biological activities of resveratrol

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Biological activities of resveratrol 1 The research program: cell and molecular biology of longevity 1. The research program: cell and molecular biology of longevity 2. Biological activities of resveratrol in mammals (including humans)

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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

    e

    a

    r

    s

    )

    100

    m

    L

    i

    f

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    s

    p

    a

    n

    10

    M

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    u

    m

    L

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    g

    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

    v

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    2.5

    *

    myoblasts Neuroblastoma cells

    3.5

    ***

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    0

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    1.5

    R

    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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    ]

    C2C12180

    200

    *

    P

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    MRC5

    MRC5C2C12

    C2C12

    120

    140

    160

    180

    H

    y

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    SHSY5Y

    SHSY5YMRC5

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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

    *C2C1244.5

    **SHSY5YMRC5

    MRC5C2C12

    2.5

    3

    3.5

    q

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    MRC5SHSY5Y

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    ResveratrolControl0

    0.5

    1

    ResveratrolControl

    Figure 1: Chronicexposureofmusclecells,lungcellsandneuralcellsto50Mresveratrolaffordsprotectionagainstcelldeathinducedbytheoxidativestressorshydrogenperoxideandparaquat.DatapointrepresentsLD50foragiventoxin,Cellsurvivalwasevaluatedbytrypanblueexclusion.Datarepresentsthemeanof5independentmeasurements SEM.*=P

  • Greater MnSOD levels paralleled by reduced replicative growth rate

    50

    Human lung fibroblast cells

    35404550

    i

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    g

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    15202530

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    ControlRES

    05

    1015

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    0 50 100 150 200

    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

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    4.5

    l

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    2

    2.5

    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

    M

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    estradiol + ICI

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    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

    l

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    2.00 DMSODPN (beta agonist)

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    1.00PPT (alpha 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

    D

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    1.500

    2.000

    Control M

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    1.000

    Control

    Silicon Control

    Silicon Res

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    0.500

    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: