Malaysian Healthy Ageing Society WCHA 2012 March Gamma- tocotrienol - - PowerPoint PPT Presentation

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Malaysian Healthy Ageing Society WCHA 2012 March Gamma- tocotrienol - - PowerPoint PPT Presentation

Jan 27, 2024 7 likes •209 views

Organised by: Co-Sponsored: Malaysian Healthy Ageing Society WCHA 2012 March Gamma- tocotrienol (T 3) protects human neuroblastoma SH-SY5Y cells against buthionine sulfoximine-induced cell death TAN JEN KIT Research Associate UKM

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Organised by:

Malaysian Healthy Ageing Society

Co-Sponsored:

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WCHA – 2012 March

Gamma-tocotrienol (γT3) protects human neuroblastoma SH-SY5Y cells against buthionine sulfoximine-induced cell death

TAN JEN KIT Research Associate UKM MEDICAL MOLECULAR BIOLOGY INSTITUTE

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 antioxidant

Vitamin E

Tocopherol (T)

The Encyclopedia of Vitamin E; Osakada et al. 2004

saturated 3 double bonds Tocotrienol (T3)

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 antioxidant

Vitamin E

Tocopherol (T) α- β- γ- δ-

The Encyclopedia of Vitamin E; Osakada et al. 2004

saturated 3 double bonds Methyl group (R) Tocotrienol (T3) α- β- γ- δ-

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Tocotrienols

 neuroprotection  anticancer  cardioprotective effects palm oil  gamma-tocotrienol (γT3)

(Sen et al. 2007)

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Tocotrienols

 neuroprotection  anticancer  cardioprotective effects palm oil  gamma-tocotrienol (γT3)

(Sen et al. 2007)

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Previous findings

  • γT3 protected rat astrocytes and

neuron from oxidative stress- induced apoptosis

  • Current study: to elucidate the

role of γT3-mediated apoptosis pathway in human dopaminergic neurons

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Methodology

Cell-based assays: Gene expressions Protein expressions and modification

BSO + αT BSO + γT3 αT γT3 BSO Untreated Control

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Methodology

Cell-based assays: Gene expressions Protein expressions and modifications

BSO + αT BSO + γT3 αT γT3 BSO

BSO inhibits glutathione synthesis αT as comparative isomer

Untreated Control

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Cell Viability

20 40 60 80 100 120 Cell Viability % 50 100 150 200 250 Cytotoxicity %

Cytotoxicity

50 100 150 200 250 300 Apoptosis%

Apoptosis

Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT

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Cell Viability

20 40 60 80 100 120 Cell Viability % 50 100 150 200 250 Cytotoxicity %

Cytotoxicity

50 100 150 200 250 300 Apoptosis%

Apoptosis

Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT

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Cell Viability

20 40 60 80 100 120 Cell Viability % 50 100 150 200 250 Cytotoxicity %

Cytotoxicity

50 100 150 200 250 300 Apoptosis%

Apoptosis

Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT

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Cell Viability

20 40 60 80 100 120 Cell Viability % 50 100 150 200 250 Cytotoxicity %

Cytotoxicity

50 100 150 200 250 300 Apoptosis%

Apoptosis

Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT

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Cell Viability

20 40 60 80 100 120 Cell Viability % 50 100 150 200 250 Cytotoxicity %

Cytotoxicity

50 100 150 200 250 300 Apoptosis%

Apoptosis

Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT Untreated 10uM BSO 10uM BSO 10uM BSO 100nM γT3 100nM αT

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Reactive oxygen species (ROS) detection

Untreated control 10uM BSO 10uM BSO 100nM γT3 10uM BSO 100nM αT

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0.5 1 1.5 Fold change 0.5 1 1.5 Fold change

n= 3 separate experiments * p < 0.05, significantly different from treatment with untreated control

Gene expressions

0.5 1 1.5 Fold change 0.8 0.85 0.9 0.95 1 1.05 1 2 3 4 5 6 Fold change

untreated 10uM BSO 10uM BSO 10uM BSO - - control - 100nM γT3 100nM αT 1 00nM γT3 100nM αT

Bcl2 mRNA Bax mRNA

Protein expressions

1 2 3 4 Fold change 0.5 1 1.5 Fold change

p53 Bcl2 Bax

untreated 10uM BSO 10uM BSO 10uM BSO - - control - 100nM γT3 100nM αT 100nM γT3 100nM αT

p53 mRNA

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Conclusions

not related to ROS BSO-induced cell death: p53 mRNA and Bax protein So what

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Sum-up

 γT3 and αT as potent neuroprotectants  the molecular action ? γT3 only: Bcl2 mRNA & protein Purpose?

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Acknowledgement

  • Dr. Then Sue Mian
  • Prof. Musalmah Mazlan
  • Prof. Wan Zurinah Wan Ngah

UMBI’s staffs and lab mates UKM-JJ-03-FRGS0045-2010 Thanks you

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References

Osakada, F., Hashino, A., Kume, T., Katsuki, H., Kaneko, S., & Akaike, A. 2004. α-tocotrienol provides the most potent neuroprotection among vitamin E analogs on cultured striatal neurons. Neuropharmacol. 47: 904-915. Lucarini, M. & Pedulli, G.F. 2007. Overview of antioxidant activity of viatmin e. Dlm. Preedy, V. R. & Watson, R. R. (pnyt.). The Encyclopedia of Vitamin E, hlm. 3-10. Trowbridge: CAB International. Sen, C.K., Khanna, S., and Roy, S. 2007. Tocotrinols in health and disease: the other half of the natural vitamin E family. Mol. Aspects Med. 28(5-6): 692-728. Then, SM., Mazlan, M., Top, G.M. & Wan Ngah, W.Z. 2009. Is vitamin toxic to neurons? Cell. Mol.

  • Neurobiol. 29: 485-496.