Subhendu Mukherjee, Jocelyn I Dudley, Dipak K Das

Cardiovascular Research Center, University of Connecticut HealthCenter, School of Medicine, 263 Farmington Avenue, Farmington, CT

06030-1110, USA

Resveratrol, a Polyphenolic Antioxidant, Present in Grape Skin, is Dose Dependent in Providing Health

Benefits

cis - resveratrol

trans - resveratrol

Source Resveratrol concentration

100% Natural peanut butterBilberries Blueberries Boiled peanuts Cranberry raw juice Dry grape skin Grapes Peanut butter Peanuts Pistachios Ports and sherries Ref grape juice Red winesRoasted peanuts White grape juiceWhite wines

~0.65 μg/g~16 ng/g~32 ng/g~5.1 μg/g~0.2 mg/L~24.06 μg/g0.16–3.54 μg/g0.3–1.4 μg/g0.02–1.92 μg/g0.09–1.67 μg/g<0.1 mg/L ~0.50 mg/L0.1–14.3 mg/L~0.055 μg/g~0.05 mg/L<0.1–2.1 mg/L

The French paradox

Infa

rct s

ize

(%)

10

20

30

40

**

*

*

*

Control-IR

White Wine-I

R

Tyrosol-IR

Hydroxytyrosol-IR

Red Wine-I

R

Resvera

trol-IR

Ethanol-IR

Effect of white wine, its components tyrosol and hydroxytyrosol, red wine, and resveratrol on the infarct size.

Dudley et al., J Agric Food Chem. 2008

0

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45 *

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Control White wine Tyrosol Hydroxytyrosol

Red wine ResveratrolI/R

A

C

Car

diom

yocy

te a

popt

osis

(%)

Effect of white wine, its components tyrosol and hydroxytyrosol, red wine, and resveratrol on the infarct size.

Dud

ley

et a

l., J

Agr

ic F

ood

Che

m. 2

008

B

Bcl2Akt

p-Akt

GAPDH

Bax

HO-1

COX-1

COX-2

e-NOS

i-NOS

GAPDH

Control-B

L

White Wine-I

R

Tyrosol-IR

Hydroxytyrosol-IR

Red Wine-I

R

Resvera

trol-IR

Control-IR

Control-B

L

White Wine-I

R

Tyrosol-IR

Hydroxytyrosol-IR

Red Wine-I

R

Resvera

trol-IR

Control-IR

Trx1

Trx2

GAPDH

Control-B

L

White Wine-I

R

Tyrosol-IR

Hydroxytyrosol-IR

Red Wine-I

R

Resvera

trol-IR

Control-IR

Dudley et al., J Agric Food Chem. 2008

RESVERATROL

Antitumor activity, ChemopreventionInhibits NF-κB activation, Proliferation,Causes S-phase arrest, Induces apoptosisof myeloid leukomia cells

Reduces platelet adhesion,Monocyte adhesion(anti-inflammatory response)

Prevents LDL oxidation

Protects lung from DNADamage and apoptosis

Neuroprotection

Reversible inhibition of herpessimplex virus types 1 & 2 replication

Anti-Aging

Cardioprotection

Protects from UVRadiation injury

Prevents prostate andPancreatic, gastricand thyroid cancer

Protects cerebralischemic injury

Inhibited growth ofH. pylori

Resveratrol

COX / LOX

PKC/PKD

ERK 1/2, JNK, p38 MAPK

PI3K/PKB

AMPKSirt1

PBEFeNOSiNOS

Bax/Bcl2

DNA polymerases α and δ

Quinone reductase 2

Aromatase

Adenylyl cyclase

Src

AktNFκB

p53

Ribonucleotidereductase

STAT3

NO

p56lck

FoxOPGE2

TNF

Molecular targets of Resveratrol

Myocardial injury and ageing related diseases

RESVERATROL

Cancer prevention

RESVERATROL

ApoptosisApoptosis

Anti-apoptoticresveratrol

Pro-apoptoticresveratrol? ? ?

Myocardial injury and ageing related diseases Cancer prevention

Low dose of resveratrol was used

High dose of resveratrol was used

Health beneficiary effects of resveratrol are dose dependent or not?

Panel A Panel B

Sudan-IV stained rabbit aortas with atherosclerotic lesions appearing as darkened areas on a white aortic surface. Panel A) Arteries from control rabbits. Panel B) Arteries from resveratrol-treated rabbits.

High dose resveratrol promotes atherosclerosis in case ofhypercholesterolemic rabbits

Wilson et al., Life Sciences. 1996

Effect of High Dose Resveratrol on Proliferation and Apoptosis in Endothelial and Tumer Cell Culture

Effect of resveratrol on the apoptotic and mitotic index of HT-1080 human fibrosarcoma cell culture

Szende et al., Experimental And Molecular Medicine 2000

Effect of resveratrol on the apoptotic and mitotic index of HT-29 cell culture

Effect of resveratrol on the apoptotic and mitotic index of HUVEC Endothelial cell culture

