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Biological Activities of Resveratrol
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Biological activities of resveratrol
1 The research program: cell and molecular biology of longevity1. The research program: cell and molecular biology of longevity
2. Biological activities of resveratrol in mammals (including humans)
Animal lifespans
S l d s CenturiesSeveral days Centuries
lifespan Harriet, who died on Friday, June 23, 2006 at the age of 176 years is said to be the
The mayflyLifespan: ~ 1day!
oldest animal in captivity, according to the Guinness Book of Records.(AP/Australia Zoo)
y = 6.4471x0.138
R2 0 3395
1000
R2 = 0.3395
100(y
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11 100 10000 1000000 100000000
Log Adult Mass (g)Log Adult Mass (g)
Why?
How?
Cells of longer-lived animals are
hard to kill
- increased cellular t i t i stress resistance in
longer-lived animals
Stress resistance of skin fibroblasts correlates with species MLSP
Kapahi et al., 1999, Free Radic. Biol. Med.
Long-lived Snell dwarf mice have t i t t fib bl tstress resistant fibroblasts
Snell & normal mouse lifespan plot Snell cells resistant to stressors
Murakami et al., Am. J. Physiol. 2003
Species-omics
Identifying cellular characteristics that co-evolve with longevity and might explain stress resistanceg p
Molecular mechanisms underlying longevity in animals
Two experimental models:Two experimental models:
(1) The long-lived Snell dwarf mouse
(2) A comparative model using mammalian and avian species(2) A comparative model using mammalian and avian species
What have we learned?
C ll l h t isti s C ll l h t isti s Cellular characteristics that do not correlate,
or correlate negatively, with longevity
Cellular characteristics that correlate
positively with longevitywith longevity
12 112 1
MnSOD correlates with longevity
Dermal fibroblasts
MnSOD correlates with longevity
mammalian brain tissue
MnSOD prevents cytochrome c release during cell stress
Cytochrome c is a key cell death signal
Ellen Robb, PhD candidate
Resveratrol does everything!
Anti-cancer activity
Anti-oxidant
Anti aging Anti-aging
Neuroprotective
What is the effect on human cells of long-term exposure to resveratrol?
No effect of resveratrol on most antioxidant enzymes
t li id di te.g. cytosolic superoxide dismutase
0, 50 or 100 M trans-resveratrol
But, a huge effect on MnSOD!
0, 50 or 100 M trans-resveratrol
MnSOD effect observed in a wide variety of cell typese
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myoblasts Neuroblastoma cells
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24h 72h24h 72h 1wk
0, 50 or 100 M trans-resveratrol
Proof that this matters?
Greater MnSOD levels paralleled by greater cellular stress resistance
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ResveratrolControl
Figure 1: Chronicexposureofmusclecells,lungcellsandneuralcellsto50Mresveratrolaffordsprotectionagainstcelldeathinducedbytheoxidativestressorsp g yhydrogenperoxideandparaquat.DatapointrepresentsLD50foragiventoxin,Cellsurvivalwasevaluatedbytrypanblueexclusion.Datarepresentsthemeanof5independentmeasurements SEM.*=P
Greater MnSOD levels paralleled by greater cellular stress resistance
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Figure 1: Chronicexposureofmusclecells,lungcellsandneuralcellsto50Mresveratrolaffordsprotectionagainstcelldeathinducedbytheoxidativestressorshydrogenperoxideandparaquat.DatapointrepresentsLD50foragiventoxin,Cellsurvivalwasevaluatedbytrypanblueexclusion.Datarepresentsthemeanof5independentmeasurements SEM.*=P
Greater MnSOD levels paralleled by reduced replicative growth rate
50
Human lung fibroblast cells
35404550
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ControlRES
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Time (hours)
How is resveratrol regulating MnSOD levels?
Resveratrol is a phytoestrogen
R t lResveratrol
Estrogen (estradiol) elicits a similar MnSOD effect
4.5
3.5
4
4.5
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3 DMSO
50uM RES
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2 100nM estradiol
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Lung fibroblasts; 72 hours incubation
An estrogen receptor inhibitor abolishes (nearly) effects of resveratrol or estradiol on MnSOD
2.00
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1.00
DMSO
resveratrol + ICI
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estradiol + ICI
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R
Lung fibroblasts; 72 hours incubation
An ER beta agonist, but not an ER alpha agonist reproduces resveratrol effect on MnSOD
2,3-bis(4-hydroxyphenyl) proprionitrile (DPN)
propylpyrazole triol (PPT)
3.00
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2.00 DMSODPN (beta agonist)
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Our working model
ER Cytosol
MAPKERK1/2
NFBNFB
MnSOD alteredROSmetabolism
Nucleus
MnSOD antiapoptoticMnSOD
Othergenes Mitochondria
Delivering results
Problems with resveratrol bioavailability
Dietary resveratrol delivery in mice
Increasing brain MnSOD
Jonathon Brotchie
Dietary resveratrol delivery in mice
Increasing brain MnSOD
2.000
2.500
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Control M
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Control
Silicon Control
Silicon Res
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Brain
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Silicon nanocapsule delivery of resveratrol with Paul Zelisko
Conclusions
In human cells resveratrol interacts with ERbeta Transcriptional upregulation of MnSOD (progressive and big!) Increased MnSOD makes mitochondria resistant to cyt c releaseIncreased MnSOD makes mitochondria resistant to cyt c release Increased MnSOD alters mitochondrial reactive oxygen metabolism Altered ROS metabolism influences cell replication (signal) Red wine consumption provides low amount of bioavailable resveratrolp p Resveratrol-mediated MnSOD induction assisted by delivering in lipid Maybe promote also using silicon nanocapsules (more work)
Future goals
Refine strategy to increase RES bioavailability (RES supplemented wines?)
Explore related polyphenols & phytoestrogens (-quercetin does not work similarly)
E l f RE b l Explore activities of RES metabolites
Human trials?
This work was done by:
Kurtis Salway, M.Sc. candidate
Melissa Page, PhD candidate
Ellen Robb, PhD candidate
Melanie Brown, M.Sc.
Our research is supported by: