemci O, Aydin HE, Kizmazoglu C, Kaya I, Yilmaz H, Arda NM. Effects of quercetin and mannitol on erythropoietin levels in rats following acute severe traumatic brain injury. J Korean Neurosurg Soc. 2017;60:355-61.
  • Ozkurt M, Hellwig-Burgel T, Depping R, Kadabere S, Ozyurt R, Karadag A et al. miR663 Prevents Epo inhibition caused by TNF-alpha in normoxia and hypoxia. Int J Endocrinol. 2021;2021:3670499.
  • Hsiao CC, Chen PH, Cheng CI, Tsai MS, Chang CY, Lu SC et al. Toll-like receptor-4 is a target for suppression of proliferation and chemoresistance in HepG2 hepatoblastoma cells. Cancer Lett. 2015;368:144-52.
  • Jelkmann W, Hellwig-Buergel T. Tumor necrosis factor p55 receptor (TNF-RI) mediates the in vitro inhibition of hepatic erythropoietin production. Exp Hematol. 1999;27:224-8.
  • Li Y, Yao J, Han C, Yang J, Chaudhry MT, Wang S et al. Quercetin, inflammation and immunity. Nutrients. 2016;8:167.
  • Balstad TR, Paur I, Poulsen M, Markowski J, Kolodziejczyk K, Dragsted LO et al. Apple, cherry, and blackcurrant increases nuclear factor kappa B activation in liver of transgenic mice. Nutr Cancer. 2010;62:841-8.
  • Pang JL, Ricupero DA, Huang S, Fatma N, Singh DP, Romero JR et al. Differential activity of kaempferol and quercetin in attenuating tumor necrosis factor receptor family signaling in bone cells. Biochem Pharmacol. 2006;71:818-26.
  • Rangan GK, Wang Y, Harris DC. Dietary quercetin augments activator protein-1 and does not reduce nuclear factor-kappa B in the renal cortex of rats with established chronic glomerular disease. Nephron. 2002;90:313-9.
  • Granado-Serrano AB, Martin MA, Bravo L, Goya L, Ramos S. Quercetin attenuates TNF-induced inflammation in hepatic cells by inhibiting the NF-kappaB pathway. Nutr Cancer. 2012;64:588-98.
  • Jeon H, Kim H, Choi D, Kim D, Park SY, Kim YJ et al. Quercetin activates an angiogenic pathway, hypoxia inducible factor (HIF)-1-vascular endothelial growth factor, by inhibiting HIF-prolyl hydroxylase: a structural analysis of quercetin for inhibiting HIF-prolyl hydroxylase. Mol Pharmacol. 2007;71:1676-84.
  • Hellwig-Burgel T, Rutkowski K, Metzen E, Fandrey J, Jelkmann W. Interleukin-1beta and tumor necrosis factor-alpha stimulate DNA binding of hypoxia-inducible factor-1. Blood. 1999;94:1561-7.
  • Belaiba RS, Bonello S, Zahringer C, Schmidt S, Hess J, Kietzmann T et al. Hypoxia up-regulates hypoxia-inducible factor-1alpha transcription by involving phosphatidylinositol 3-kinase and nuclear factor kappaB in pulmonary artery smooth muscle cells. Mol Biol Cell. 2007;18:4691-7.
  • Ke S, Rabson AB, Germino JF, Gallo MA, Tian Y. Mechanism of suppression of cytochrome P-450 1A1 expression by tumor necrosis factor-alpha and lipopolysaccharide. J Biol Chem. 2001;276:39638-44.
  • Gires O, Kieu C, Fix P, Schmitt B, Munz M, Wollenberg B et al. Tumor necrosis factor alpha negatively regulates the expression of the carcinoma-associated antigen epithelial cell adhesion molecule. Cancer. 2001;92:620-8.
  • Krajewski J, Batmunkh C, Jelkmann W, Hellwig-Burgel T. Interleukin-1beta inhibits the hypoxic inducibility of the erythropoietin enhancer by suppressing hepatocyte nuclear factor-4alpha. Cell Mol Life Sci. 2007;64:989-98.
  • Bunn HF, Gu J, Huang LE, Park JW, Zhu H. Erythropoietin: a model system for studying oxygen-dependent gene regulation. J Exp Biol. 1998;201:1197-201.
  • McGettrick AF, O'Neill LAJ. The Role of HIF in immunity and inflammation. Cell Metab. 2020;32:524-36.
  • Yi YS. Regulatory roles of flavonoids on inflammasome activation during inflammatory responses. Mol Nutr Food Res. 2018;62:e1800147.
  • Malik A, Kanneganti TD. Inflammasome activation and assembly at a glance. J Cell Sci. 2017;130:3955-63.
  • Zhang W, Jia L, Zhao B, Xiong Y, Wang YN, Liang J et al. Quercetin reverses TNF alpha induced osteogenic damage to human periodontal ligament stem cells by suppressing the NFkappaB/NLRP3 inflammasome pathway. Int J Mol Med. 2021;47:39.
  • Choe JY, Kim SK. Quercetin and ascorbic acid suppress fructose-induced NLRP3 inflammasome activation by blocking intracellular shuttling of txnip in human macrophage cell lines. Inflammation. 2017;40:980-94.
  • Xue Y, Du M, Zhu MJ. Quercetin suppresses NLRP3 inflammasome activation in epithelial cells triggered by Escherichia coli O157:H7. Free Radic Biol Med. 2017;10:760-9.
  • Reyes-Farias M, Carrasco-Pozo C. The anti-cancer effect of quercetin: molecular implications in cancer metabolism. Int J Mol Sci. 2019;20:3177.
  • Jana N, Bretislav G, Pavel S, Pavla U. Potential of the flavonoid quercetin to prevent and treat cancer - current status of research. Klin Onkol. 2018;31:184-190.
  • Kabadere S, Oztopcu-Vatan P, Uyar R, Durmaz R. Quercetin both partially attenuates hydrogen peroxide-induced toxicity and decreases viability of rat glial cells. Acta Biol Hung. 2011;62:221-7.
  • Oztopcu-Vatan P, Kabadere S, Uyar R. The effect of pretreatment or combined treatment of quercetin on menadione toxicity in rat primary mixed glial cells in vitro. Cytotechnology. 2009;61:11-6
There are 33 citations in total.

Details

Primary Language English
Subjects Clinical Sciences
Journal Section Research
Authors

Mete Ozkurt 0000-0003-4000-2345

Burcu Akıncı 0000-0002-4316-1950

Selda Kabadere Deliorman 0000-0002-9589-0063

Rumeysa Özyurt 0000-0002-9887-2645

Abdullah Karadağ 0000-0003-3620-9589

Nilüfer Erkasap 0000-0002-2742-3519

Project Number 2018-2004
Publication Date March 31, 2023
Acceptance Date March 15, 2023
Published in Issue Year 2023 Volume: 48 Issue: 1

Cite

MLA Ozkurt, Mete et al. “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”. Cukurova Medical Journal, vol. 48, no. 1, 2023, pp. 268-77, doi:10.17826/cumj.1174607.