Phytochemicals in Grape

Phytochemicals in Grape

by Santosh, Yafei, Magnet

Introduction

The world’s largest fruit crop, with approximately 65 million metric tons produced annually. (Mazza 1995)Total of 1610 phytochemical compound record (Pezzuto 2008)

Introduction

LuteinLycopeneCatechinResveratrolAnthocyaninZeaxanthinQuercetinMelatonin

Resveratrol

Resveratrol

Resveratrol is a naturally occurring polyphenolic phytoalexin A potent antioxidant (Cadenas and Barja, 1999) Potentially benefits Cancer Cardiovascular disease and other diseases Both in vivo and in vitro (Bradamante and others 2004, Pezzuto 2008, Afaq and others 2003, Sengottuvelan and others 2006, Jang and others 1997)

Cardiovascular

disease

Cardiovascular disease

The cardioprotective effect of resveratrol in humans has yet to be demonstrated

(Bradamante and others 2004)

Cardiovascular disease

In vitro and in vivo studies on animal model It may provide protection by a number of mechanisms Inhibition of (LDL) oxidation Inhibition of platelet aggregationSynthesis of proatherogenic eicosanoids Expression of procoagulant tissue factorInhibition of cell proliferationIncreased vasorelaxation and upregulation of NO synthase

(Bradamante and others 2004)

Inhibition of (LDL) oxidation

During LDL oxidation Polyunsaturated fatty acids are converted to fatty acid hydroperoxidesConjugated double bonds with a characteristic absorbance at 234 nm

(Brito and others 2002)

Inhibition of (LDL) oxidation

The increase in the absorbance intensity is significantly reduced by the pre-incubator of LDL with resveratrol (2, 4 or 6 M) in a concentration dependent manner.

pointing out that resveratrol affords an efficient protection against the ferrylmyoglobin-mediated LDL oxidation.

(Brito and others 2002)

Potential Chemoprevention

Chemoprevention

Mediate positive responses with tumor transplant models for gastric cancers (Zhou and others 2005)Hepatoma (Wu and others 2004) Neuroblastoma (Chen and others 2004)Mammary (Garvin and others 2006)Glioma (Chen and others 2006)Laryngeal (Li and others 2005) However, activity was not observed with 4T1 breast cancer (Bove and others 2002) and leukemia (Gao and others 2002)

Chemoprevention

Chemopreventative activity Inhibits the cyclooxygenase activity of COX-1Induces quinone reductase activityReduces inflammatory

(Jang and others 1997)

Inflammation

(Jang and others 1997)

Resveratrol significantly reduced pedal edema both in the acute phase (3 to 7 hours) and in the chronic phase (24 to 144 hours) in the carrageenan-induced model of inflammation in

rats.

Reveratrol (, 3mg per kilogram of body weight; , 8 mg/kg) phenylbutazone () indomethacin () control group ()

COX-1 and COX-2

COX catalyzes the conversion of arachidonic acid Pro-inflammatory substances such as prostaglandinsStimulated tumor cell growthsuppress immune surveillance Activate carcinogens to forms that damage genetic material

COX-1 and COX-2

(Jang and others 1997)

Indomethacin (

) on COX-1 activity Resveratrol on COX-1 () or COX-2 () activity

Resveratrol inhibit the cyclooxygenase activity of COX-1 and this activity correlates with antitumor promotion.Although its inhibitory activity was less than indomethacin

Skin cancer

Resveratrol also shows potentially prevention of short-term ultraviolet B radiation-mediated damages in hairless mice Reduces cutaneous edamaInhibit reactive oxygen speciesInhibit lipid peroxidation

(Afaq and others 2003)

UVB-mediated skin edema

Bi-fold skin thickness and ear punch weight were evaluated 24 h after UVB irradiation To observe the protective effect of resveratrol on UVB-mediated skin edema

(Afaq and others 2003)

UVB-mediated skin edema

The exposure of the mice to UVB (180

mJ/cm2) resulted in a significant increase in bi-fold skin thickness and ear punch weight Pretreatment of skin with resveratrol was found to result in a significant inhibition in the increases in bi-fold skin thickness and ear punch weight 24 h after UVB irradiation compared to the UVB-alone group

(Afaq and others 2003)

Human Clinical trail

Although animal studies data indicate promising activity of resveratrol human clinical trail and studies data are still limited (Walle and others 2004, Zamora-Ros and others 2006)

