Skip to main content

Advertisement

SpringerLink
Log in

Protective effects of vitamin K2 on 6-OHDA-induced apoptosis in PC12 cells through modulation bax and caspase-3 activation

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Neuroprotection using compounds with dual functions of anti-apoptotic and antioxidant effects fight against neurodegeneration. Vitamin K2 acts as a cofactor in many biochemical pathways, including sphingolipid synthesis in the nervous system, which is involved in many cellular events, including proliferation, differentiation, cellular communication, and alteration. This study aimed to investigate the protective effects of vitamin K2 in PC12 cells as an in vitro model of Parkinson’s disease. The protective effects of vitamin K2 against 6-OHDA-induced apoptosis in PC12 cells were assessed using resazurin for viability, DCF-DA for ROS level, DTNB for glutathione level, flow cytometry for sub G1, and western blot analysis for detecting bax and pro-caspase-3 expression level. The results showed that 6-OHDA significantly decreased cell viability, glutathione and pro-caspase-3 levels, and increased ROS, the amount of bax in PC12 cells, while the pretreatment with 5 μM vitamin K2 significantly decreased the cell death induced by 6-OHDA. Generally, the results may present a new insight about the potential protective action of vitamin K2 against the progression of Parkinson’s disease. Further studies may warrant the use of vitamin K2 as an antioxidant and anti-apoptotic agent in slowing nerve injury in neurodegenerative disease, particularly in Parkinson’s disease.

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

Similar content being viewed by others

Abbreviations

Vit K2:

Vitamin K2

ROS:

Reactive oxygen species

GSH:

Glutathione

6-OHDA:

6-Hydroxydopamine

PD:

Parkinson’s disease

NGF:

Nerve growth factor

DCFH-DA:

2′, 7′-Dichlorofluorescin diacetate

DTNB:

5-5′-Dithiobis (2-nitrobenzoic acid)

PI:

Propidium iodide

NFκB:

Nuclear factor κB

ANOVA:

One-way analysis of variance

iNOS:

Inducible nitric oxide synthase

RNS:

Reactive nitrogen species

References

  1. Plaza SM, Lamson DW (2005) Vitamin K2 in bone metabolism and osteoporosis. Altern Med Rev 10:24–35

    PubMed  Google Scholar 

  2. Ferland G (2012) Vitamin K and the nervous system: an overview of its actions. Adv Nutr 3:204–212

    Article  CAS  Google Scholar 

  3. Beulens JW, Booth SL, van den Heuvel EG, Stoecklin E, Baka A, Vermeer C (2013) The role of menaquinones (vitamin K 2) in human health. Br J Nutr 110:1357–1368

    Article  CAS  Google Scholar 

  4. Ferland G (2012) Vitamin K, an emerging nutrient in brain function. BioFactors 38:151–157

    Article  CAS  Google Scholar 

  5. Iwamoto J, Sato Y, Takeda T, Matsumoto H (2012) Strategy for prevention of hip fractures in patients with Parkinson’s disease. World J Orthop 3:137–141

    Article  Google Scholar 

  6. Jankovic J (2008) Parkinson’s disease: clinical features and diagnosis. J Neurol Neurosurg Psychiatry 79:368–376

    Article  CAS  Google Scholar 

  7. Winklhofer KF, Haass C (2010) Mitochondrial dysfunction in Parkinson’s disease. Biochim Biophys Acta 1802:29–44

    Article  CAS  Google Scholar 

  8. Mousavi SH, Motaez M, Zamiri-Akhlaghi A, Emami SA, Tayarani-Najaran Z (2014) In-Vitro evaluation of cytotoxic and apoptogenic properties of Sophora Pachycarpa. Iran J Pharm Res 13:665–673

    PubMed  PubMed Central  Google Scholar 

  9. O’brien J, Wilson I, Orton T, Pognan F (2000) Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267:5421–5426

    Article  Google Scholar 

  10. Tayarani-Najaran Z, Sareban M, Gholami A, Emami SA, Mojarrab M (2013) Cytotoxic and apoptotic effects of different extracts of Artemisia turanica Krasch on K562 and HL-60 cell lines. Sci World 2013:628073–628079

    Google Scholar 

  11. Chen X, Zhong Z, Xu Z, Chen L, Wang Y (2010) 2′, 7′-Dichlorodihydrofluorescein as a fluorescent probe for reactive oxygen species measurement: forty years of application and controversy. Free Radic Res 44:587–604

    Article  CAS  Google Scholar 

  12. Lin CM, Lin RD, Chen ST, Lin YP, Chiu WT, Lin JW, Hsu FL, Lee MH (2010) Neurocytoprotective effects of the bioactive constituents of Pueraria thomsonii in 6-hydroxydopamine (6-OHDA)- treated nerve growth factor (NGF)-differentiated PC12 cells. Phytochemistry 71:2147–2156

