Skip to main content
SpringerLink
Log in

NLRP3 Inflammasome Inhibition Prevents α-Synuclein Pathology by Relieving Autophagy Dysfunction in Chronic MPTP–Treated NLRP3 Knockout Mice

  • Published:
Molecular Neurobiology Aims and scope Submit manuscript

Abstract

Recent researches showed that nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) inflammasome inhibition exerted dopaminergic neuroprotection in cellular or animal models of Parkinson’s disease (PD). NLRP3 inflammasome has been proposed as a drug target for treatment of PD. However, the interplay between chronic NLRP3 inflammasome and progressive α-synuclein pathology keeps poorly understood. Moreover, the potential mechanism keeps unknown. In the present study, we investigate whether NLRP3 inflammasome inhibition prevents α-synuclein pathology by relieving autophagy dysfunction in the chronic 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) mouse model of PD. NLRP3 knockout mice and their wild-type counterparts were treated with continuous MPTP administration via osmotic mini-pumps. Dopaminergic neuronal degeneration was assessed by western blotting and immunohistochemistry (IHC). The levels of dopamine and its metabolites were determined using high-performance liquid chromatography. NLRP3 inflammasome activation and autophagy biomarkers were assessed by western blot. The expressions of pro-inflammatory cytokines were measured by ELISA. The glial reaction and α-synuclein pathology were assessed by IHC and immunofluorescence. Our results show that NLRP3 inflammasome inhibition via NLRP3 knockout not only protects against nigral dopaminergic degeneration and striatal dopamine deletion but also prevents nigral pathological α-synuclein formation in PD mice. Furthermore, it significantly suppresses MPTP-induced glial reaction accompanied by the secretion of pro-inflammatory cytokines in the midbrain of mice. Most importantly, it relieves autophagy dysfunction in the midbrain of PD mice. Collectively, we demonstrate for the first time that improving autophagy function is involved in the preventive effect of NLRP3 inflammasome inhibition on α-synuclein pathology in PD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Hirsch EC, Vyas S, Hunot S (2012) Neuroinflammation in Parkinson's disease. Parkinsonism Relat Disord 18(Suppl 1):S210–S212

    Article  Google Scholar 

  2. Heneka MT, McManus RM, Latz E (2018) Inflammasome signalling in brain function and neurodegenerative disease. Nat Rev Neurosci 19(10):610–621

    Article  CAS  Google Scholar 

  3. Gordon R, Albornoz EA, Christie DC et al (2018) Inflammasome inhibition prevents α-synuclein pathology and dopaminergic neurodegeneration in mice. Sci Transl Med 10(465):eaah4066

  4. Mao Z, Liu C, Ji S, Yang Q, Ye H, Han H, Xue Z (2017) The NLRP3 Inflammasome is involved in the pathogenesis of Parkinson's disease in rats. Neurochem Res 42(4):1104–1115

    Article  CAS  Google Scholar 

  5. Sarkar S, Malovic E, Harishchandra DS, Ghaisas S, Panicker N, Charli A, Palanisamy BN, Rokad D et al (2017) Mitochondrial impairment in microglia amplifies NLRP3 inflammasome proinflammatory signaling in cell culture and animal models of Parkinson's disease. NPJ Parkinsons Dis 3:30

  6. Lee E, Hwang I, Park S, Hong S, Hwang B, Cho Y, Son J, Yu JW (2019) MPTP-driven NLRP3 inflammasome activation in microglia plays a central role in dopaminergic neurodegeneration. Cell Death Differ 26(2):213–228

  7. Yan Y, Jiang W, Liu L, Wang X, Ding C, Tian Z, Zhou R (2015) Dopamine controls systemic inflammation through inhibition of NLRP3 inflammasome. Cell. 160(1–2):62–73

    Article  CAS  Google Scholar 

  8. Han X, Sun S, Sun Y, Song Q, Zhu J, Song N, Chen M, Sun T et al (2019) Small molecule-driven NLRP3 inflammation inhibition via interplay between ubiquitination and autophagy: Implications for Parkinson disease. Autophagy. 15(11):1860–1881

    Article  CAS  Google Scholar 

  9. Hou X, Watzlawik JO, Fiesel FC, Springer W (2020) Autophagy in Parkinson’s disease. J Mol Biol 432(8):2651–2672

  10. Martins JD, Liberal J, Silva A, Ferreira I, Neves BM, Cruz MT (2015) Autophagy and inflammasome interplay. DNA Cell Biol 34(4):274–281

    Article  CAS  Google Scholar 

  11. Fornai F, Schlüter OM, Lenzi P et al (2005) Parkinson-like syndrome induced by continuous MPTP infusion: Convergent roles of the ubiquitin-proteasome system and alpha-synuclein. Proc Natl Acad Sci U S A 102(9):3413–3418

    Article  CAS  Google Scholar 

  12. Gibrat C, Saint-Pierre M, Bousquet M, Lévesque D, Rouillard C, Cicchetti F (2009) Differences between subacute and chronic MPTP mice models: Investigation of dopaminergic neuronal degeneration and alpha-synuclein inclusions. J Neurochem 109(5):1469–1482

    Article  CAS  Google Scholar 

  13. Wu KC, Liou HH, Lee CY, Lin CJ (2019) Down-regulation of natural resistance-associated macrophage protein-1 (Nramp1) is associated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)/1-methyl-4-phenylpyridinium (MPP(+) )-induced alpha-synuclein accumulation and neurotoxicity. Neuropathol Appl Neurobiol 45(2):157–173

