The Protective Effects of Quercetin on Ferritinophagy in Alcoholic-Induced Liver Iron Overload

Abstract

Objective

Iron overload resulting from chronic alcohol consumption may aggravate liver damage, and the potential mechanisms involving ferritinophagic flux and the role of naturally occurring quercetin in alcohol-induced liver disease remain unclear.

Methods

Adult male C57BL/6J mice were iso-calorically pair-fed with ethanol-containing Lieber De Carli liquid diets according to a chronic-plus-binge ethanol feeding protocol with either quercetin (100 mg/kg.bw) or iron-rich/limited treatment for 12 weeks, and liver damage, as well as the underlying mechanisms of lysosome-dependent ferritinophagy, was explored, following the study of ethanol-incubated HepG2 cells with specific pharmacological reagents or gene regulation in vitro.

Results

Chronic-plus-binge ethanol feeding led to an increase in the hepatosomatic ratio, hepatic lipid accumulation and triglyceride (TG) content of the mice and induced the release of alanine aminotransferase (ALT), aspartate transaminase (AST), and serum TG levels, which were normalized partially by quercetin treatment or iron limitation but worsened by iron supplementation. Similar findings were observed in vitro. Moreover, quercetin intervention alleviated iron deposition, inhibited the upregulation of p62 and downregulation of nuclear receptor coactivator 4 (NCOA4) and microtubule-associated protein 1 light chain 3 (LC3)-II, and blocked the colocalization of NCOA4 and ferritin heavy chain and the nuclear translocation of forkhead box protein O1 (FOXO1) induced by ethanol. These effects were also observed when the cells were subjected to iron limitation but were abolished by iron supplementation, NCOA4 transfection, or AS1842856, a FOXO1 inhibitor.

Conclusion

Quercetin ameliorates secondary iron overload and subsequent liver damage caused by alcohol abuse by maintaining ferritinophagic flux and lysosome function via the FOXO1-TFEB NCOA4 signaling pathway.

Keywords

Alcoholic liver disease / Iron overload / Quercetin / Ferritinophagy / Nuclear receptor coactivator 4 / Transcription factor EB

References

[1] Peacock A, Leung J, Larney S, et al.. Global statistics on alcohol, tobacco and illicit drug use: 2017 status report. Addiction., 2018, 113(10): 1905-1926

[2] GBD 2016 Alcohol Collaborators. Alcohol use and burden for 195 countries and territories, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet. 2018;392(10152): 1015–1035.

[3] Singal AK, Mathurin P. Diagnosis and Treatment of Alcohol-Associated Liver Disease. JAMA., 2021, 326(2): 165-176

[4] Li LX, Guo FF, Liu H, et al.. Iron overload in alcoholic liver disease: underlying mechanisms, detrimental effects, and potential therapeutic targets. Cell Mol Life Sci., 2022, 79(4): 201

[5] Axley PD, Richardson CT, Singal AK. Epidemiology of Alcohol Consumption and Societal Burden of Alcoholism and Alcoholic Liver Disease. Clin Liver Dis., 2019, 23(139-50

[6] Duan Y, Chu H, Brandl K, et al.. CRIg on liver macrophages clears pathobionts and protects against alcoholic liver disease. Nat Commun., 2021, 12(1): 7172

[7] Wang CY, Babitt JL. Liver iron sensing and body iron homeostasis. Blood., 2019, 133(118-29

[8] Magdaleno F, Blajszczak C, Nieto N. Key Events Participating in the Pathogenesis of Alcoholic Liver Disease. Biomolecules., 2017, 7(1): 9

[9] Radicheva MP, Andonova AN, Milcheva HT, et al.. Serum Markers of Iron Metabolism in Chronic Liver Diseases. Open Access Maced J Med Sci., 2018, 6(61010-1016

[10] Mancias JD, Wang X, Gygi SP, et al.. Quantitative proteomics identifies NCOA4 as the cargo receptor mediating ferritinophagy. Nature., 2014, 509(7498): 105-109

[11] Mancias JD, Pontano Vaites L, Nissim S, et al.. Ferritinophagy via NCOA4 is required for erythropoiesis and is regulated by iron dependent HERC2-mediated proteolysis. Elife., 2015, 5(4 e10308

[12] Ryu MS, Duck KA, Philpott CC. Ferritin iron regulators, PCBP1 and NCOA4, respond to cellular iron status in developing red cells. Blood Cells Mol Dis., 2018, 69: 75-81

[13] Sun Y, Bao Q, Xuan B, et al.. Human Cytomegalovirus Protein pUL38 Prevents Premature Cell Death by Binding to Ubiquitin-Specific Protease 24 and Regulating Iron Metabolism. J Virol., 2018, 92(13): e00191-e218

[14] Yang H, Sun WS, Bi T, et al.. The PTBP1-NCOA4 axis promotes ferroptosis in liver cancer cells. Oncol Rep., 2023, 49(2): 45

[15] Di Petrillo A, Orrù G, Fais A, et al.. Quercetin and its derivates as antiviral potentials: A comprehensive review. Phytother Res., 2021, 36(1266-278

[16] Yu X, Xu YY, Zhang SS, et al. Quercetin Attenuates Chronic Ethanol-Induced Hepatic Mitochondrial Damage through Enhanced Mitophagy. Nutrients. 2016;5;8(1):27.

