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Volume 25, Issue 1
  • ISSN: 1566-5240
  • E-ISSN: 1875-5666

Abstract

Background

The pathology of Parkinson's disease (PD) indicates that iron deposition exists in dopaminergic neurons, which may be related to the death of cellular lipid iron peroxide. The extracellular autophagy adaptor SQSTM1(p62) of dopamine (DA) neurons can activate the intracellular Keap1-Nrf2-ARE signaling pathway to inhibit ferroptosis, which has a protective effect on DA neurons.

Objective

The objective of this study was to investigate the protective mechanism of the Keap1-Nrf2-ARE antioxidant pathway against iron death in dopaminergic neurons.

Methods

The experiment was divided into a control group (Control group), 1-methyl-4-phenylpyridiniumion control group (MPP+ Control group), p62 overexpression group (MPP+OV-p62), and p62 overexpression no-load group (MPP+ OV-P62-NC). The inhibitors brusatol and ZnPP inhibited the activation of NF-E2-related factor 2(Nrf2) and Heme oxygenase-1(HO-1), respectively, and were divided into brusatol group (MPP+OV-p62+brusatol) and ZnPP group (MPP+OV-p62+ZnPP). RT-qPCR was used to detect transfection efficiency, and Cell Counting Kit-8 (CCK8) was used to detect cell activity. FerroOrange, 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA), and Liperfluo probes were used to detect intracellular iron, reactive oxygen species (ROS), and lipid peroxidation (LPO) levels. Western Blotting detected the levels of Nrf2, HO-1, Kelch-like ECH-associated protein1 (Keap1), and their downstream Glutathione peroxidase 4(GPX4) and Acyl-CoA synthetase long-chain family member 4(ACSL4). The levels of L-Glutathione (GSH) and Malondialdehyde (MDA) were detected by GSH and MDA kits, and the activation of Keap1-Nrf2-ARE pathway was verified at the cellular level to have an antioxidant protective effect on iron death in dopaminergic neurons.

Results

(1) The results of RT-qPCR showed that compared with the control group, the expression of the p62 gene was significantly increased in the MPP+OV-p62 groups ( = 0.039), and the p62 gene was significantly increased in the brusatol and ZnPP groups, indicating successful transfection ( =0.002; =0.008). (2) The immunofluorescence probe flow results showed that compared to the normal control group, the contents of three kinds of probes in MPP+ model group were significantly increased ( =0.001; <0.001; <0.001), and the contents of three kinds of probes in MPP+OV-p62 group were decreased compared to the MPP+ model group ( =0.004). The results indicated that the levels of iron, ROS, and LPO were increased in the MPP+ group and decreased in the MPP+OV-p62 group. (3) Compared with the control group, the expressions of Nrf2, HO-1, and GPX4 in the MPP+OV-p62 group were increased ( =0.007; =0.004; =0.010), and the expressions of Keap1 and ACSL4 in MPP+p62 overexpression group were decreased ( =0.017; =0.005). Compared with the MPP+ control group, Nrf2 and GPX4 were increased in the MPP+OV-p62 group, and ACSL4 was decreased in the MPP+OV-p62 group ( =0.041; <0.001; <0.001). The results of the GSH and MDA kit showed that compared with the normal control group, the content of GSH in the MPP+ control group was decreased ( < 0.01), and the content of MDA was increased ( < 0.01). Compared with the MPP+ model group, GSH content was increased ( = 0.003), and MDA content was decreased in the MPP+OV-p62 group ( < 0.001). Nrf2, HO-1, and GPX4 increased in the MPP+p62 overexpression group but decreased in the brusatol group and ZnPP group ( < 0.001).

Conclusion

Based on the transfection of P62 plasmid, it was found that P62 plasmid can inhibit the lipid peroxidation of iron death in dopaminergic nerve cells by activating the Nrf2 signaling pathway, thus playing a protective role in dopaminergic nerve cells.

