Abstract
Iron is an essential element for the normal functioning of living organisms, but excessive iron deposition can lead to organ damage. This study aims to investigate the interaction between the endoplasmic reticulum stress signaling pathway and the PI3K/AKT/mTOR signaling pathway in liver injury induced by iron overload in chicks. Rspectively, 150 one-day-old broilers were divided into three groups and supplemented with 50 (C), 500 (E1), and 1000 (E2) mg ferrous sulfate monohydrate/kg in the basal diet. Samples were taken after continuous feeding for 14 days. The results showed that iron overload could upregulate the levels of ALT and AST. Histopathological examination revealed bleeding in the central vein of the liver accompanied by inflammatory cell infiltration. Hoechst staining showed that the iron overload group showed significant bright blue fluorescence, and ultrastructural observations showed chromatin condensation as well as mitochondrial swelling and cristae disorganization in the iron overload group. RT-qPCR and Western blot results showed that iron overload upregulated the expression of Bax, Caspase-3, Caspase-9, GRP78, GRP94, P-PERK, ATF4, eIF2α, IRE1, and ATF6, while downregulating the expression of Bcl-2 and the PI3K/AKT/mTOR pathway. XBP-1 splicing experiment showed significant splicing of XBP-1 gene after iron overload. PCA and correlation analysis suggested a potential association between endoplasmic reticulum stress, the PI3K/AKT/mTOR signaling pathway, and liver injury in chicks. In summary, iron overload can induce cell apoptosis and liver injury by affecting endoplasmic reticulum stress and the PI3K/AKT/mTOR signaling pathway.
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Abbina S, Abbasi U, Gill A, Leitch H, Kizhakkedathu JN (2022) Active transport nanochelators for the reduction of liver iron burden in iron overload. J Control Rel 350:857–869. https://doi.org/10.1016/j.jconrel.2022.08.056
Almanza A, Carlesso A, Chintha C, Creedican S, Doultsinos D, Leuzzi B, Luís A, McCarthy N, Montibeller L, More S, Papaioannou A, Püschel F, Sassano ML, Skoko J, Agostinis P, de Belleroche J, Eriksson LA, Fulda S, Gorman AM, Healy S, Kozlov A, Muñoz-Pinedo C, Rehm M, Chevet E, Samali A (2019) Endoplasmic reticulum stress signalling – from basic mechanisms to clinical applications. FEBS J 286(2):241–278. https://doi.org/10.1111/febs.14608
Ashkenazi A, Fairbrother WJ, Leverson JD, Souers AJ (2017) From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors. Nat Rev Drug Discovery 16(4):273–284. https://doi.org/10.1038/nrd.2016.253
Cheng X, Ferrell JE Jr (2018) Apoptosis propagates through the cytoplasm as trigger waves. Science (New York, N.Y.) 361(6402):607–612. https://doi.org/10.1126/science.aah4065
Cherubini A, Zito E (2022) ER stress as a trigger of UPR and ER-phagy in cancer growth and spread. Front Oncol 12:997235. https://doi.org/10.3389/fonc.2022.997235
Cudd SK, Garner MM, Cartoceti AN, LaDouceur EEB (2022) Hepatic lesions associated with iron accumulation in captive kori bustards (Ardeotis kori). Vet Pathol 59(1):164–168. https://doi.org/10.1177/03009858211035386
Ding H, Zhang Q, Yu X, Chen L, Wang Z, Feng J (2021) Lipidomics reveals perturbations in the liver lipid profile of iron-overloaded mice. Metallomics Integ Biometal Sci 13(10). https://doi.org/10.1093/mtomcs/mfab057
Fang Z, Yushanjiang F, Wang G, Zheng X, Jiang X (2023) Germacrone mitigates cardiac remodeling by regulating PI3K/AKT-mediated oxidative stress, inflammation, and apoptosis. Int Immunopharmacol 124(Pt A):110876. https://doi.org/10.1016/j.intimp.2023.110876
