Reoxygenation induces reactive oxygen species production and ferroptosis in renal tubular epithelial&nbsp;cells by activating aryl hydrocarbon receptor

Eleftheriadis,Theodoros; Pissas,Georgios; Filippidis,Georgios; Liakopoulos,Vassilios; Stefanidis,Ioannis

doi:10.3892/mmr.2020.11679

Reoxygenation induces reactive oxygen species production and ferroptosis in renal tubular epithelial cells by activating aryl hydrocarbon receptor

Authors:
- Theodoros Eleftheriadis
- Georgios Pissas
- Georgios Filippidis
- Vassilios Liakopoulos
- Ioannis Stefanidis
View Affiliations

Affiliations: Department of Nephrology, Faculty of Medicine, University of Thessaly, 41110 Larissa, Greece
Published online on: November 10, 2020 https://doi.org/10.3892/mmr.2020.11679
Article Number: 41
Copyright: © Eleftheriadis et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

During the reperfusion phase of ischemia‑reperfusion injury, reactive oxygen species (ROS) production aggravates the course of many diseases, including acute kidney injury. Among the various enzymes implicated in ROS production are the enzymes of the cytochromes P450 superfamily (CYPs). Since arylhydrocarbon receptor (AhR) controls the expression of certain CYPs, the involvement of this pathway was evaluated in reperfusion injury. Because AhR may interact with the nuclear factor erythroid 2‑related factor 2 (Nrf2) and the hypoxia‑inducible factor‑1α (HIF‑1α), whether such an interaction takes place and affects reperfusion injury was also assessed. Proximal renal proximal tubular epithelial cells were subjected to anoxia and subsequent reoxygenation. At the onset of reoxygenation, the AhR inhibitor CH223191, the HIF‑1α activator roxadustat, or the ferroptosis inhibitor α‑tocopherol were used. The activity of AhR, Nrf2, HIF‑1α, and their transcriptional targets were assessed with western blotting. ROS production, lipid peroxidation and cell death were measured with colorimetric assays or cell imaging. Reoxygenation induced ROS production, lipid peroxidation and cell ferroptosis, whereas CH223191 prevented all. Roxadustat did not affect the above parameters. Reoxygenation activated AhR and increased CYP1A1, while CH223191 prevented both. Reoxygenation with or without CH223191 did not alter Nrf2 or HIF‑1α activity. Thus, AhR is activated during reoxygenation and induces ROS production, lipid peroxidation and ferroptotic cell death. These detrimental effects may be mediated by AhR‑induced CYP overexpression, while the Nrf2 or the HIF‑1α pathways remain unaffected. Accordingly, the AhR pathway may represent a promising therapeutic target for the prevention of reperfusion injury.

