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Chedea, V.S.;  Macovei, �.O.;  Bocșan, I.C.;  Măgureanu, D.C.;  Levai, A.M.;  Buzoianu, A.D.;  Pop, R.M. Grape Pomace Polyphenols Benefic Actions. Encyclopedia. Available online: (accessed on 07 December 2023).
Chedea VS,  Macovei �O,  Bocșan IC,  Măgureanu DC,  Levai AM,  Buzoianu AD, et al. Grape Pomace Polyphenols Benefic Actions. Encyclopedia. Available at: Accessed December 07, 2023.
Chedea, Veronica Sanda, Ștefan Octavian Macovei, Ioana Corina Bocșan, Dan Claudiu Măgureanu, Antonia Mihaela Levai, Anca Dana Buzoianu, Raluca Maria Pop. "Grape Pomace Polyphenols Benefic Actions" Encyclopedia, (accessed December 07, 2023).
Chedea, V.S.,  Macovei, �.O.,  Bocșan, I.C.,  Măgureanu, D.C.,  Levai, A.M.,  Buzoianu, A.D., & Pop, R.M.(2022, October 19). Grape Pomace Polyphenols Benefic Actions. In Encyclopedia.
Chedea, Veronica Sanda, et al. "Grape Pomace Polyphenols Benefic Actions." Encyclopedia. Web. 19 October, 2022.
Grape Pomace Polyphenols Benefic Actions

Grape pomace polyphenols research studies have grown in the last decades, given their potential benefic effects on promoting human health. Some of their benefic actions are observed in oxidative stress and inflammation aiming at homeostasis restoration. Regarding the antioxidant effect, polyphenols can modulate the endogenous pathway responsible for combating oxidative stress. These effects can be achieved by polyphenols capacity to activate the nuclear factor E2 and to up-regulate superoxide dismutase, catalase, glutathione, glutathione peroxidase, and heme-oxidase 1 or their capacity to scavenge and chelate reactive oxygen species involved in ROS production. In inflammation, polyphenols are reported to inhibit the mitogen-activated kinase pathway, Nf-kB, anddown-regulate cytokines and chemokines. Polyphenols also inhibit cyclooxygenase and lipoxygenase, which are involved in the arachidonic acid signaling pathway, being responsible for synthesizing prostaglandin, thromboxane A2, and leukotrienes which further increase inflammatory response.