Feng L, Liao H, Liu J, Xu C, Zhong K, Zhu H, Guo S, Guo Y, Han L, Li H, Wang Y (2021) Inhibition of PI3K/Akt/mTOR pathway by ammonium chloride induced apoptosis and autophagy in MAC-T cell. Res Vet Sci 136:622–630. https://doi.org/10.1016/j.rvsc.2021.01.020
Gao H, He C, Hua R, Guo Y, Wang B, Liang C, Gao L, Shang H, Xu JD (2022) Endoplasmic reticulum stress of gut enterocyte and intestinal diseases. Front Mol Biosci 9:817392. https://doi.org/10.3389/fmolb.2022.817392
Glimcher LH, Lee AH, Iwakoshi NN (2020) XBP-1 and the unfolded protein response (UPR). Nat Immunol 21(9):963–965. https://doi.org/10.1038/s41590-020-0708-3
James AW, Bahader GA, Albassan M, Shah ZA (2023) The ER chaperone, BIP protects Microglia from ER stress-mediated Apoptosis in Hyperglycemia. Neurochem Int 169:105573. https://doi.org/10.1016/j.neuint.2023.105573
Jatobá CA, de Rezende AA, de Paiva RSJ, de Almeida CMM, das Graças Almeida M, Freire-Neto F, da Rocha LR, da Medeiros AC, Brandão-Neto J, de Carvalho Formiga MC, de Azevedo IM, de Oliveira Ramos AM (2008) Liver iron overload induced by tamoxifen in diabetic and non-diabetic female Wistar rats. Biometals : An Int J Role Metal Ions Biol Biochem Med 21(2):171–178. https://doi.org/10.1007/s10534-007-9105-8
Ji J, Jing A, Ding Y, Ma X, Qian Q, Geng T, Cheng W, Zhang M, Sun Q, Ma S, Wang X, Yuan Q, Xu M, Qin J, Ma L, Yang J, He J, Du Q, Xia M, Xu Y, Chen Z, Zhu L, Liu W, Liu S, Liu B (2024) FBXO5-mediated RNF183 degradation prevents endoplasmic reticulum stress-induced apoptosis and promotes colon cancer progression. Cell Death Dis 15(1):33. https://doi.org/10.1038/s41419-024-06421-2
Jiang Y, Huang Z, Li X, Zhou L, Zhu X, Chen F, Shi Y (2023) Inhibition of SK2 and ER stress ameliorated inflammation and apoptosis in liver ischemia-reperfusion injury. Int Liver Transplant Soc 29(10):1050–1062. https://doi.org/10.1097/lvt.0000000000000210
Lee D, Lee SH, Noh I, Oh E, Ryu H, Ha J, Jeong S, Yoo J, Jeon TJ, Yun CO, Kim YC (2019) A Helical polypeptide-based potassium ionophore induces endoplasmic reticulum stress-mediated apoptosis by perturbing ion homeostasis. Adv Sci (Weinheim, Baden-Wurttemberg, Germany) 6(14):1801995. https://doi.org/10.1002/advs.201801995
Li L, Miao X, Ni R, Miao X, Wang L, Gu X, Yan L, Tang Q, Zhang D (2015) Epithelial-specific ETS-1 (ESE1/ELF3) regulates apoptosis of intestinal epithelial cells in ulcerative colitis via accelerating NF-κB activation. Immunol Res 62(2):198–212. https://doi.org/10.1007/s12026-015-8651-3
Li X, Zhu S, Li Z, Meng YQ, Huang SJ, Yu QY, Li B (2022) Melittin induces ferroptosis and ER stress-CHOP-mediated apoptosis in A549 cells. Free Radical Res 56(5–6):398–410. https://doi.org/10.1080/10715762.2022.2131551
Liu Y, Zhao L, He X, Shen Y, Wang N, Hu S, Xu J, Zhao Q, Zhang Q, Qin L, Zhang Q (2023) Jintiange proteins promote osteogenesis and inhibit apoptosis of osteoblasts by enhancing autophagy via PI3K/AKT and ER stress pathways. J Ethnopharmacol 311:116399. https://doi.org/10.1016/j.jep.2023.116399
Mann J, Reznik E, Santer M, Fongheiser MA, Smith N, Hirschhorn T, Zandkarimi F, Soni RK, Dafré AL, Miranda-Vizuete A, Farina M, Stockwell BR (2023) Ferroptosis inhibition by oleic acid mitigates iron-overload-induced injury. Cell Chem Biol. https://doi.org/10.1016/j.chembiol.2023.10.012
