Skip to main content
SpringerLink
Log in

Tempol and silymarin rescue from zinc-induced degeneration of dopaminergic neurons through modulation of oxidative stress and inflammation

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Oxidative stress and inflammation are the key players in the toxic manifestation of sporadic Parkinson’s disease and zinc (Zn)-induced dopaminergic neurodegeneration. A synthetic superoxide dismutase (SOD) mimetic, tempol, and a naturally occurring antioxidant, silymarin protect against oxidative stress-mediated damage. The study intended to explore the effects of tempol and silymarin against Zn-induced dopaminergic neurodegeneration. Exposure to Zn produced neurobehavioral deficits and striatal dopamine depletion. Zn reduced glutathione content and glutathione-S-transferase activity and increased lipid peroxidation, superoxide dismutase activity, and level of pro-inflammatory mediators [nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6)]. Zn also attenuated the expression of tyrosine hydoxylase (TH), vesicular monoamine transporter 2 (VMAT-2), mitochondrial B-cell lymphoma-2 (Bcl-2), and procaspase-3 and 9 proteins and number of TH-positive neurons. Conversely, Zn elevated the expression of dopamine transporter (DAT) and mitochondrial Bcl-2-associated X (Bax) protein along with mitochondrial cytochrome c release. Administration of tempol significantly alleviated Zn-induced motor impairments, dopamine depletion, reduction in TH expression, and loss of TH-positive neurons similar to silymarin. Silymarin mitigated Zn-induced oxidative stress and inflammation and restored the expression of dopamine transporters and levels of pro-apoptotic proteins akin to tempol. The results demonstrate that both tempol and silymarin protect against Zn-induced dopaminergic neuronal loss through the suppression of oxidative stress and inflammation.

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
Fig. 7

Similar content being viewed by others

Data availability

All the experimental data generated or analyzed during the present study are available from the corresponding author upon genuine request.

References

  1. Simon DK, Tanner CM, Brundin P (2020) Parkinson disease epidemiology, pathology, genetics, and pathophysiology. Clin Geriatr Med 36(1):1–12

    Article  PubMed  Google Scholar 

  2. Goldman SM (2014) Environmental toxins and Parkinson’s disease. Annu Rev Pharmacol Toxicol 54:141–164

    Article  CAS  PubMed  Google Scholar 

  3. Bjorklund G, Stejskal V, Urbina MA, Dadar M, Chirumbolo S, Mutter J (2018) Metals and parkinson’s disease: mechanisms and biochemical processes. Curr Med Chem 25(19):2198–2214

    Article  CAS  PubMed  Google Scholar 

  4. Dexter DT, Carayon A, Javoy-Agid F, Agid Y, Wells FR, Daniel SE, Lees AJ, Jenner P, Marsden CD (1991) Alterations in the levels of iron, ferritin and other trace metals in Parkinson’s disease and other neurodegenerative diseases affecting the basal ganglia. Brain A J Neurol 114:1953–1975

    Article  Google Scholar 

  5. Kumar A, Singh BK, Ahmad I, Shukla S, Patel DK, Srivastava G, Kumar V, Pandey HP, Singh C (2012) Involvement of NADPH oxidase and glutathione in zinc-induced dopaminergic neurodegeneration in rats: similarity with paraquat neurotoxicity. Brain Res 1438:48–64

    Article  CAS  PubMed  Google Scholar 

  6. Kumar V, Singh BK, Chauhan AK, Singh D, Patel DK, Singh C (2016) Minocycline rescues from zinc-induced nigrostriatal dopaminergic neurodegeneration: biochemical and molecular interventions. Mol Neurobiol 53(5):2761–2777

    Article  CAS  PubMed  Google Scholar 

  7. Chauhan AK, Mittra N, Kumar V, Patel DK, Singh C (2016) Inflammation and B-cell lymphoma-2 associated X protein regulate zinc-induced apoptotic degeneration of rat nigrostriatal dopaminergic neurons. Mol Neurobiol 53:5782–5795