-2

-1

0

1

2

3

4

5

6

7

8

0

0.5

1

1.5

2

2.5

3

3.5

Control 100 μg/ml 10 μg/ml 1 μg/ml

Control 100 μg/ml 10 μg/ml 1 μg/ml 0.1 μg/ml

Effect of resveratrol on theproliferation of HUVEChuman endothelial cells

Resveratrol

Effect of Reserveratol on the proliferation of HT-29human colon carcinoma cells

Resveratrol

24h treatment48h treatment

Szende et al., Experimental And Molecular Medicine 2000

Resveratrol Associated Renal Toxicity is Dose Dependent

Crowell et al., Toxicological Sciences. 2004

Influence of resveratrol (RSV) concentration on the viability of HEK 293 cells before and after UV irradiation

Caddeo et al., International Journal of Pharmaceutics. 2008

Dumazet et al. (2002) showed that at higher dose resveratrol inhibits the growth and induces apoptosis in case of both normal and leukemic hematopoietic cells

Signorelli et al., (2005) showed that in androgen – sensitive prostatecancer cells, resveratrol had a proliferative activity at a low dose(5 µM), whereas it had a pro-apoptotic activity at a high dose (15 µM or higher)

Zhou et al., (2003) showed that in human esophageal carcinoma cells,resveratrol induces apoptosis when used in high concentration(100 mM) and this high dose of resveratrol also downregulatedBcl2 protein expression and upregulated Bax protein expression

Jang et al., (2006) showed that low concentration (5µM), resveratrolappears to increase cell proliferation, whereas apoptosis is induced invarious cancer cells at 15 µM or higher concentration.

Kyungmin et al. (2006) showed that 100 µM resveratrol induced apoptosis by cleavage of caspase 3 and resveratrol has an inhibitoryeffect on cell migration.

Howitz et al. (2003) showed that the photoprotective effect of resveratrolfrom radiation induced apoptosis in HEK 293 cells was reversed at concentrations greater than 50 µM.

Dose-response curve of the effects of resveratrol on myocardial performance

Control3.7 μM

7.4 μM10 μM

25 μM

Das et al., J Pharmacol Exp Ther. 2006

Dose-response curve of the effects of resveratrol on myocardial performance

Control3.7 μM

7.4 μM10 μM

25 μM

Das et al., J Pharmacol Exp Ther. 2006

Con

trol

3.7 μM

7.4 μM

10 μ

M

25 μ

M

Con

trol

3.7 μM

7.4 μM

10 μ

M

25 μ

M

Dose-response curve of the effects of resveratrol onmyocardial infarction and cardiomyocyte apoptosis.

Das et al., J Pharmacol Exp Ther. 2006

2 h 45 min Perfusion with KHB

15’

15’

30’ Ischemia 2 h Reperfusion

30’ Ischemia 2 h Reperfusion

I/R

Infarct size Apoptosis Biochemical and Molecular Biological Studies

Control

Resveratrol Treated

Isolated Working Heart Model of Ischemia-Reperfusion

Dudley et al., Journal of Nutritional Biochemistry. 2008

Dudley et al., Journal of Nutritional Biochemistry. 2008

Dudley et al., Journal of Nutritional Biochemistry. 2008

0

10

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50

Myo

card

ial i

nfar

ct s

ize

(%)

Con

trol I

/R

2.5m

g/kg

5 m

g/ k

g

25 m

g/kg

50 m

g/kg

* *

*

Resveratrol

Dudley et al., Journal of Nutritional Biochemistry. 2008

Effects of high and low doses of resveratrol on the myocardial infarct size

0

5

10

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30C

ardi

omyo

cyte

apo

ptos

is(%

)

Con

trol I

/R

2.5m

g/kg

5 m

g/ k

g

25 m

g/kg

50 m

g/kg

* *

*

Resveratrol

Dudley et al., Journal of Nutritional Biochemistry. 2008

Effects of high and low doses of resveratrol on the myocardial infarct size

TRX-2

TRX-1

GRX-1

GRX2

Cont

rol

I/R 2.5

mg/

kg

5mg/

kg

25m

g/kg

50m

g/kg

GAPDH

Resveratrol

Dudley et al., Journal of Nutritional Biochemistry. 2008

Effects of high and low doses of resveratrol on mRNA transcript of some redox genes

0

0.2

0.4

0.6

0.8

1

0.2

0.4

0.6

0.8

1

Control I/R 2.5mg/kg 5mg/kg 25mg/kg 50mg/kg

Rel

ativ

e ba

nd in

tens

ity(A

rbitr

ary

units

)

Resveratrol

Ref 1

GAPDH

Contro

l

I/R2.5

mg/kg

5mg/k

g25

mg/kg

50m

g/kg

Resveratrol

Dudley et al., Journal of Nutritional Biochemistry. 2008

Effects of high and low doses of resveratrol on Ref1 protein induction

Control I/R 2.5mg/kg 5mg/kg 25mg/kg 50mg/kg0

0.5

1

1.5

2

2.5

3p-AktBcl 2

*

* †

* † * †

*

***

†

Rel

ativ

e ba

nd in

tens

ity(A

rbitr

ary

units

)

GAPDH

Bcl 2Akt

P-Akt

Contro

l

I/R2.5

mg/kg

5mg/k

g25

mg/kg

50m

g/kg

Resveratrol

ResveratrolDudley et al., Journal of Nutritional Biochemistry. 2008

Effects of high and low doses of resveratrol on survival signal

•Resveratrol is good for health but the health benefit of resveratrol is dose dependent.

•Low doses resveratrol protects from different types of diseases such as cardiovascular, ageing etc.

• High doses resveratrol can be detrimental for normal tissue but it can be used in cancer prevention.