Physiological Concentration of H2O2 Supports Dopamine Neuronal Survival via Activation of Nrf2 Signaling in Glial Cells

Abstract

Traditionally, hydrogen peroxide (H2O2) was formed from cellular oxidative metabolism and often viewed as toxic waste. In fact, H2O2 was a benefit messenger for neuron-glia signaling and synaptic transmission. Thus, H2O2 was a double-edged sword and neuroprotection vs. neurotoxicity produced by H2O2 was difficult to define. Nuclear factor erythroid 2-related factor 2 (Nrf2) has been implicated as an intracellular regulator of neuronal growth. Inactivation of Nrf2 participated in the development of Parkinson's disease (PD). Thus, suitable activation of Nrf2 was essential for the prevention and treatment of PD. This study aimed to explore whether H2O2-conferred neuroprotective effects to support neuronal survival. H2O2 were added into primary neuron-glia, neuron-astroglia and neuron-microglia co-cultures in concentration- and time-dependent manners. H2O2 increased dopamine (DA) neuronal survival in concentration- and time-dependent manners. In addition, glial cells Nrf2 activation involved in H2O2-supported DA neuronal survival with the following phenomenons. First, H2O2 activated Nrf2 signaling pathway. Second, H2O2 generated beneficial neuroprotection in neuron-glia, neuron-astroglia and neuron-microglia co-cultures but not in neuron-enriched cultures. Third, silence of Nrf2 in glial cells abolished H2O2-conferred DA neuronal survival. This study demonstrated that physiological concentration of H2O2-supported DA neuronal survival via activation of Nrf2 signaling in glial cells. Our data permit to re-evaluate the role of H2O2 in the pathogenesis and therapeutic strategies for PD.

Graphic Abstract

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

ara-C:

Cytosine β-D-arabinofuranoside

CNS:

Central nervous system

DA:

Dopamine

DMF:

Dimethylfumarate

GDNF:

Glial cell-derived neurotrophic factor

HO-1:

Heme oxygenase-1

H2O2 :

Hydrogen peroxide

Keap1:

Kelch-like ECH-associated protein 1

LME:

Leu-leu methyl ester

NGF:

Nerve growth factor

Nrf2:

Nuclear factor erythroid 2-related factor 2

NQO1:

NADPH quinone oxidoreductase 1

PD:

Parkinson's disease

SNc:

Substantia nigra pars compacta

TH:

Tyrosine hydroxylase

References

  1. Ahuja M, Ammal Kaidery N, Yang L, Calingasan N, Smirnova N, Gaisin A (2016) Distinct Nrf2 signaling mechanisms of fumaric acid esters and their role in neuroprotection against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced experimental Parkinson's-like disease. J Neurosci 36:6332–6351

    CAS  Article  Google Scholar 

  2. Asanuma M, Okumura-Torigoe N, Miyazaki I, Murakami S, Kitamura Y, Sendo T (2019) Region-specific neuroprotective features of astrocytes against oxidative stress induced by 6-hydroxydopamine. Int J Mol Sci 20:pii.E598

    Article  Google Scholar 

  3. Bell KF, Al-Mubarak B, Martel MA, McKay S, Wheelan N, Hasel P (2015) Neuronal development is promoted by weakened intrinsic antioxidant defences due to epigenetic repression of Nrf2. Nat Commun 6:7066

    CAS  Article  Google Scholar 

  4. Beltrán González AN, López Pazos MI, Calvo DJ (2019) Reactive oxygen species in the regulation of the GABA mediated inhibitory neurotransmission. Neuroscience pii S0306–4522(19):30395–30401

    Google Scholar 

  5. Chen SH, Oyarzabal EA, Sung YF, Chu CH, Wang Q, Chen SL (2015) Microglial regulation of immunological and neuroprotective functions of astroglia. Glia 63:118–131

    Article  Google Scholar 

  6. Chiu SL, Cline HT (2010) Insulin receptor signaling in the development of neuronal structure and function. Neural Dev 5:7

    Article  Google Scholar 

  7. de Vries HE, Witte M, Hondius D, Rozemuller AJ, Drukarch B, Hoozemans J (2008) Nrf2-induced antioxidant protection: a promising target to counteract ROS-mediated damage in neurodegenerative disease? Free Radic Biol Med 45:1375–1383

    Article  Google Scholar 

  8. Fonseca CP, Gama S, Saavedra A, Baltazar G (2014) H2O2- or I-DOPA-injured dopaminergic neurons trigger the release of soluble mediators that up-regulate striatal GDNF through different signalling pathways. Biochim Biophys Acta 1842:927–234

    CAS  Article  Google Scholar 

  9. Gao HM, Zhou H, Zhang F, Wilson BC, Kam W, Hong JS (2011) HMGB1 acts on microglia Mac1 to mediate chronic neuroinflammation that drives progressive neurodegeneration. J Neurosci 31:1081–1092