    Article  CAS  PubMed  Google Scholar 

  8. Park JS, Blair NF, Sue CM (2015) The role of ATP13A2 in Parkinson’s disease: clinical phenotypes and molecular mechanisms. Mov Disord 30(6):770–779

    Article  CAS  PubMed  Google Scholar 

  9. Takeda A, Tamano H (2020) New insight into Parkinson’s disease pathogenesis from reactive oxygen species-mediated extracellular Zn2+ influx. J Trace Elem Med Biol 61:126545

    Article  Google Scholar 

  10. Sikora J, Kieffer BL, Paoletti P, Ouagazzal AM (2020) Synaptic zinc contributes to motor and cognitive deficits in 6-hydroxydopamine mouse models of Parkinson’s disease. Neurobiol Dis 134:104681

    Article  CAS  PubMed  Google Scholar 

  11. Taylor JM, Main BS, Crack PJ (2013) Neuroinflammation and oxidative stress: co-conspirators in the pathology of Parkinson’s disease. Neurochem Int 62(5):803–819

    Article  CAS  PubMed  Google Scholar 

  12. Trist BG, Hare DJ, Double KL (2019) Oxidative stress in the aging substantia nigra and the etiology of Parkinson’s disease. Aging Cell 18(6):e13031

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Tansey MG, Goldberg MS (2010) Neuroinflammation in Parkinson’s disease: its role in neuronal death and implications for therapeutic intervention. Neurobiol Dis 37(3):510–518

    Article  CAS  PubMed  Google Scholar 

  14. Badanjak K, Fixemer S, Smajic S, Skupin A, Grunewald A (2021) The contribution of microglia to neuroinflammation in Parkinson’s disease. Int J Mol Sci 22:4676

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Dixit A, Srivastava G, Verma D, Mishra M, Singh PK, Prakash O (1832) Singh MP (2013) Minocycline, levodopa and MnTMPyP induced changes in the mitochondrial proteome profile of MPTP and maneb and paraquat mice models of Parkinson’s disease. Biochim Biophys Acta 8:1227–1240

    Google Scholar 

  16. Agrawal S, Singh A, Tripathi P, Mishra M, Singh PK, Singh MP (2015) Cypermethrin-induced nigrostriatal dopaminergic neurodegeneration alters the mitochondrial function: a proteomics study. Mol Neurobiol 51(2):448–465

    Article  CAS  PubMed  Google Scholar 

  17. Shibata H, Katsuki H, Okawara M, Kume T, Akaike A (2006) c-Jun N-terminal kinase inhibition and alpha-tocopherol protect dopaminergic neurons from interferon-gamma/lipopolysaccharide-induced injury without affecting nitric oxide production. J Neurosci Res 83(1):102–109

    Article  CAS  PubMed  Google Scholar 

  18. Singhal NK, Srivastava G, Patel DK, Jain SK, Singh MP (2011) Melatonin or silymarin reduces maneb and paraquat-induced Parkinson’s disease phenotype in the mouse. J Pineal Res 50(2):97–109

    CAS  PubMed  Google Scholar 

  19. Wang MJ, Lin WW, Chen HL, Chang YH, Ou HC, Kuo JS, Hong JS, Jeng KCG (2002) Silymarin protects dopaminergic neurons against lipopoloysaccharide-induced neurotoxicity by inhibiting microglia activation. Eur J Neurosci 16:2103–2112

    Article  PubMed  Google Scholar 

  20. Haddadi R, Nayebi AM, Farajniya S, Brooshghalan SE, Sharifi H (2014) Silymarin improved 6-OHDA-induced motor impairment in hemi-Parkinsonian rats: behavioural and molecular study. Daru J Pharm Sci 22:38