grape pomace polyphenols antioxidant anti-inflammatory

1. In Vitro Beneficial Actions of Grape Pomace in Oxidative Stress and Inflammation

The in vitro studies, as presented in Table 1, can offer the possibility to investigate and identify the diversity of related diseases in which GP exerts the optimum antioxidant and anti-inflammatory effects.
Table 1. In vitro beneficial actions of grape pomace in oxidative stress and inflammation.
The in vitro beneficial action of GP was studied by Goutzourelas et al. (2015) [1]. They investigated an extract of red GP on muscle and endothelial cells using non-cytotoxic doses to check the effect of GP polyphenols extract on cells’ antioxidant enzymes [1]. The red GP extract was investigated as a mixture of compounds that contained phenolic acids (caftaric acid, gallic acid), anthocyanins, flavanols (epicatechin and catechin), flavonols (quercetin), and anthocyanidins. It was observed that GP treatment increased Glutathione S-transferase (GST) and GSH levels in both cell lines. CAT levels were decreased in endothelial cells, while in muscle cells it showed no significant differences. SOD and HO-1 presented no differences in any population. An explanation for these inconstant findings, in which some of the antioxidant enzymes are not modified, is the ability of GP to enhance other antioxidant systems (GSC, GSH) [1] (Table 1). Another in vitro study, of Pop et al. (2022), investigated the antioxidant effect of red GP (mixture of Pinot Noir, Cabernet Sauvignon, Fetească Neagră, and Mamaia cultivars) and white GP (mixture of Sauvignon Blanc and Muscat Ottonel cultivars) added to a mouthwash on both H2O2 exposed and non-exposed fibroblast cells [7]. They observed that both red grape pomace (RGP) and white grape pomace (WGP) decreased ROS levels in a dose-dependent matter (100 < 200 < 300 µg/mL). Similar to the non-exposed condition, in the presence of H2O2, red GP and white GP led to a significant decrease in ROS levels, the only difference being that while red GP effect was dose-dependent, and white GP produced a non-dependent action [7]. Moreover, they also studied the anti-inflammatory effects of these extracts on lipopolysaccharides (LPS) induced inflammation in cells [7]. It was observed that while in the case of white GP a dose of 100 µg/mL was sufficient to induce a significant reduction of interleukin (IL) -8 levels, for red GP was necessary a higher dose of 200 µg/mL. At the dose of 300 µg/mL, both extracts significantly reduced IL-8 levels, but not even the highest dose did significantly reduce the levels of IL-6. In the case of IL-1β, the lowest dose, 100 µg/mL, reduced its level to a similar one found in the non-exposed cells, while the doses of 200 and 300 µg/mL reduced, even more, the levels of IL-1β [7].
Marzulli et al. (2018), treated mononuclear cells with phorbol 12-myristate 13-acetate (PMA) to activate inflammation, and with different GPs (red Negroamaro cultivar or white Koshu cultivar) extracts (water, ethanol), to observe their immunomodulatory effects [8]. In terms of cytokine release, all GP fractions and extracts increased anti-inflammatory (IL-10) and pro-inflammatory (IL-12, IL-1β, IL-6, tumor necrosis factor-alpha (TNF-α)) cytokines. The water extracts of both GPs managed to increase T regulatory cells and forkhead box P3 (FoxP3) protein, which is responsible for the genes activity control that are involved in the immune system regulation. Another