View Figures

View References

1	Neri M, Riezzo I, Pascale N, Pomara C and Turillazzi E: Ischemia/reperfusion injury following acute myocardial infarction: a critical issue for clinicians and forensic pathologists. Mediators Inflamm. 2017:70183932017. View Article : Google Scholar : PubMed/NCBI
2	Bakthavachalam P and Shanmugam PST: Mitochondrial dysfunction - silent killer in cerebral ischemia. J Neurol Sci. 375:417–423. 2017. View Article : Google Scholar : PubMed/NCBI
3	Tsukamoto T, Chanthaphavong RS and Pape HC: Current theories on the pathophysiology of multiple organ failure after trauma. Injury. 41:21–26. 2010. View Article : Google Scholar : PubMed/NCBI
4	Bonventre JV and Yang L: Cellular pathophysiology of ischemic acute kidney injury. J Clin Invest. 121:4210–4221. 2011. View Article : Google Scholar : PubMed/NCBI
5	Eleftheriadis T, Pissas G, Antoniadi G, Liakopoulos V and Stefanidis I: Cell death patterns due to warm ischemia or reperfusion in renal tubular epithelial cells originating from human, mouse, or the native hibernator hamster. Biology (Basel). 7:482018.
6	Eleftheriadis T, Pissas G, Liakopoulos V and Stefanidis I: Factors that may protect the native hibernator syrian hamster renal tubular epithelial cells from ferroptosis due to warm anoxia-reoxygenation. Biology (Basel). 8:222019.
7	Linkermann A, Skouta R, Himmerkus N, Mulay SR, Dewitz C, De Zen F, Prokai A, Zuchtriegel G, Krombach F, Welz PS, et al: Synchronized renal tubular cell death involves ferroptosis. Proc Natl Acad Sci USA. 111:16836–16841. 2014. View Article : Google Scholar : PubMed/NCBI
8	Galluzzi L, Bravo-San Pedro JM, Vitale I, Aaronson SA, Abrams JM, Adam D, Alnemri ES, Altucci L, Andrews D, Annicchiarico-Petruzzelli M, et al: Essential versus accessory aspects of cell death: Recommendations of the NCCD 2015. Cell Death Differ. 22:58–73. 2015. View Article : Google Scholar : PubMed/NCBI
9	Hrycay EG and Bandiera SM: Monooxygenase, peroxidase and peroxygenase properties and reaction mechanisms of cytochrome P450 enzymes. 851:1–61. 2015.PubMed/NCBI
10	Lewis DFV: Oxidative stress: The role of cytochromes P450 in oxygen activation. J Chem Technol Biotechnol. 77:1095–1100. 2002. View Article : Google Scholar
11	Zeng Q, Han Y, Bao Y, Li W, Li X, Shen X, Wang X, Yao F, O'Rourke ST and Sun C: 20-HETE increases NADPH oxidase-derived ROS production and stimulates the L-type Ca2+ channel via a PKC-dependent mechanism in cardiomyocytes. Am J Physiol Heart Circ Physiol. 299:H1109–H1117. 2010. View Article : Google Scholar : PubMed/NCBI
12	Han Y, Zhao H, Tang H, Li X, Tan J, Zeng Q and Sun C: 20-Hydroxyeicosatetraenoic acid mediates isolated heart ischemia/reperfusion injury by increasing NADPH oxidase-derived reactive oxygen species production. Circ J. 77:1807–1816. 2013. View Article : Google Scholar : PubMed/NCBI
13	Granville DJ, Tashakkor B, Takeuchi C, Gustafsson AB, Huang C, Sayen MR, Wentworth P Jr, Yeager M and Gottlieb RA: Reduction of ischemia and reperfusion-induced myocardial damage by cytochrome P450 inhibitors. Proc Natl Acad Sci USA. 101:1321–1326. 2004. View Article : Google Scholar : PubMed/NCBI
14	Ishihara Y, Sekine M, Nakazawa M and Shimamoto N: Suppression of myocardial ischemia-reperfusion injury by inhibitors of cytochrome P450 in rats. Eur J Pharmacol. 611:64–71. 2009. View Article : Google Scholar : PubMed/NCBI
15	Ishihara Y, Sekine M, Hamaguchi A, Kobayashi Y, Harayama T, Nakazawa M and Shimamoto N: Effects of sulfaphenazole derivatives on cardiac ischemia-reperfusion injury: Association of cytochrome P450 activity and infarct size. J Pharmacol Sci. 113:335–342. 2010. View Article : Google Scholar : PubMed/NCBI
16	Shaik IH and Mehvar R: Cytochrome P450 induction by phenobarbital exacerbates warm hepatic ischemia-reperfusion injury in rat livers. Free Radic Res. 44:441–453. 2010. View Article : Google Scholar : PubMed/NCBI
17	Shaik IH and Mehvar R: Effects of cytochrome p450 inhibition by cimetidine on the warm hepatic ischemia-reperfusion injury in rats. J Surg Res. 159:680–688. 2010. View Article : Google Scholar : PubMed/NCBI