McMillen SA, Nonnecke EB, Lönnerdal B (2022) Trace element interactions, inflammatory signaling, and male sex implicated in reduced growth following excess oral iron supplementation in pre-weanling rats. Nutrients 14(19). https://doi.org/10.3390/nu14193913
Miglioranza Scavuzzi B, Holoshitz J (2022) Endoplasmic reticulum stress, oxidative stress, and rheumatic diseases. Antioxidants (Basel, Switzerland) 11(7). https://doi.org/10.3390/antiox11071306
Nagata S (2018) Apoptosis and clearance of apoptotic cells. Annu Rev Immunol 36:489–517. https://doi.org/10.1146/annurev-immunol-042617-053010
Nemeth E, Ganz T (2021) Hepcidin-Ferroportin interaction controls systemic iron homeostasis. Int J Mol Sci 22(12). https://doi.org/10.3390/ijms22126493
Paliwal S, Jain S, Mudgal P, Verma K, Paliwal S, Sharma S (2023) Mitochondrial transfer restores impaired liver functions by AMPK/ mTOR/PI3K-AKT pathways in metabolic syndrome. Life Sci 332:122116. https://doi.org/10.1016/j.lfs.2023.122116
Rafati RM, Rafati RM, Kazemi S, Moghadamnia AR, Ghaemi AM, Moghadamnia AA (2023) Iron; Benefits or threatens (with emphasis on mechanism and treatment of its poisoning). Hum Exp Toxicol 42:9603271231192360. https://doi.org/10.1177/09603271231192361
Rehman H, Akram M, Kiyani MM, Yaseen T, Ghani A, Saggu JI, Shah SSH, Khalid ZM, Bokhari SAI (2020) Effect of endoxylanase and iron oxide nanoparticles on performance and histopathological features in broilers. Biol Trace El Em Res 193(2):524–535. https://doi.org/10.1007/s12011-019-01737-z
Riegman M, Sagie L, Galed C, Levin T, Steinberg N, Dixon SJ, Wiesner U, Bradbury MS, Niethammer P, Zaritsky A, Overholtzer M (2020) Ferroptosis occurs through an osmotic mechanism and propagates independently of cell rupture. Nat Cell Biol 22(9):1042–1048. https://doi.org/10.1038/s41556-020-0565-1
Rinella ME, Dufour JF, Anstee QM, Goodman Z, Younossi Z, Harrison SA, Loomba R, Sanyal AJ, Bonacci M, Trylesinski A, Natha M, Shringarpure R, Granston T, Venugopal A, Ratziu V (2022) Non-invasive evaluation of response to obeticholic acid in patients with NASH: results from the REGENERATE study. J Hepatol 76(3):536–548. https://doi.org/10.1016/j.jhep.2021.10.029
Seervi M, Rani A, Sharma AK, Santhosh Kumar TR (2018) ROS mediated ER stress induces Bax-Bak dependent and independent apoptosis in response to Thioridazine. Biomed Pharmacother 106:200–209. https://doi.org/10.1016/j.biopha.2018.06.123
Shen X, Niu C, Guo J, Xia M, Xia J, Hu Y, Zheng Y (2018) Stra8 may inhibit apoptosis during mouse spermatogenesis via the AKT signaling pathway. Int J Mol Med 42(5):2819–2830. https://doi.org/10.3892/ijmm.2018.3825
Shen J, Yang Z, Wu X, Yao G, Hou M (2023) Baicalein facilitates gastric cancer cell apoptosis by triggering endoplasmic reticulum stress via repression of the PI3K/AKT pathway. Applied Biol Chem 66(1):10. https://doi.org/10.1186/s13765-022-00759-x
Shojaie L, Iorga A, Dara L (2020) Cell death in liver diseases: a review. Int J Mol Sci 21(24). https://doi.org/10.3390/ijms21249682
Taschetto D, Vieira SL, Angel CR, Stefanello C, Kindlein L, Ebbing MA, Simões CT (2017) Iron requirements of broiler breeder hens. Poult Sci 96(11):3920–3927. https://doi.org/10.3382/ps/pex208
Thorpe LM, Yuzugullu H, Zhao JJ (2015) PI3K in cancer: divergent roles of isoforms, modes of activation and therapeutic targeting. Nat Rev Cancer 15(1):7–24. https://doi.org/10.1038/nrc3860
Tian LY, Smit DJ, Jücker M (2023) The role of PI3K/AKT/mTOR signaling in hepatocellular carcinoma metabolism. Int J Mol Sci 24(3). https://doi.org/10.3390/ijms24032652