Plasma concentration

For oral dose, an

early peak of total radioactivity was reached at about 1 h after the dose and at 6 h after the dose, there was second peak For Intravenous, there was a rapid fall of the plasma concentrations of total radioactivity

Oral 25-mg (110 mol) () Intravenous 0.2-mg (0.8 mol) () 14C-resveratrol doses in human subjects

Recovery of total radioactivity in urine and feces

The

overall recoveries in urine and feces were 70.5 to 97.6% after the oral 53.5 to 91.2% after the intravenous dose

LC/MS of urinary excretion

Resveratrol

(RV) Metabolites (M1–M5)

M1 was a resveratrol monoglucuronideM2 was an isomeric resveratrol monoglucuronideM3 was a dihydroresveratrol monoglucuronide.M4 was a resveratrol monosulfateM5 was dihydroresveratrol sulfate

Resveratrol and metabolite in plasma

Resveratrol

(RV) Major sulfate conjugates M4/M5

10-min samples, unchanged resveratrol with an estimated concentration range of 3.7 to 16.4 ng/ml. Two subjects also demonstrated a major metabolite peak with an estimated concentration of 9 to 13.5 ng/ml30-min samples, the plasma from subjects 1 and 2 had no resveratrol, whereas in subject 3 there was a small amount of resveratrol left, but mostly the sulfate conjugate, M4/M5. samples obtained beyond 30 min, there was no unchanged resveratrol detected in any of the subjects.

Anthocyanin

Anthocyanin

Anthocyanins are a group of abundant and widely consumed flavonoid

constituentsAntioxidant activity (Castaneda-Ovando and others 2009)Disease prevention Cardiovascular illnesses Diabetes Visual dysfunction

Figure 1. General anthocyanins structure

Antioxidant activities

React with free radicalsInterrupt the propagation of new free radical speciesChelate metal ions such as Fe2+ Catalyze lipid oxidation to alter their redox potentialsSignificantly improve certain immune responses

(Hertog and others 1993)

Antioxidant activities

Figure 2. Plots showing the correlation of total anthocyanin content of samples with (a) antiradical activity, (b) reducing power and (c) with hydroxyl free radical scavenging activity

Anthocyanins and cardiovascular disease

Increased consumption of anthocyanins lowers the risk of cardiovascular disease (CVD)May regulate different signaling pathways involved in the development of CVD

(Demrow and others 1995)

Anthocyanins and cardiovascular disease

Figure 3. Representative tracing of the hemodynamic effect of intravenous grape juice

(

Demrow and others 1995)

Anthocyanins and anti-diabetic properties

Consumption of fruits and vegetables decrease the incidence of type-2 diabetesProtect pancreatic β-cells from glucose induced oxidative stress

(Al-Awwadi and others 2005)

Anthocyanins and anti-diabetic properties

Figure 4. Insulin secreted by compounds 3 and 7-9 at 4 and 10 mM glucose concentrations

(Jayaprakasam and others 2005)

Anthocyanins and anti-diabetic properties

Figure 5. Percent inhibition of COX-1 and COX-2 enzymes by Vioxx, Celebrex, aspirin, naproxen, and ibuprofen

(Zhang and others 2004)

Anthocyanins and vision improvement

British Royal Air Force aviators ate bilberry jam to improve their night visionImpaired visual function is a techno-stress syndromecaused by concentrated work on video display terminals in today’s computerised society

Anthocyanins and vision improvement

Table 1. Contrast sensitivity before and after administration in placebo and anthocyanoside group

(Lee and others 2005)

Melatonin in grapes

Structure : Precursor : tryptophan.

Sources of melatonin

Metabolic Pathyway

Four enzymes from Try to MelMelatonin regulates your sleep and wake cycles.Light also affects the amount of melatonin production in your body

Dynamic and cyclic nature of melatonin production

Human blood levels range from 20 picograms per milliliter in the morning to 55 pg/mL at night (a picogram is one-trillionth of a gram). Natural production of melatonin by the human body declines with age.Melatonin are used to treat jet lag or Insomnia(lack of sleep),  (SAD).Melatonin supplement how safe are they? Overdose? Contamination? OTC or requires prescription?

Melatonin’s functions in plants

Photo-periodic reactions in higher plants. Night signalsAntioxidant activity ( protection of germen to oxidative stress).Redox cycling doesn’t occur in melatonin(suicidal antioxidant);stable end productGrowth stimulating factor

Melatonin concentration in plants??