18	Nebert DW, Dalton TP, Okey AB and Gonzalez FJ: Role of aryl hydrocarbon receptor-mediated induction of the CYP1 enzymes in environmental toxicity and cancer. J Biol Chem. 279:23847–23850. 2004. View Article : Google Scholar : PubMed/NCBI
19	Lindsey S and Papoutsakis ET: The evolving role of the aryl hydrocarbon receptor (AHR) in the normophysiology of hematopoiesis. Stem Cell Rev Rep. 8:1223–1235. 2012. View Article : Google Scholar : PubMed/NCBI
20	Pollenz RS: The mechanism of AH receptor protein down-regulation (degradation) and its impact on AH receptor-mediated gene regulation. Chem Biol Interact. 141:41–61. 2002. View Article : Google Scholar : PubMed/NCBI
21	Ma Q and Baldwin KT: 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced degradation of aryl hydrocarbon receptor (AhR) by the ubiquitin-proteasome pathway. Role of the transcription activaton and DNA binding of AhR. J Biol Chem. 275:8432–8438. 2000. View Article : Google Scholar : PubMed/NCBI
22	Forrester AR, Elias MS, Woodward EL, Graham M, Williams FM and Reynolds NJ: Induction of a chloracne phenotype in an epidermal equivalent model by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is dependent on aryl hydrocarbon receptor activation and is not reproduced by aryl hydrocarbon receptor knock down. J Dermatol Sci. 73:10–22. 2014. View Article : Google Scholar : PubMed/NCBI
23	Yang SC, Wu CH, Tu YK, Huang SY and Chou PC: Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin increases the activation of aryl hydrocarbon receptor and is associated with the aggressiveness of osteosarcoma MG-63 osteoblast-like cells. Oncol Lett. 16:3849–3857. 2018.PubMed/NCBI
24	Moretti S, Nucci N, Menicali E, Morelli S, Bini V, Colella R, Mandarano M, Sidoni A and Puxeddu E: The Aryl Hydrocarbon Receptor Is Expressed in Thyroid Carcinoma and Appears to Mediate Epithelial-Mesenchymal-Transition. Cancers (Basel). 12:1452020. View Article : Google Scholar
25	Couroucli XI, Liang YW, Jiang W, Barrios R and Moorthy B: Attenuation of oxygen-induced abnormal lung maturation in rats by retinoic acid: Possible role of cytochrome P4501A enzymes. J Pharmacol Exp Ther. 317:946–954. 2006. View Article : Google Scholar : PubMed/NCBI
26	Kopf PG, Scott JA, Agbor LN, Boberg JR, Elased KM, Huwe JK and Walker MK: Cytochrome P4501A1 is required for vascular dysfunction and hypertension induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Toxicol Sci. 117:537–546. 2010. View Article : Google Scholar : PubMed/NCBI
27	Kopf PG and Walker MK: 2,3,7,8-tetrachlorodibenzo-p-dioxin increases reactive oxygen species production in human endothelial cells via induction of cytochrome P4501A1. Toxicol Appl Pharmacol. 245:91–99. 2010. View Article : Google Scholar : PubMed/NCBI
28	Lee HJ, Pyo MC, Shin HS, Ryu D and Lee K-W: Renal toxicity through AhR, PXR, and Nrf2 signaling pathway activation of ochratoxin A-induced oxidative stress in kidney cells. Food Chem Toxicol. 122:59–68. 2018. View Article : Google Scholar : PubMed/NCBI
29	Pei J, Juni R, Harakalova M, Duncker DJ, Asselbergs FW, Koolwijk P, Hinsbergh VV, Verhaar MC, Mokry M and Cheng C: Indoxyl sulfate stimulates angiogenesis by regulating reactive oxygen species production via CYP1B1. Toxins (Basel). 11:4542019. View Article : Google Scholar
30	Furue M, Takahara M, Nakahara T and Uchi H: Role of AhR/ARNT system in skin homeostasis. Arch Dermatol Res. 306:769–779. 2014. View Article : Google Scholar : PubMed/NCBI
31	Yagishita Y, Uruno A and Yamamoto M: NRF2-mediated gene regulation and glucose homeostasis. Molecular nutrition and diabetes. Elsevier Inc.; pp. 331–348. 2016, View Article : Google Scholar
32	Dietrich C: Antioxidant functions of the aryl hydrocarbon receptor. Stem Cells Int. 2016:79434952016. View Article : Google Scholar : PubMed/NCBI
33	Ma Q: Role of nrf2 in oxidative stress and toxicity. Annu Rev Pharmacol Toxicol. 53:401–426. 2013. View Article : Google Scholar : PubMed/NCBI
34	Corsello T, Komaravelli N and Casola A: Role of hydrogen sulfide in NRF2- and sirtuin-dependent maintenance of cellular redox balance. Antioxidants. 7:1292018. View Article : Google Scholar