    Article  CAS  Google Scholar 

  13. Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–169

    CAS  PubMed  Google Scholar 

  14. Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1:1458

    Article  CAS  Google Scholar 

  15. Ramazani E, Tayarani-Najaran Z, Fereidoni M (2019) Celecoxib, indomethacin and ibuprofen prevent 6-hydroxydopamine-induced PC12 cell death through the inhibition of NFκB and SAPK/JNK pathways. Iran J Basic Med Sci 22:477–484

    PubMed  PubMed Central  Google Scholar 

  16. Thijssen HH, Drittij-Reijnders MJ (1994) Vitamin K distribution in rat tissues: dietary phylloquinone is a source of tissue menaquinone-4. Br J Nutr 72:415–425

    Article  CAS  Google Scholar 

  17. Sundaram KS, Fan JH, Engelke JA, Foley AL, Suttie JW, Lev M (1996) Vitamin K status influences brain sulfatide metabolism in young mice and rats. J Nutr 126:2746–2751

    Article  CAS  Google Scholar 

  18. Crivello NA, Casseus SL, Peterson JW, Smith DE, Booth SL (2010) Age-and brain region-specific effects of dietary vitamin K on myelin sulfatides. J Nutr Biochem 21:1083–1088

    Article  CAS  Google Scholar 

  19. Blum D, Torch S, Lambeng N, Nissou MF, Benabid AL, Sadoul R, Verna JM (2001) Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson’s disease. Prog Neurobiol 65:135–172

    Article  CAS  Google Scholar 

  20. Sada E, Abe Y, Ohba R, Tachikawa Y, Nagasawa E, Shiratsuchi M, Takayanagi R (2010) Vitamin K2 modulates differentiation and apoptosis of both myeloid and erythroid lineages. Eur J Haematol 85:538–548

    Article  CAS  Google Scholar 

  21. Huang Y, Hu Z (2018) UBIAD1 protects against oxygen-glucose deprivation/reperfusion-induced multiple subcellular organelles injury through PI3K/AKT pathway in N2A cells. J Cell Physiol 233:7480–7496

    Article  CAS  Google Scholar 

  22. Furie B, Bouchard BA, Furie BC (1999) Vitamin K-dependent biosynthesis of γ-carboxyglutamic acid. Blood 93:1798–1808

    Article  CAS  Google Scholar 

  23. Hara K, Akiyama Y, Tajima T, Shiraki M (1993) Menatetrenone inhibits bone resorption partly through inhibition of PGE2 synthesis in vitro. J Bone Miner Res 8:535–542

    Article  CAS  Google Scholar 

  24. Gehrmann J, Gold R, Linington C, Lannes-Vieira J, Wekerle H, Kreutzberg GW (1993) Microglial involvement in experimental autoimmune inflammation of the central and peripheral nervous system. Glia 7:50–59

    Article  CAS  Google Scholar 

  25. Yu YX, Li YP, Gao F, Hu QS, Zhang Y, Chen D, Wang GH (2016) Vitamin K 2 suppresses rotenone-induced microglial activation in vitro. Acta Pharmacol Sin 37:1178–1189

    Article  CAS  Google Scholar 

  26. Li J, Lin JC, Wang H, Peterson JW, Furie BC, Furie B, Booth SL, Volpe JJ, Rosenberg PA (2003) Novel role of vitamin k in preventing oxidative injury to developing oligodendrocytes and neurons. J Neurosci 23:5816–5826

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Grant No. 3/42877 from the Ferdowsi university of Mashhad, Mashhad, Iran and Grant No. 951667 from the Research Affairs of Mashhad University of Medical Sciences. The authors wish to thank Mr. Malaeke for reading the flow cytometry samples.

Funding

This work has been supported by Grant No. 3/42877 from the Ferdowsi University of Mashhad, Mashhad, Iran and Grant No. 951667 from the Research Affairs of Mashhad University of Medical Sciences, Mashhad, Iran.

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

    Elham Ramazani & Masoud Fereidoni

  2. Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, P.O. Box 9188617871, Mashhad, Iran

    Zahra Tayarani-Najaran

  3. Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Zahra Tayarani-Najaran

Authors
  1. Elham Ramazani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masoud Fereidoni
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zahra Tayarani-Najaran
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ER performed the experiments and wrote the manuscript. MF and ZTN conceived, designed, and supervised the project, wrote the manuscript and provided financial support.

Corresponding author

Correspondence to Zahra Tayarani-Najaran.

Ethics declarations

Conflict of interest

There is no conflict of interest in this study.

Ethical approval

As this work is carried out in PC12 cells, there is no need for ethical clearance.

Research involving human and animal rights

Human/animals subjects were not used in the study.

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

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ramazani, E., Fereidoni, M. & Tayarani-Najaran, Z. Protective effects of vitamin K2 on 6-OHDA-induced apoptosis in PC12 cells through modulation bax and caspase-3 activation. Mol Biol Rep 46, 5777–5783 (2019). https://doi.org/10.1007/s11033-019-05011-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-019-05011-2

Keywords

Search

Navigation