    Article  Google Scholar 

  21. Tripathi MK, Rasheed MSU, Mishra AK, Patel DK, Singh MP (2020) Silymarin protects against impaired autophagy associated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinsonism. J Mol Neurosci 70(2):276–283

    Article  CAS  PubMed  Google Scholar 

  22. Wilcox CS (2010) Effects of tempol and redox-cycling nitroxides in models of oxidative stress. Pharmacol Ther 126(2):119–145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Solesio ME, Saez-Atienzar S, Jordan J, Galindo MF (2012) Characterization of mitophagy in the 6-hydroxydopamine Parkinson’s disease model. Toxicol Sci 129(2):411–420

    Article  CAS  PubMed  Google Scholar 

  24. Chauhan AK, Mittra N, Patel DK, Singh C (2018) Cyclooxygenase-2 directs microglial activation-mediated inflammation and oxidative stress leading to intrinsic apoptosis in Zn-induced Parkinsonism. Mol Neurobiol 55(3):2162–2173

    Article  CAS  PubMed  Google Scholar 

  25. Ahmad I, Shukla S, Kumar A, Singh BK, Kumar V, Chauhan AK, Singh D, Pandey HP, Singh C (2013) Biochemical and molecular mechanisms of N-acetyl cysteine and silymarin-mediated protection against maneb- and paraquat-induced hepatotoxicity in rats. Chem Biol Interact 201(1–3):9–18

    Article  CAS  PubMed  Google Scholar 

  26. Singh BK, Kumar A, Ahmad I, Kumar V, Patel DK, Jain SK, Singh C (2011) Oxidative stress in zinc-induced dopaminergic neurodegeneration: implications of superoxide dismutase and heme oxygenase-1. Free Radic Res 45(10):1207–1222

    Article  CAS  PubMed  Google Scholar 

  27. Kumar A, Ahmad I, Shukla S, Singh BK, Patel DK, Pandey HP, Singh C (2010) Effect of zinc and paraquat co-exposure on neurodegeneration: modulation of oxidative stress and expression of metallothionein’s, toxicant responsive and transporter genes in rats. Free Radical Res 44:950–965

    Article  CAS  Google Scholar 

  28. Mittra N, Chauhan AK, Singh G, Patel DK, Singh C (2020) Postnatal zinc or paraquat administration increases paraquat or zinc-induced loss of dopaminergic neurons: insight into augmented neurodegeneration. Mol Cell Biochem 467(1–2):27–43

    Article  CAS  PubMed  Google Scholar 

  29. Portbury SD, Adlard PA (2017) Zinc signal in brain diseases. Int J Mol Sci 18:2506

    Article  PubMed  PubMed Central  Google Scholar 

  30. Liu X, Liu W, Wang C, Chen Y, Liu P, Hayashi T, Mizuno K, Hattori S, Fujisaki H, Ikejima T (2021) Silibinin attenuates motor dysfunction in a mouse model of Parkinson’s disease by suppression of oxidative stress and neuroinflammation along with promotion of mitophagy. Physiol Behav 239:113510

    Article  CAS  PubMed  Google Scholar 

  31. Haddadi R, Nayebi AM, Brooshghalan, (2018) Silymarin prevents apoptosis through inhibiting the Bac/caspase-3 expression and suppresses toll-like receptor-4 pathway in the SNc of 6-OHDA intoxicated rats. Biomed Pharmacother 104:127–136

    Article  CAS  PubMed  Google Scholar 

  32. Liang Q, Smith AD, Pan S, Tyurin VA, Kagan VE, Hastings TG, Schor NF (2005) Neuroprotective effects of TEMPOL in central and peripheral nervous system models of Parkinson’s disease. Biochem Pharmacol 70(9):1371–1381

    Article  CAS  PubMed  Google Scholar 

  33. Pérez-H J, Carrillo-S C, García E, Ruiz-Mar G, Pérez-Tamayo R, Chavarria A (2014) Neuroprotective effect of silymarin in a MPTP mouse model of Parkinson’s disease. Toxicology 319:38–43