    Article  CAS  Google Scholar 

  14. Pitcairn C, Wani WY, Mazzulli JR (2019) Dysregulation of the autophagic-lysosomal pathway in Gaucher and Parkinson's disease. Neurobiol Dis 122:72–82

    Article  CAS  Google Scholar 

  15. Alvarez-Fischer D, Guerreiro S, Hunot S, Saurini F, Marien M, Sokoloff P, Hirsch EC, Hartmann A et al (2008) Modelling Parkinson-like neurodegeneration via osmotic minipump delivery of MPTP and probenecid. J Neurochem 107(3):701–711

  16. Sekar S, Mani S, Rajamani B, Manivasagam T, Thenmozhi AJ, Bhat A, Ray B, Essa MM et al (2018) Telmisartan ameliorates Astroglial and dopaminergic functions in a mouse model of chronic parkinsonism. Neurotox Res 34(3):597–612

  17. Chen L, Xue L, Zheng J, Tian X, Zhang Y, Tong Q (2019) PPARss/delta agonist alleviates NLRP3 inflammasome-mediated neuroinflammation in the MPTP mouse model of Parkinson's disease. Behav Brain Res 356:483–489

    Article  CAS  Google Scholar 

  18. Muñoz-Manchado AB, Villadiego J, Romo-Madero S, Suárez-Luna N, Bermejo-Navas A, Rodríguez-Gómez JA, Garrido-Gil P, Labandeira-García JL et al (2016) Chronic and progressive Parkinson's disease MPTP model in adult and aged mice. J Neurochem 136(2):373–387

  19. Burks S, Raymick J, Robinson B, Hanig J, Sarkar S (2019) Neuroprotective effects of acetyl-l-carnitine (ALC) in a chronic MPTP-induced Parkinson's disease mouse model: Endothelial and microglial effects. Neurosci Lett 703:86–95

    Article  CAS  Google Scholar 

  20. Gustin A, Kirchmeyer M, Koncina E, Felten P, Losciuto S, Heurtaux T, Tardivel A, Heuschling P et al (2015) NLRP3 Inflammasome is expressed and functional in mouse brain microglia but not in astrocytes. PLoS One 10(6):e0130624

  21. Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell. 140(3):313–326

    Article  CAS  Google Scholar 

  22. Bjørkøy G, Lamark T, Pankiv S, Øvervatn A, Brech A, Johansen T (2009) Monitoring autophagic degradation of p62/SQSTM1. Methods Enzymol 452:181–197

    Article  Google Scholar 

  23. Karabiyik C, Lee MJ, Rubinsztein DC (2017) Autophagy impairment in Parkinson's disease. Essays Biochem 61(6):711–720

    Article  Google Scholar 

  24. Niranjan R, Mishra KP, Thakur AK (2018) Inhibition of Cyclooxygenase-2 (COX-2) initiates autophagy and potentiates MPTP-induced Autophagic cell death of human Neuroblastoma cells, SH-SY5Y: An inside in the pathology of Parkinson's disease. Mol Neurobiol 55(10):8038–8050

    Article  CAS  Google Scholar 

  25. Yuk JM, Jo EK (2013) Crosstalk between autophagy and inflammasomes. Mol Cell 36(5):393–399

    Article  CAS  Google Scholar 

  26. François A, Terro F, Janet T, Rioux Bilan A, Paccalin M, Page G (2013) Involvement of interleukin-1β in the autophagic process of microglia: Relevance to Alzheimer's disease. J Neuroinflammation 10:151

    Article  Google Scholar 

Download references

Funding

This study was supported by grants from the National Natural Science Foundation of China (No. 81600981), the Natural Science Foundation of Jiangsu Province (BK20191212), the Six Talent Peaks Project in Jiangsu Province (WSN-282), and the 533 Talent Project of Huai’an City (HAA201749).

Author information

Author notes

  1. Zhou Ou, Yuanzhang Zhou and Lijun Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, The Affiliated Huaian No. 1 People’s Hospital of Nanjing Medical University, 1 Huanghe Road West, Huaian, 223300, Jiangsu, China

    Zhou Ou, Lijun Wang, Liujun Xue, Jinlong Zheng & Qiang Tong

  2. Jiangsu Key Laboratory of Neurodegeneration, Department of Pharmacology, Nanjing Medical University, 101 Longmian Avenue, Nanjing, 211166, Jiangsu, China

    Yuanzhang Zhou

  3. Division of Neuropathology, Department of Pathology, Johns Hopkins University School of Medicine, 720 Rutland Ave, Baltimore, MD, 21205, USA

    Liam Chen & Qiang Tong

Authors
  1. Zhou Ou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuanzhang Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lijun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liujun Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jinlong Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Liam Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qiang Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Q. Tong conceived and designed the experiments. Z. Ou and Y.Z. Zhou performed primary experiments. L.J. Wang performed the data analyses and supplementary experiments during the process of revising the manuscript. L.J. Xue performed the data analyses. J.L. Zheng helped to perform the analysis with constructive discussions. Q. Tong wrote the manuscript. L. Chen helped to perform the analysis with constructive discussions and revised the manuscript.

Corresponding author

Correspondence to Qiang Tong.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s Note

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

Supplementary Information

ESM 1

(DOCX 194 kb).

ESM 2

(DOCX 304 kb).

ESM 3

(XLSX 17 kb).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ou, Z., Zhou, Y., Wang, L. et al. NLRP3 Inflammasome Inhibition Prevents α-Synuclein Pathology by Relieving Autophagy Dysfunction in Chronic MPTP–Treated NLRP3 Knockout Mice. Mol Neurobiol 58, 1303–1311 (2021). https://doi.org/10.1007/s12035-020-02198-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12035-020-02198-5

Keywords

Search

Navigation