[17] Zeng H, Guo X, Zhou F, et al.. Quercetin alleviates ethanol-induced liver steatosis associated with improvement of lipophagy. Food Chem Toxicol., 2019, 125: 21-28

[18] Gryzik M, Srivastava A, Longhi G, et al. Expression and characterization of the ferritin binding domain of Nuclear Receptor Coactivator-4 (NCOA4). Bioch Biophysa Acta Gen Subj. 2017;1861(11 Pt A):2710–2716.

[19] Ohshima T, Yamamoto H, Sakamaki Y, et al. NCOA4 drives ferritin phase separation to facilitate macroferritinophagy and microferritinophagy. J Cell Biol. 2022;3;221(10):e202203102.

[20] He Y, Cao X, Guo P, et al.. Quercetin induces autophagy via FOXO1-dependent pathways and autophagy suppression enhances quercetin-induced apoptosis in PASMCs in hypoxia. Free Radic Biol Med., 2017, 103: 165-176

[21] Wang J, Zhou J, Mo WC, et al.. Accumulation of simulated pathological level of formaldehyde decreases cell viability and adhesive morphology in neuronal cells. Prog Biochem Biophys., 2017, 44(7): 601-614

[22] Liu L, Tao Z, Zheng LD, et al.. FoxO1 interacts with transcription factor EB and differentially regulates mitochondrial uncoupling proteins via autophagy in adipocytes. Cell Death Discov., 2016, 3(2): 16066

[23] Asrani SK, Devarbhavi H, Eaton J, et al.. Burden of liver diseases in the world. J Hepatol., 2019, 70(1151-171

[24] Ramirez T, Li YM, Yin S, et al.. Aging aggravates alcoholic liver injury and fibrosis in mice by downregulating sirtuin 1 expression. J Hepatol., 2017, 66(3): 601-609

[25] Lamas-Paz A, Hao F, Nelson LJ, et al. Alcoholic liver disease: Utility of animal models. World J Gastroenterol. 2018;7;24(45):5063–5075.

[26] Simpson RF, Hermon C, Liu B, et al.. Alcohol drinking patterns and liver cirrhosis risk: analysis of the prospective UK Million Women Study. Lancet Public Health., 2019, 4(1): e41-e48

[27] Li S, Cao H, Shen D, et al. Quercetin protects against ox‑LDL‑induced injury via regulation of ABCAl, LXR‑α and PCSK9 in RAW264. 7 macrophages. Mol Med Rep. 2018;18(1):799–806.

[28] Roslan J, Giribabu N, Karim K, et al.. Quercetin ameliorates oxidative stress, inflammation and apoptosis in the heart of streptozotocin-nicotinamide-induced adult male diabetic rats. Biomed Pharmacother., 2017, 86: 570-582

[29] Guazelli CFS, Staurengo-Ferrari L, Zarpelon AC, et al.. Quercetin attenuates zymosan-induced arthritis in mice. Biomed Pharmacother., 2018, 102: 175-184

[30] Liu S, Tian L, Chai G, et al. Targeting heme oxygenase-1 by quercetin ameliorates alcohol-induced acute liver injury via inhibiting NLRP3 inflammasome activation. Food Funct. 2018;15;9(8):4184–4193.

[31] Xiao L, Luo G, Tang Y, et al.. Quercetin and iron metabolism: What we know and what we need to know. Food Chem Toxicol., 2018, 114: 190-203

[32] El-Sheikh AA, Ameen SH, Abdel-Fatah SS. Ameliorating Iron Overload in Intestinal Tissue of Adult Male Rats: Quercetin vs Deferoxamine. J Toxicol., 2018, 2018(21): 8023840

[33] Arosio P, Elia L, Poli M. Ferritin, cellular iron storage and regulation. IUBMB Life., 2017, 69(6414-422

[34] Gupta P, Singh P, Pandey HS, et al.. Phosphoinositide-3-kinase inhibition elevates ferritin level resulting depletion of labile iron pool and blocking of glioma cell proliferation. Biochim Biophys Acta Gen Subj., 2019, 1863(3): 547-564

[35] Melle C, Ernst G, Scheibner O, et al.. Identification of Specific Protein Markers in Microdissected Hepatocellular Carcinoma. J Proteome Res., 2006, 6(1): 306-315

[36] Dowdle WE, Nyfeler B, Nagel J, et al.. Selective VPS34 inhibitor blocks autophagy and uncovers a role for NCOA4 in ferritin degradation and iron homeostasis in vivo. Nat Cell Biol., 2014, 16(11): 1069-1079

[37] Bellelli R, Federico G, Matte’ A, et al. NCOA4 Deficiency Impairs Systemic Iron Homeostasis. Cell Rep. 2016;14(3):411–421.

[38] Yao Z, Gu Y, Zhang Q, et al.. Estimated daily quercetin intake and association with the prevalence of type 2 diabetes mellitus in Chinese adults. Eur J Nutr., 2018, 58(2): 819-830

[39] Das SK, Desaulniers J, Dyck JRB, et al.. Resveratrol mediates therapeutic hepatic effects in acquired and genetic murine models of iron-overload. Liver Int., 2015, 36(2): 246-257

[40] Masaldan S, Clatworthy SA, Gamell C, et al.. Iron accumulation in senescent cells is coupled with impaired ferritinophagy and inhibition of ferroptosis. Redox biol., 2018, 14: 100-115

[41] Huang Y, Chen Y, Shaw AM, et al.. Enhancing TFEB-Mediated Cellular Degradation Pathways by the mTORC1 Inhibitor Quercetin. Oxid Med Cell Longev., 2018, 2018(285073420

Funding

National Natural Science Foundation of China(82273628)

RIGHTS & PERMISSIONS

The Author(s), under exclusive licence to the Huazhong University of Science and Technology