GRAPES

Melatonin in grapes:

It is available in wide concentrations ranging from pg to

ug

/g tissue ( cultivars, ripeness, harvest time , light intensity).

Skins and seeds contain more melatonin than pulp/juice. Red grapes contain higher melatonin than white one.

In wine , fermentation also generates

melatoinin

synthesized by yeasts.

When plants defense activator,

Benzothiadiazole

(BTH) is used to treat grape vine, increase in concentration of melatonin has been observed.  Melatonin is found considerably higher in some cultivars, such as

Nebbiolo

, and which can be raised further by BTH treatment. 

Melatonin isomers also exists in grapes.

Melatonin concentration in the eight grape cultivars examined and in the two plasma samples used as control

SampleMelatonina (pg ml−1)Melatoninb (ng g−1)Plasma control 110.6 ± 3.2NDPlasma control 2134.7 ± 10.3NDBarbera25.5 ± 2.50.633Croatina304.6 ± 23.40.870Cabernet Sauvignon183.6 ± 18.80.422Cabernet Franc2.4 ± 0.60.005Marzemino14.9 ± 4.10.031Nebbiolo428.3 ± 32.10.965Sangiovese255.5 ± 16.30.332Merlot114.2 ± 7.20.264Merlot, BTH-treated301.5 ± 22.50.726aMelatonin concentration in plasma and grape skin extracts as determined by ELISA (means ± SE from three extraction experiments).bMelatonin content in ng, referred to 1 g of grape skin and calculated from the means in the other column of data.Iriti et al.(2006)

HEALTH SIGNIFICANCE

Is melatonin a Panacea ??

References supporting antioxidant activities of melatonin

Galano

, A., Tan, D. X., & Reiter, R. J. (2011). Melatonin as a natural ally

against oxidative stress

: a physicochemical examination. 

Journal of pineal research

, 

51

(1), 1-16.

Rodriguez, C., Mayo, J. C.,

Sainz

, R. M.,

Antolin

, I., Herrera, F.,

Martín

, V., & Reiter, R. J. (2004). Regulation of

antioxidant enzymes

: a significant role for melatonin. 

Journal of pineal research

, 

36

(1), 1-9.

Tan, D. X., Manchester, L. C.,

Terron

, M. P., Flores, L. J., & Reiter, R. J. (2007).

One molecule, many derivatives

: A never‐ending interaction of melatonin with reactive oxygen and nitrogen species?. 

Journal of pineal research

, 

42

(1), 28-42.

Reiter, R. J., Tan, D. X.,

Osuna

, C., &

Gitto

, E. (2000). Actions of melatonin in the

reduction of oxidative stress

. 

Journal of Biomedical Science

, 

7

(6), 444-458.

Antioxidant activity : mechanisms

Figure . Hypothetical pathways involved in melatonin regulation of antioxidant enzyme gene expression and activity.

Some of the proposed mechanisms by which melatonin interferes with the growth of estrogen-mediated mammary cancer cell growth. 

References supporting to treat sleep disorders in Alzheimer’s disease patients.

Cardinali

, D. P.,

Brusco

, L. I.,

Liberczuk

, C., &

Furio

, A. M. (2002). The use of melatonin in Alzheimer's disease. 

Neuro

endocrinology letters

, 

23

, 20-23.

Sewerynek, E. (2002). Melatonin and the cardiovascular system. Neuro endocrinology letters, 23, 79.

Melatonin may reduce blood pressure via the following mechanisms:

1) by a direct effect on the hypothalamus;

2) as an antioxidant which lowers blood pressure;

3) by decreasing the level of

catecholamines

, or

4) by relaxing the smooth muscle in the aorta wall.

Melatonin and immune system

Carrillo-

Vico

, A., Guerrero, J. M.,

Lardone

, P. J., & Reiter, R. J. (2005). A review of the

multiple actions of melatonin on the immune system. 

Endocrine

,

27

(2), 189-200.

Skwarlo-Sonta

, K. (2002

). Melatonin in immunity: comparative aspects.

Neuro

endocrinology letters

, 

23

, 61.

Szczepanik

, M. (2007).

Melatonin and its influence on immune system

. 

Journal of physiology and pharmacology

, 

58

(6), 115-124.

Hypothetical scheme of melatonin regulation of cytokine production.