Uzhytchak M, Smolková B, Lunova M, Jirsa M, Frtús A, Kubinová Š, Dejneka A, Lunov O (2020) Iron oxide nanoparticle-induced autophagic flux is regulated by interplay between p53-mTOR axis and Bcl-2 signaling in hepatic cells. Cells 9(4). https://doi.org/10.3390/cells9041015
Wang S, Duan J, Liao J, Wang Y, Xiao X, Li L, Liu Y, Gu H, Yang P, Fu D, Du J, Li X Shao M (2022a) LncRNA H19 inhibits ER stress induced apoptosis and improves diabetic cardiomyopathy by regulating PI3K/AKT/mTOR axis. Aging 14(16):6809–6828. https://doi.org/10.18632/aging.204256
Wang Y, Wang M, Liu Y, Tao H, Banerjee S, Srinivasan S, Nemeth E, Czaja MJ, He P (2022b) Integrated regulation of stress responses, autophagy and survival by altered intracellular iron stores. Redox Biol 55:102407. https://doi.org/10.1016/j.redox.2022.102407
Wiseman RL, Mesgarzadeh JS, Hendershot LM (2022) Reshaping endoplasmic reticulum quality control through the unfolded protein response. Mol Cell 82(8):1477–1491. https://doi.org/10.1016/j.molcel.2022.03.025
Xu G, Li X, Zhu Z, Wang H, Bai X (2021) Iron overload induces apoptosis and cytoprotective autophagy regulated by ROS generation in Mc3t3-E1 cells. Biol Trace Elem Res 199(10):3781–3792. https://doi.org/10.1007/s12011-020-02508-x
Yamada N, Karasawa T, Kimura H, Watanabe S, Komada T, Kamata R, Sampilvanjil A, Ito J, Nakagawa K, Kuwata H, Hara S, Mizuta K, Sakuma Y, Sata N, Takahashi M (2020) Ferroptosis driven by radical oxidation of n-6 polyunsaturated fatty acids mediates acetaminophen-induced acute liver failure. Cell Death Dis 11(2):144. https://doi.org/10.1038/s41419-020-2334-2
Yan Y, Liu Y, Yang Y, Ding Y, Sun X (2022) Carnosol suppresses microglia cell inflammation and apoptosis through PI3K/AKT/mTOR signaling pathway. Immunopharmacol Immunotoxicol 44(5):656–662. https://doi.org/10.1080/08923973.2022.2074448
Yan X, Xie Y, Liu H, Huang M, Yang Z, An D, Jiang G (2023) Iron accumulation and lipid peroxidation: implication of ferroptosis in diabetic cardiomyopathy. Diabetol Metab Syndr 15(1):161. https://doi.org/10.1186/s13098-023-01135-5
Yang M, Yao X, Xia F, Xiang S, Tang W, Zhou B (2024) Hugan Qingzhi tablets attenuates endoplasmic reticulum stress in nonalcoholic fatty liver disease rats by regulating PERK and ATF6 pathways. BMC Complement Med Therap 24(1):36. https://doi.org/10.1186/s12906-024-04336-1
Yildiz Deniz G, Geyikoglu F, Altun S (2024) The regulatory effects of pomiferin dietary on nickel-induced hepatic injury in Sprague-Dawley rats; action mechanisms and signaling pathways. Toxicol Mech Meth 1–11. https://doi.org/10.1080/15376516.2023.2301667
Zhu Z, Pu J, Li Y, Chen J, Ding H, Zhou A, Zhang X (2023) RBM25 regulates hypoxic cardiomyocyte apoptosis through CHOP-associated endoplasmic reticulum stress. Cell Stress Chaperones 28(6):861–876. https://doi.org/10.1007/s12192-023-01380-7
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This research was funded by Natural Science Foundation of Heilongjiang Province, China (LH2023C020).
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by XL, FS, WL, YA and NS. The first draft of the manuscript was written by XL, FS, WL, YA, NS and XG and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Lv, XL., Li, WL., Sun, FJ. et al. Investigation of the mutual crosstalk between ER stress and PI3K/AKT/mTOR signaling pathway in iron overload-induced liver injury in chicks. Biometals (2024). https://doi.org/10.1007/s10534-024-00588-z
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DOI: https://doi.org/10.1007/s10534-024-00588-z