35	Cuartero MI, Ballesteros I, de la Parra J, Harkin AL, Abautret-Daly A, Sherwin E, Fernández-Salguero P, Corbí AL, Lizasoain I and Moro MA: L-kynurenine/aryl hydrocarbon receptor pathway mediates brain damage after experimental stroke. Circulation. 130:2040–2051. 2014. View Article : Google Scholar : PubMed/NCBI
36	Kaluz S, Kaluzová M and Stanbridge EJ: Regulation of gene expression by hypoxia: Integration of the HIF-transduced hypoxic signal at the hypoxia-responsive element. Clin Chim Acta. 395:6–13. 2008. View Article : Google Scholar : PubMed/NCBI
37	Myllyharju J: Prolyl 4-hydroxylases, master regulators of the hypoxia response. Acta Physiol (Oxf). 208:148–165. 2013. View Article : Google Scholar : PubMed/NCBI
38	Vorrink SU, Sarsour EH, Olivier AK, Robertson LW, Goswami PC and Domann FE: PCB 126 perturbs hypoxia-induced HIF-1α activity and glucose consumption in human HepG2 cells. Exp Toxicol Pathol. 66:377–382. 2014. View Article : Google Scholar : PubMed/NCBI
39	Vorrink SU, Severson PL, Kulak MV, Futscher BW and Domann FE: Hypoxia perturbs aryl hydrocarbon receptor signaling and CYP1A1 expression induced by PCB 126 in human skin and liver-derived cell lines. Toxicol Appl Pharmacol. 274:408–416. 2014. View Article : Google Scholar : PubMed/NCBI
40	Eleftheriadis T, Pissas G, Antoniadi G, Liakopoulos V and Stefanidis I: Kynurenine, by activating aryl hydrocarbon receptor, decreases erythropoietin and increases hepcidin production in HepG2 cells: A new mechanism for anemia of inflammation. Exp Hematol. 44:60–7.e1. 2016. View Article : Google Scholar : PubMed/NCBI
41	Kim SH, Henry EC, Kim DK, Kim YH, Shin KJ, Han MS, Lee TG, Kang JK, Gasiewicz TA, Ryu SH, et al: Novel compound 2-methyl-2H-pyrazole-3-carboxylic acid (2-methyl-4-o-tolylazo-phenyl)-amide (CH-223191) prevents 2,3,7,8-TCDD-induced toxicity by antagonizing the aryl hydrocarbon receptor. Mol Pharmacol. 69:1871–1878. 2006. View Article : Google Scholar : PubMed/NCBI
42	Dhillon S: Roxadustat: First Global Approval. Drugs. 79:563–572. 2019. View Article : Google Scholar : PubMed/NCBI
43	Stockwell BR, Friedmann Angeli JP, Bayir H, Bush AI, Conrad M, Dixon SJ, Fulda S, Gascón S, Hatzios SK, Kagan VE, et al: Ferroptosis: A regulated cell death nexus linking metabolism, redox biology, and disease. Cell. 171:273–285. 2017. View Article : Google Scholar : PubMed/NCBI
44	Rzemieniec J, Wnuk A, Lasoń W, Bilecki W and Kajta M: The neuroprotective action of 3,3′-diindolylmethane against ischemia involves an inhibition of apoptosis and autophagy that depends on HDAC and AhR/CYP1A1 but not ERα/CYP19A1 signaling. Apoptosis. 24:435–452. 2019. View Article : Google Scholar : PubMed/NCBI
45	Stejskalova L, Dvorak Z and Pavek P: Endogenous and exogenous ligands of aryl hydrocarbon receptor: Current state of art. Curr Drug Metab. 12:198–212. 2011. View Article : Google Scholar : PubMed/NCBI
46	Wu PY, Chuang PY, Chang GD, Chan YY, Tsai TC, Wang BJ, Lin KH, Hsu WM, Liao YF and Lee H: Novel endogenous ligands of aryl hydrocarbon receptor mediate neural development and differentiation of neuroblastoma. ACS Chem Neurosci. 10:4031–4042. 2019. View Article : Google Scholar : PubMed/NCBI
47	Fadeel B and Orrenius S: Apoptosis: A basic biological phenomenon with wide-ranging implications in human disease. J Intern Med. 258:479–517. 2005. View Article : Google Scholar : PubMed/NCBI
48	Huang B, Bao J, Cao YR, Gao HF and Jin Y: Cytochrome P450 1A1 (CYP1A1) catalyzes lipid peroxidation of oleic acid-induced HepG2 cells. Biochemistry (Mosc). 83:595–602. 2018. View Article : Google Scholar : PubMed/NCBI
49	Mohib K, Wang S, Guan Q, Mellor AL, Sun H, Du C and Jevnikar AM: Indoleamine 2,3-dioxygenase expression promotes renal ischemia-reperfusion injury. Am J Physiol Renal Physiol. 295:F226–F234. 2008. View Article : Google Scholar : PubMed/NCBI
50	Fang YZ, Yang S and Wu G: Free radicals, antioxidants, and nutrition. Nutrition. 18:872–879. 2002. View Article : Google Scholar : PubMed/NCBI
51	Eleftheriadis T, Pissas G, Nikolaou E, Liakopoulos V and Stefanidis I: The H2S-Nrf2-antioxidant proteins axis protects renal tubular epithelial cells of the native hibernator syrian hamster from reoxygenation-induced cell death. Biology (Basel). 8:742019.