benefic effect of GPs extracts is FoxP3 increase which is a marker with a role in stabilizing the T regulatory cells’ function. All extracts lowered the release of granzyme (GrB) compared to PMA treated group [8]. GrB is an enzyme secreted by cytolytic T cells with role in cell necrosis leading to harmful effects on homeostasis [8]. Regarding intracellular cytokines, the water extract of red Negroamaro GP increased TNF-α and IL-10 content in monocytes, while the red Negroamaro GP ethanol extract increased IL-12 and IL-10 levels in lymphocytes. Further, the white Koshu GP water extract increased monocyte levels of IL-10 and IL-12, while the white Koshu GP ethanol extract increased lymphocyte levels of TNF-α and IL-10. IL-10 was increased by both water or ethanolic, red or white GP extracts and as underlined by authors [8], the release of IL-10 by T cells and monocytes is a key step in maintaining the immune homeostasis. In conclusion, GPs extracts could induce immune homeostasis through the anti-inflammatory IL-10 secretion which counterbalances the pro-inflammatory cytokines (IL-12 and TNF-α) [8]. Another study that reinforces the anti-inflammatory effects of RGP from Vitis vinifera L. cv. Montepulciano d’Abruzzo on LPS-stimulated macrophages is that of Mollica et al. (2021). They observed that the extract significantly inhibited the release of cytokines (IL-6, TNF-α, and IL-1β), the maximal inhibitory action being at the dose of 100 μg/mL [9].
The possible potential impact of GP extracts on in vitro calcitonin gene-related peptide (CGRP) secretion was investigated as a potential mechanism to influence migraine [2]. The treatment of CA 77 cells with different red GP extracts showed a significant decrease in CGRP levels. CGRP is a gene that represents a key mediator of migraine-induced inflammation [2]. The results suggest that GP extracts had anti-inflammatory effect preventing the release of CGRP in migraine [8885].
White GP extract and a mixture of red and white GP extract, in different concentrations (100, 200, 500 μg/mL dry extract w/v), were added to Caco-2 cells after treatment with an inflammation inducer (IL-1β) to observe the effects on IL-8 secretion and NF-κB expression [10]. Grape pomaces were hydrolyzed enzymatically to determine if anti-inflammatory effects would be augmented. Both white and red GP contained quercetin, catechin, resveratrol, gallic and caffeic acids, trans-resveratrol, rutin, and procyanidin B2 [10]. All GP fractions (100, 200 μg/mL dry extract w/v) with or without enzymatic transformation decreased ROS levels, while treatment with GP extracts in higher concentration (500 μg/mL dry extract w/v) showed a considerable increase in ROS levels. Furthermore, NF-κB expression and prostaglandin E2 (PGE2) levels were significantly reduced in all fractions. At the same time, IL-8 secretion revealed a more substantial drop in enzymatically treated fractions of mixed GP, presenting beneficial effects of enzyme hydrolysis. The mixed GP had a more potent anti-inflammatory effect due to the high content of anthocyanins found in red GP [6].
Concerning the benefic antioxidant and anti-inflammatory GP actions, the literature presents a large variety of experimental settings that can be considered for future in vivo research. Also, it can be observed that there is still space for other hypotheses, for both red and white GPs, but especially for the white ones which were much less investigated.