    CAS  Article  Google Scholar 

  10. Gerich FJ, Funke F, Hildebrandt B, Fasshauer M, Müller M (2009) H2O2-mediated modulation of cytosolic signaling and organelle function in rat hippocampus. Pflugers Arch 458:937–952

    CAS  Article  Google Scholar 

  11. Groeger G, Quiney C, Cotter TG (2009) Hydrogen peroxide as a cell-survival signaling molecule. Antioxid Redox Signal 11:2655–2671

    CAS  Article  Google Scholar 

  12. Gureev AP, Popov VN (2019) Nrf2/ARE pathway as a therapeutic target for the treatment of Parkinson diseases. Neurochem Res 44:2273–2279

    CAS  Article  Google Scholar 

  13. Helou DG, Martin SF, Pallardy M, Chollet-Martin S, Kerdine-Römer S (2019) Nrf2 involvement in chemical-induced skin innate immunity. Front Immunol 10:1004

    CAS  Article  Google Scholar 

  14. Huang JY, Yuan YH, Yan JQ, Wang YN, Chu SF, Zhu CG (2016) 20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway. Acta Pharmacol Sin 37:731–740

    CAS  Article  Google Scholar 

  15. Hu XM, Zhou H, Zhang D, Yang SF, Qian L, Wu HM (2012) Clozapine protects dopaminergic neurons from inflammation-induced damage by inhibiting microglial overactivation. J Neuroimmune Pharmacol 7:187–201

    Article  Google Scholar 

  16. Jeong YH, Park JS, Kim DH, Kim HS (2016) Lonchocarpine increases Nrf2/ARE-mediated antioxidant enzyme expression by modulating AMPK and MAPK signaling in brain astrocytes. Biomol Ther 24:581–588

    CAS  Article  Google Scholar 

  17. Johnson DA, Johnson JA (2015) Nrf2-a therapeutic target for the treatment of neurodegenerative diseases. Free Radic Biol Med 88:253–267

    CAS  Article  Google Scholar 

  18. Kamsler A, Segal M (2003) Hydrogen peroxide modulation of synaptic plasticity. J Neurosci 23:269–276

    CAS  Article  Google Scholar 

  19. Lastres-Becker I, García-Yagüe AJ, Scannevin RH, Casarejos MJ, Kügler S, Rábano A (2016) Repurposing the NRF2 activator dimethyl fumarate as therapy against synucleinopathy in Parkinson's disease. Antioxid Redox Signal 25:61–77

    CAS  Article  Google Scholar 

  20. Lee CR, Patel JC, O'Neill B, Rice ME (2015) Inhibitory and excitatory neuromodulation by hydrogen peroxide: translating energetics to information. J Physiol 593:3431–3446

    CAS  Article  Google Scholar 

  21. Liddell JR, Lehtonen S, Duncan C, Keksa-Goldsteine V, Levonen AL, Goldsteins G (2016) Pyrrolidine dithiocarbamate activates the Nrf2 pathway in astrocytes. J Neuroinflammation 13:49

    Article  Google Scholar 

  22. Magalingam KB, Radhakrishnan AK, Haleagrahara N (2015) Protective mechanisms of flavonoids in Parkinson's disease. Oxid Med Cell Longev 2015:314560

    Article  Google Scholar 

  23. Machado V, Zöller T, Attaai A, Spittau B (2016) Microglia-mediated neuroinflammation and neurotrophic factor-induced protection in the MPTP mouse model of parkinson’s disease-lessons from transgenic mice. Int J Mol Sci 17:pii:E151

    Article  Google Scholar 

  24. Mimura J, Kosaka K, Maruyama A, Satoh T, Harada N, Yoshida H (2011) Nrf2 regulates NGF mRNA induction by carnosic acid in T98G glioblastoma cells and normal human astrocytes. J Biochem 150:209–217

    CAS  Article  Google Scholar 

  25. Naveilhan P, Neveu I, Jehan F, Baudet C, Wion D, Brachet P (1994) Reactive oxygen species influence nerve growth factor synthesis in primary rat astrocytes. J Neurochem 62:2178–2186

    CAS  Article  Google Scholar 

  26. Ohashi M, Hirano T, Watanabe K, Katsumi K, Ohashi N, Baba H (2016) Hydrogen peroxide modulates synaptic transmission in ventral horn neurons of the rat spinal cord. J Physiol 594:115–134

    CAS  Article  Google Scholar 

  27. Ostrowski TD, Hasser EM, Heesch CM, Kline DD (2014) H2O2 induces delayed hyperexcitability in nucleus tractus solitarii neurons. Neuroscience 262:53–69

    CAS  Article  Google Scholar 

  28. Patel JC, Rice ME (2012) Classification of H2O2 as a neuromodulator that regulates striatal dopamine release on a subsecond time scale. ACS Chem Neurosci 3:991–1001