    Article  PubMed  Google Scholar 

  34. Sunkaria A, Sharma DR, Wani WY, Gill KD (2014) Attenuation of dichlorvos-induced microglial activation and neuronal apoptosis by 4-hydroxy TEMPO. Mol Neurobiol 49(1):163–175

    Article  CAS  PubMed  Google Scholar 

  35. Dohare P, Hyzinski-Garcia MC, Vipani A, Bowens NH, Nalwalk JW, Feustel PJ, Keller RW Jr, Jourd’heuil D, Mongin AA, (2014) The neuroprotective properties of the superoxide dismutase mimetic tempol correlate with its ability to reduce pathological glutamate release in a rodent model of stroke. Free Radic Biol Med 77:168–182

    Article  CAS  PubMed  Google Scholar 

  36. Morgan MJ, Liu ZG (2011) Crosstalk of reactive oxygen species and NF-κB signalling. Cell Res 21(1):103–115

    Article  CAS  PubMed  Google Scholar 

  37. Glass CK, Saijo K, Winner B, Marchetto MC, Gage FH (2010) Mechanisms underlying inflammation in neurodegeneration. Cell 140(6):918–934

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. He J, Zhu G, Wang G, Zhang F (2020) Oxidative stress and neuroinflammation potentiate each other to promote progression of dopamine neurodegeneration. Oxid Med Cell Long 6137521:12

    Google Scholar 

  39. Haddadi R, Nayebi AM, Brooshghalan SE (2013) Pre-treatment with silymarin reduces brain myeloperoxidase activity and inflammatory cytokines in 6-OHDA hemi-parkinsonian rats. Neurosci Lett 555:106–111

    Article  CAS  PubMed  Google Scholar 

  40. Singhal NK, Chauhan AK, Jain SK, Shanker R, Singh C, Singh MP (2013) Silymarin- and melatonin-mediated changes in the expression of selected genes in pesticides-induced Parkinsonism. Mol Cell Biochem 384(1–2):47–58

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are genuinely thankful to Department of Science & Technology (DST), New Delhi, India for granting the financial aid as research fellowships to Garima Singh and Namrata Mittra. The CSIR-IITR communication number allotted to this article is IITR/SEC/2022-2023/15.

Funding

The authors have not disclosed any funding.

Author information

Authors and Affiliations

  1. Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Vishvigyan Bhawan, 31, Mahatma Gandhi Marg, Post Box No. 80, Lucknow, 226 001, UP, India

    Garima Singh, Namrata Mittra & Chetna Singh

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201 002, UP, India

    Garima Singh, Namrata Mittra & Chetna Singh

Authors
  1. Garima Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Namrata Mittra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chetna Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

GS mainly carried out the experimental work, data compilation, and analysis. She also wrote the initial draft of the manuscript. NM was involved in the estimation of neurotransmitters and also assisted in Western blotting of few proteins, data compilation, and analysis. CS contributed to study conceptualization, experimental design, and reviewing/editing of the text of the first draft provided by GS. The final version of the manuscript has been read and approved by all the authors.

Corresponding author

Correspondence to Chetna Singh.

Ethics declarations

Conflict of interest

No conflict of interest is affirmed by the authors for this study.

Ethical approval

The animal experimentation was carried out following the approval by Institutional Animal Ethics Committee of CSIR-IITR (Reference number: IITR/IAEC/89/17–23/19–33/20) in accord with the standard ethical guidelines for animal use.

Additional information

Publisher's Note

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

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Singh, G., Mittra, N. & Singh, C. Tempol and silymarin rescue from zinc-induced degeneration of dopaminergic neurons through modulation of oxidative stress and inflammation. Mol Cell Biochem 478, 1705–1718 (2023). https://doi.org/10.1007/s11010-022-04620-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-022-04620-z

Keywords

Search

Navigation