2. In Vivo Beneficial Actions of Grape Pomace in Oxidative Stress and Inflammation

The effect of GP extracts on the pathophysiology of oxidative stress and inflammation in various types of diseases can be well documented using different in vivo experimental models. These types of studies are very important in deciding whether the GP can be further used in safe conditions in human clinical trials.
The antioxidant and anti-inflammatory effects of both fresh and fermented GP extracts (Vitis vinifera L. cultivars, Fetească neagră, and Pinot noir, from Romania) were investigated using and a rat model of induced inflammation by turpentine oil [11]. The administration of turpentine oil increased the total oxidative status, oxidative stress index and reduced total antioxidant reactivity [11]. Treatment with GP decreased total oxidative status and oxidative stress index in a dose-dependent manner, but total antioxidant reactivity was not modified. All GP’s fractions significantly reduced malondialdehyde (MDA) levels. Total thiols were considerably lessened by turpentine, but GP managed to increase them in a concentration-dependent way. The same results were observed in the case of NOx production. 3NT was also increased by turpentine, but GP varieties decreased the levels. Due to higher phenolic content, the fresh extract showed a higher antioxidant effect. MDA is a lipid peroxidation waste product with hazardous potential for normal homeostasis. Thiols, under oxidative stress, manage to form disulphide bonds between them to reduce oxidative stress. NO presents a dual effect based on its concentrations. Small doses possess an antioxidant effect, while high doses can cause an increase in oxidative stress through the synthesis of new and stronger radicals. 3NT is a waste product resulting from tyrosine nitration induced by reactive nitrogen species [11]. The authors concluded that GP extracts could be used considered a potential agent in nutraceuticals formulation.
An interesting study that evaluates the effects of red GP flour dietary inclusion on growth, anti-inflammatory, antioxidant, innate-adaptive immunity, and on immune genes expression was performed on Labeo rohita fish against Flavobacterium columnaris induced infection [12]. Treatment with 200 and 300 mg GP flour showed a significant increase in GSH, SOD, and GPx activities as compared to regular diet or 100 mg GP supplementation, in both infected and uninfected groups. Regarding GP action on innate-adaptive immune activity, higher doses of GP (200, 300 mg) increased phagocytosis, alternative-complement pathway activity, raised IgM levels, and serum lysozyme (Lyz) activity when compared to regular diet or 100 mg GP supplementation in infected or uninfected group. In terms of immune-related genes, Lyz, (β-2 microglobulin) β-2M, 3rd component complement (CC3), and immunoglobulin M (IgM) gene expression pointed out a significant growth in infected fish with 200, 300 mg GP supplementation compared to other groups. However, the uninfected group treated with the same doses of GP showed higher gene expression than the infected group. Antioxidant related-genes were measured, and SOD, GPx, nuclear factor erythroid 2-related factor 2 (Nrf2), and (natural killer-cell enhancing factor β) NKEF-β were remarkably higher in all groups treated with raised doses of GP compared to 100 mg GP diet or regular diet in infected or uninfected groups. Furthermore, uninfected groups treated with high doses of GP showed a more significant increase in SOD and GPx expression levels. As for pro-inflammatory-related genes, IL-1β and TNF-α were not modified in any group. Hepcidin and toll-like receptor-22 (TLR22) expression were increased in infected and uninfected groups treated with a high dose of GP [12].
Therefore, in Rajković et al. (2022), GP was given to piglets to assess their positive effects on the animal organism without antibiotics side effects [13]. During the experiment tissue samplings (liver, jejunum, ileum) were collected on days 27/28 and 55/56, while blood samples were taken on days 6, days 27/28, and 55/56. Regarding antioxidant enzymes, GPx (GPx1-liver, GPx-2 jejunum, and ileum) wasn’t different between diets, but the enzyme activity was significantly increased on days 55/56 compared to 27/28 [13]. About, SOD and Manganese Superoxide Dismutase (Mn-SOD) enzymes, there weren’t any differences between diets in jejunum, ileum, and liver, but there was an increase between sampling dates, in days 55/56 compared to 27/28 in the liver. The copper superoxide dismutase system (Cu-SOD or SOD1) presented no distinction between any sampling days in the liver or ileum. CAT activity wasn’t affected by any of the diets in the jejunum and liver, but there were differences between sampling days in the liver and ileum. TBARS concentrations weren’t affected by diets in any organs, only in the jejunum between sampling days (decreased levels on days 55/56 compared to 27/28). GPx2 and SOD1 gene expression were modified at the jejunum level (decreased in days 55/56 compared to 27/28), while CAT expression presented the same results at the ileum level. In the liver, the authors have observed differences between samples for SOD1, CAT, and GPx1 in the liver, with a higher expression on days 27/28 compared to 55/56. In terms of inflammation, pig major acute phase-protein serum levels presented a decrease on days 55/56 and 27/28 compared to day 6 [13]. MDA serum levels decreased through sampling days while for SOD different fluctuations were noticed, without showing any significant values on day 55/56 versus other time points. As a speculative explanation for the variation of antioxidant enzymes decreasing it can be stated that the systemic presence of antioxidant substances can lead to a decreasing need for endogenous antioxidant enzymes production [13].
Another important direction in GP research is to check whether it is suitable to be used as an adjuvant treatment in different pathologies to reduce conventional drugs side effects. Thus, in Mossa et al. (2015) study, cypermethrin was given to female rats to observe toxic effects on the liver and kidneys, and white GP was added to check whether it can counter these toxic effects [14]. The assessment of kidneys and liver biomarkers showed a dose-dependent fall in liver enzymes: aspartate transaminase (AST), alanine transaminase (ALT), gamma-glutamyl transferase (GGT), and alkaline phosphatase (ALP), and a decrease kidneys urea nitrogen and creatine. Also, total proteins and albumin revealed a significant increase in GP treated group. The histological analysis pointed out significant changes due to inflammatory infiltrate in cypermethrin groups, while the GP supplemented group had regressed values for all biomarkers. Similar results were also observed in histological studies of kidneys samples. This may be due to the antioxidant effects of the white GP [14]. This study offers important evidence regarding the use of GP extract with hepatorenal protective activity and encourages future studies to investigate whether it can be used to reduce other drugs adverse reactions.
So far, the existing studies on GP suggest that through its anti-inflammatory and antioxidant effects, GP can be considered a potent agent that can contribute to the restoration of homeostasis to control levels or that can reduce different drug side effects (Table 2).
Table 2. In vivo beneficial actions of grape pomace in oxidative stress and inflammation.


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