    CAS  Article  Google Scholar 

  29. Parga JA, Rodriguez-Perez AI, Garcia-Garrote M, Rodriguez-Pallares J, Labandeira-Garcia JL (2018) Angiotensin II induces oxidative stress and upregulates neuroprotective signaling from the NRF2 and KLF9 pathway in dopaminergic cells. Free Radic Biol Med 129:394–406

    CAS  Article  Google Scholar 

  30. Persiyantseva NA, Storozhevykh TP, Senilova YE, Gorbacheva LR, Pinelis VG, Pomytkin IA (2013) Mitochondrial H2O2 as an enable signal for triggering autophosphorylation of insulin receptor in neurons. J Mol Signal 8:11

    Article  Google Scholar 

  31. Rice ME (2011) H2O2: a dynamic neuromodulator. Neuroscientist 17:389–406

    CAS  Article  Google Scholar 

  32. Shih AY, Erb H, Murphy TH (2007) Dopamine activates Nrf2-regulated neuroprotective pathways in astrocytes and meningeal cells. J Neurochem 101:109–119

    CAS  Article  Google Scholar 

  33. Schroder E, Eaton P (2008) Hydrogen peroxide as an endogenous mediator and exogenous tool in cardiovascular research: issues and considerations. Curr Opin Pharmacol 8:153–159

    Article  Google Scholar 

  34. Verkhratsky A, Nedergaard M (2018) Physiology of astroglia. Physiol Rev 98:239–389

    CAS  Article  Google Scholar 

  35. Valdovinos-Flores C, Limón-Pacheco JH, León-Rodríguez R, Petrosyan P, Garza-Lombó C (2019) Systemic L-buthionine -S-R-sulfoximine treatment increases plasma NGF and upregulates L-cys/L-cys2 transporter and γ-glutamylcysteine ligase mRNAs through the NGF/TrkA/Akt/Nrf2 pathway in the striatum. Front Cell Neurosci 13:325

    CAS  Article  Google Scholar 

  36. Wang GQ, Zhang B, He XM, Li DD, Shi JS, Zhang F (2019) Naringenin targets on astroglial Nrf2 to support dopaminergic neurons. Pharmacol Res 139:452–459

    CAS  Article  Google Scholar 

  37. Yin J, Valin KL, Dixon ML, Leavenworth JW (2017) The role of microglia and macrophages in cns homeostasis, autoimmunity, and cancer. J Immunol Res 2017:5150678

    PubMed  PubMed Central  Google Scholar 

  38. Zhang B, Wang GQ, He JY, Yang QY, Li DD, Li JJ (2019) Icariin attenuates neuroinflammation and exerts dopamine neuroprotection via an Nrf2-dependent manner. J Neuroinflammation 16:92

    CAS  Article  Google Scholar 

  39. Zhang F, Qian L, Flood PM, Shi JS, Hong JS, Gao HM (2010) Inhibition of IκB kinase-β protects dopamine neurons against lipopolysaccharide-induced neurotoxicity. J. Pharmacol. Exp. Ther. 333(3):822–833

    CAS  Article  Google Scholar 

  40. Zheng M, Liu C, Fan Y, Yan P, Shi D, Zhang Y (2017) Neuroprotection by paeoniflorin in the MPTP mouse model of Parkinson's disease. Neuropharmacology 116:412–420

    CAS  Article  Google Scholar 

Download references

Acknowledgements

All experiments were supported by National Natural Science Foundation of China (No. 81760658), High-level Innovative Talents of Guizhou Province (No. 20164027), Governor Talent Foundation of Guizhou Province (No. 201288), Innovation Research Group Project of Education Department of Guizhou Province (no. 2016038), and Excellent Young Talents of Zunyi Medical University.

Author information

Affiliations

Authors

Contributions

FZ designed all the experiments. GQW performed the experiments and wrote the manuscript. QYY, CQZ, DDL, and JJL participated in the data analysis and approved the submitted manuscript.

Corresponding author

Correspondence to Feng Zhang.

Ethics declarations

Conflicts of interest

The authors declared no conflicts of interest.

Ethics Approval

All animal experiments were performed in accordance with National Institute of Health Guideline for the Animal Care and Use of Laboratory Animal and approved protocol by the institutional Animal Care and Use Committee at Zunyi Medical University (Zunyi, China).

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wang, G., Yang, Q., Zheng, C. et al. Physiological Concentration of H2O2 Supports Dopamine Neuronal Survival via Activation of Nrf2 Signaling in Glial Cells. Cell Mol Neurobiol 41, 163–171 (2021). https://doi.org/10.1007/s10571-020-00844-z

Download citation

Keywords

  • H2O2
  • Dopamine neurons
  • Glial cells
  • Nrf2