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2024-01-03
2025-01-19

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References

  1. Dinkova-KostovaA.T. KostovR.V. CanningP. Keap1, the cysteine-based mammalian intracellular sensor for electrophiles and oxidants.Arch. Biochem. Biophys.2017617849310.1016/j.abb.2016.08.005 27497696
     [Google Scholar]
  2. SekineH. MotohashiH. Roles of CNC transcription factors NRF1 and NRF2 in Cancer.Cancers202113354110.3390/cancers13030541 33535386
     [Google Scholar]
  3. ZhangJ. HosoyaT. MaruyamaA. Nrf2 Neh5 domain is differentially utilized in the transactivation of cytoprotective genes.Biochem. J.2007404345946610.1042/BJ20061611 17313370
     [Google Scholar]
  4. NioiP. NguyenT. SherrattP.J. PickettC.B. The carboxy-terminal Neh3 domain of Nrf2 is required for transcriptional activation.Mol. Cell. Biol.20052524108951090610.1128/MCB.25.24.10895‑10906.2005 16314513
     [Google Scholar]
  5. McMahonM. ThomasN. ItohK. YamamotoM. HayesJ.D. Redox-regulated turnover of Nrf2 is determined by at least two separate protein domains, the redox-sensitive Neh2 degron and the redox-insensitive Neh6 degron.J. Biol. Chem.200427930315563156710.1074/jbc.M403061200 15143058
     [Google Scholar]
  6. WangH. LiuK. GengM. RXRα inhibits the NRF2-ARE signaling pathway through a direct interaction with the Neh7 domain of NRF2.Cancer Res.201373103097310810.1158/0008‑5472.CAN‑12‑3386 23612120
     [Google Scholar]
  7. Lastres-BeckerI. García-YagüeA.J. ScannevinR.H. Repurposing the NRF2 activator dimethyl fumarate as therapy against synucleinopathy in Parkinson’s disease.Antioxid. Redox Signal.2016252617710.1089/ars.2015.6549 27009601
     [Google Scholar]
  8. PrasadS. SajjaR.K. KaisarM.A. Role of Nrf2 and protective effects of Metformin against tobacco smoke-induced cerebrovascular toxicity.Redox Biol.201712586910.1016/j.redox.2017.02.007 28212524
     [Google Scholar]
  9. JiY.J. WangH.L. YinB.L. RenX.Y. Down-regulation of DJ-1 augments neuroinflammation via Nrf2/Trx1/NLRP3 axis in MPTP-induced Parkinson’s disease mouse model.Neuroscience202044225326310.1016/j.neuroscience.2020.06.001 32526245
     [Google Scholar]
  10. WangQ. BotchwayB.O.A. ZhangY. LiuX. Ellagic acid activates the Keap1-Nrf2-ARE signaling pathway in improving Parkinson’s disease: A review.Biomed. Pharmacother.202215611384810.1016/j.biopha.2022.113848 36242848
     [Google Scholar]
  11. LuX.Y. WangH.D. XuJ.G. DingK. LiT. Deletion of Nrf2 exacerbates oxidative stress after traumatic brain injury in mice.Cell. Mol. Neurobiol.201535571372110.1007/s10571‑015‑0167‑9 25732597
     [Google Scholar]
  12. LiX. YingH. ZhangZ. Sulforaphane attenuates chronic intermittent hypoxia-induced brain damage in mice via augmenting Nrf2 nuclear translocation and autophagy.Front. Cell. Neurosci.20221682752710.3389/fncel.2022.827527 35401114
     [Google Scholar]
  13. SharmaV. KaurA. SinghT.G. Counteracting role of nuclear factor erythroid 2-related factor 2 pathway in Alzheimer’s disease.Biomed. Pharmacother.202012911037310.1016/j.biopha.2020.110373 32603894
     [Google Scholar]
  14. HirotsuY. KatsuokaF. FunayamaR. Nrf2–MafG heterodimers contribute globally to antioxidant and metabolic networks.Nucleic Acids Res.20124020102281023910.1093/nar/gks827 22965115
     [Google Scholar]
  15. ValionyteE. BarrowE.R. BaxterC.R. LuoS. A dominant-negative regulatory mechanism of SQSTM1 droplets-based autophagy.Autophagy202218493593610.1080/15548627.2022.2029672 35188067
     [Google Scholar]
  16. ChenJ. ZhangJ. ChenT. Xiaojianzhong decoction attenuates gastric mucosal injury by activating the p62/Keap1/Nrf2 signaling pathway to inhibit ferroptosis.Biomed. Pharmacother.202215511363110.1016/j.biopha.2022.113631 36122518
     [Google Scholar]
  17. XuY. LiY. LiJ. ChenW. Ethyl carbamate triggers ferroptosis in liver through inhibiting GSH synthesis and suppressing Nrf2 activation.Redox Biol.20225310234910.1016/j.redox.2022.102349 35623314
     [Google Scholar]
  18. YuG. DengA. TangW. MaJ. YuanC. MaJ. Hydroxytyrosol induces phase II detoxifying enzyme expression and effectively protects dopaminergic cells against dopamine- and 6-hydroxydopamine induced cytotoxicity.Neurochem. Int.20169611312010.1016/j.neuint.2016.03.005 26970393
     [Google Scholar]
  19. NeisV.B. RosaP.B. MorettiM. RodriguesA.L.S. Involvement of heme oxygenase-1 in neuropsychiatric and neurodegenerative diseases.Curr. Pharm. Des.201824202283230210.2174/1381612824666180717160623 30019638
     [Google Scholar]
  20. BianY. ChenY. WangX. Oxyphylla A ameliorates cognitive deficits and alleviates neuropathology via the Akt-GSK3β and Nrf2-Keap1-HO-1 pathways in vitro and in vivo murine models of Alzheimer’s disease.J. Adv. Res.20213411210.1016/j.jare.2021.09.002 35024177
     [Google Scholar]
  21. Abu-ElfotuhK. HamdanA.M.E. MohammedA.A. Neuroprotective effects of some nutraceuticals against manganese-induced Parkinson’s disease in rats: possible modulatory effects on TLR4/NLRP3/NF-κB, GSK-3β, Nrf2/HO-1, and apoptotic pathways.Pharmaceuticals20221512155410.3390/ph15121554 36559006
     [Google Scholar]
  22. XuJ. XiaoC. SongW. Elevated heme oxygenase-1 correlates with increased brain iron deposition measured by quantitative susceptibility mapping and decreased hemoglobin in patients with Parkinson’s disease.Front. Aging Neurosci.20211365662610.3389/fnagi.2021.656626 33815094
     [Google Scholar]
  23. ZhangF. YanY. PengW. PARK7 promotes repair in early steroid-induced osteonecrosis of the femoral head by enhancing resistance to stress-induced apoptosis in bone marrow mesenchymal stem cells via regulation of the Nrf2 signaling pathway.Cell Death Dis.2021121094010.1038/s41419‑021‑04226‑1 34645791
     [Google Scholar]
/content/journals/cmm/10.2174/0115665240266555231120044938
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A Mechanism Study on the Antioxidant Pathway of Keap1-Nrf2-ARE Inhibiting Ferroptosis in Dopaminergic Neurons
Current Molecular Medicine 25, 37 (2025); https://doi.org/10.2174/0115665240266555231120044938
/content/journals/cmm/10.2174/0115665240266555231120044938
/content/journals/cmm/10.2174/0115665240266555231120044938
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  • Article Type:     Research Article
Keyword(s): antioxidant damage; iron death; nuclear transcription factor 2; Parkinson's disease; pathogenesis; regulatory pathway

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