Phytochemicals in Grape by Santosh Yafei Magnet Introduction
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 • • Lutein Lycopene Catechin Resveratrol Anthocyanin Zeaxanthin Quercetin Melatonin
Resveratrol
Resveratrol § Resveratrol is a naturally occurring polyphenolic phytoalexin § A potent antioxidant § Potentially benefits (Cadenas and Barja, 1999) • 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 aggregation Synthesis of proatherogenic eicosanoids Expression of procoagulant tissue factor Inhibition of cell proliferation Increased 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 hydroperoxides § Conjugated 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 • Hepatoma • Neuroblastoma • Mammary • Glioma • Laryngeal • However, activity was not observed with 4 T 1 breast cancer and leukemia (Zhou and others 2005) (Wu and others 2004) (Chen and others 2004) (Garvin and others 2006) (Chen and others 2006) (Li and others 2005) (Bove and others 2002) (Gao and others 2002)
Chemoprevention • Chemopreventative activity – Inhibits the cyclooxygenase activity of COX-1 – Induces quinone reductase activity – Reduces inflammatory (Jang and others 1997)
Inflammation Reveratrol (� , 3 mg per kilogram of body weight; � , 8 mg/kg) phenylbutazone (� ) indomethacin (� ) control group (� ) 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. (Jang and others 1997)
COX-1 and COX-2 • COX catalyzes the conversion of arachidonic acid – Pro-inflammatory substances such as prostaglandins – Stimulated tumor cell growth – suppress immune surveillance • Activate carcinogens to forms that damage genetic material
COX-1 and COX-2 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 (Jang and others 1997)
Skin cancer • Resveratrol also shows potentially prevention of short-term ultraviolet B radiation-mediated damages in hairless mice § Reduces cutaneous edama § Inhibit reactive oxygen species § Inhibit 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 m. J/cm 2) 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 Oral 25 -mg (110 mol) (� ) Intravenous 0. 2 -mg (0. 8 mol) (� ) 14 C-resveratrol doses in human subjects • 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
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 (M 1–M 5) • • • M 1 was a resveratrol monoglucuronide M 2 was an isomeric resveratrol monoglucuronide M 3 was a dihydroresveratrol monoglucuronide. M 4 was a resveratrol monosulfate M 5 was dihydroresveratrol sulfate
Resveratrol and metabolite in plasma • Resveratrol (RV) • Major sulfate conjugates M 4/M 5 • 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/ml • 30 -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, M 4/M 5. • 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 constituents § Antioxidant activity § Disease prevention (Castaneda-Ovando and others 2009) • Cardiovascular illnesses • Diabetes • Visual dysfunction Figure 1. General anthocyanins structure
Antioxidant activities § React with free radicals • Interrupt the propagation of new free radical species § Chelate metal ions such as Fe 2+ • Catalyze lipid oxidation to alter their redox potentials § Significantly 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 diabetes § Protect 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 m. M 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 vision § Impaired visual function is a technostress syndrome • caused 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 n Structure : Precursor : tryptophan.
Sources of melatonin
Metabolic Pathyway n Four enzymes from Try to Mel n Melatonin regulates your sleep and wake cycles. n Light also affects the amount of melatonin production in your body
Dynamic and cyclic nature of melatonin production n Human blood levels range from 20 picograms per milliliter in the morning to 55 pg/m. L at night (a picogram is one-trillionth of a gram). n Natural production of melatonin by the human body declines with age. n Melatonin are used to treat jet lag or Insomnia(lack of sleep), (SAD). n Melatonin supplement how safe are they? Overdose? Contamination? OTC or requires prescription?
Melatonin’s functions in plants n Photo-periodic reactions in higher plants. n Night signals n Antioxidant activity ( protection of germen to oxidative stress). Redox cycling doesn’t occur in melatonin(suicidal antioxidant); stable end product n Growth stimulating factor
Melatonin concentration in plants? ?
GRAPES Melatonin in grapes: n It is available in wide concentrations ranging from pg to ug/g tissue ( cultivars, ripeness, harvest time , light intensity). n Skins and seeds contain more melatonin than pulp/juice. Red grapes contain higher melatonin than white one. n In wine , fermentation also generates melatoinin synthesized by yeasts. n 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. n Melatonin isomers also exists in grapes.
Melatonin concentration in the eight grape cultivars examined and in the two plasma samples used as control Melatonina (pg ml− 1) Melatoninb (ng g− 1) Plasma control 1 10. 6 ± 3. 2 ND Plasma control 2 134. 7 ± 10. 3 ND Barbera 25. 5 ± 2. 5 0. 633 Croatina 304. 6 ± 23. 4 0. 870 Cabernet Sauvignon 183. 6 ± 18. 8 0. 422 Cabernet Franc 2. 4 ± 0. 6 0. 005 Marzemino 14. 9 ± 4. 1 0. 031 Nebbiolo 428. 3 ± 32. 1 0. 965 Sangiovese 255. 5 ± 16. 3 0. 332 Merlot 114. 2 ± 7. 2 0. 264 Merlot, BTH-treated 301. 5 ± 22. 5 0. 726 Sample a Melatonin concentration in plasma and grape skin extracts as determined by ELISA (means ± SE from three extraction experiments). b Melatonin 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 n Is melatonin a Panacea ? ?
References supporting antioxidant activities of melatonin n 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. n 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. n 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. n 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 n 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. n 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. n n n 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 n 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. n Skwarlo-Sonta, K. (2002). Melatonin in immunity: comparative aspects. Neuro endocrinology letters, 23, 61. n 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.
- Slides: 51