The Immunomodulatory and Anti-Inflammatory Role of Polyphenols

This review offers a systematic understanding about how polyphenols target multiple inflammatory components and lead to anti-inflammatory mechanisms. It provides a clear understanding of the molecular mechanisms of action of phenolic compounds. Polyphenols regulate immunity by interfering with immune cell regulation, proinflammatory cytokines’ synthesis, and gene expression. They inactivate NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) and modulate mitogen-activated protein Kinase (MAPk) and arachidonic acids pathways. Polyphenolic compounds inhibit phosphatidylinositide 3-kinases/protein kinase B (PI3K/AkT), inhibitor of kappa kinase/c-Jun amino-terminal kinases (IKK/JNK), mammalian target of rapamycin complex 1 (mTORC1) which is a protein complex that controls protein synthesis, and JAK/STAT. They can suppress toll-like receptor (TLR) and pro-inflammatory genes’ expression. Their antioxidant activity and ability to inhibit enzymes involved in the production of eicosanoids contribute as well to their anti-inflammation properties. They inhibit certain enzymes involved in reactive oxygen species ROS production like xanthine oxidase and NADPH oxidase (NOX) while they upregulate other endogenous antioxidant enzymes like superoxide dismutase (SOD), catalase, and glutathione (GSH) peroxidase (Px). Furthermore, they inhibit phospholipase A2 (PLA2), cyclooxygenase (COX) and lipoxygenase (LOX) leading to a reduction in the production of prostaglandins (PGs) and leukotrienes (LTs) and inflammation antagonism. The effects of these biologically active compounds on the immune system are associated with extended health benefits for different chronic inflammatory diseases. Studies of plant extracts and compounds show that polyphenols can play a beneficial role in the prevention and the progress of chronic diseases related to inflammation such as diabetes, obesity, neurodegeneration, cancers, and cardiovascular diseases, among other conditions.

The immune modulation effect of polyphenols is supported by different studies: some polyphenols impact on immune cells populations, modulate cytokines production, and pro-inflammatory genes expression [ 24 , 25 ]. For example, cardioprotective effects of resveratrol present in red wine grape and nuts were mainly attributed to its anti-inflammatory properties. In vivo and in vitro studies demonstrate that resveratrol can inhibit COX, inactivate peroxisome proliferator-activated receptor gamma (PPARγ) and induce eNOS (endothelial nitric oxide synthase) in murine and rat macrophages [ 26 , 27 , 28 ]. Likewise, a resveratrol analog, RVSA40, inhibits the pro-inflammatory cytokines TNF-α (Tumor necrosis factor alpha) and IL-6 (interleukin-6) in RAW (Murine macrophages cell line) 264.7 macrophages [ 29 ]. Another example is the non-flavonoid curcumin found in turmeric plants and mustard. Curcumin was shown to reduce the expression of inflammatory cytokines: TNF and IL-1, adhesion molecules like ICAM-1 (intercellular adhesion molecule-1) and VCAM-1 (vascular cell adhesion molecule-1) in human umbilical vein endothelial cells and inflammatory mediators like prostaglandins and leukotriens. It also inhibits certain enzymes involved in inflammation like COX in mice (cyclooxygenase), LOX (lipoxygenase) in, human endothelial cells MAPK (mitogen-activated protein Kinase), and IKK (inhibitor of kappa kinase). Moreover, curcumin downregulates NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) and STAT3 (signal transducer and activator of transcription) and reduces the expression of TLR-2 (toll-like receptor-2) and 4 while, in vivo, it upregulates PPARγ (Peroxisome proliferator-activated receptor gamma) in male adult rats [ 30 , 31 , 32 , 33 , 34 , 35 ]. Caffeic acid phenethyl ester suppresses TLR4 activation and LPS-mediated NF-κB in macrophages, Quercetin was also shown to inhibit leukotriens biosynthesis in human polymorphonuclear leukocytes [ 36 , 37 ]. COX2 expression is also attenuated by ECGC (Epigallocatechin gallate) in colon cancer cell and androgen-independent PC-3 cells of human prostate cancer, gingerol in and piceatannol (EGCG analog found in Norway spruces) leading to NFκ B inactivation [ 30 , 38 , 39 , 40 ]. Furthermore, polyphenols, such as Gingerol and Quercetin can activate the production of adiponectin known for its anti-inflammatory effects [ 30 , 39 ]. Similarily, EGCG blocks NFκ B activation in human epithelial cells and downregulates the expression of iNOS (inducible nitric oxide synthase), NO (nitric oxide) production in macrophages resulting in its immunomodulation [ 38 , 40 , 41 ]. A series of in vitro studies found that other polyphenols like oleanolic acid, curcumin, kaempferol-3-O-sophoroside, EGCG and lycopene inhibit high mobility group box1 protein, an important chromatin protein that interacts with nucleosomes, transcription factors, and histones regulating transcription and playing a key role in inflammation [ 35 ]. All of these examples support the anti-inflammatory effects of polyphenols. Polyphenols’ use is associated with a direct change in the count and differentiation of specific immune cells. An increase in T helper 1(Th1), natural killer (NK), macrophages and dendritic cells (DCs) in Peyer’s patches and spleen is associated with oral administration of polyphenols extracted from the fruit date in male C3H/HeN mice [ 24 ]. In humans, the count of regulatory T cells (Treg or suppressor T cells) characterized by the (CD4 + CD25 + Foxp3+) phenotype and involved in immune tolerance and autoimmune control can be boosted by polyphenols [ 42 , 43 , 44 ]. In vivo, Epigallocatechin-3-gallate, found in green tea and injected to Laboratory inbred strain (BALB)/c mice, rises the number of functional Treg in spleens, pancreatic lymph nodes, and mesentheric lymph nodes [ 45 ]. Similarly, in vitro treatment of Jurkat T cells with EGCG or green tea upsurges the expression of Foxp3 and IL10. Baicalin, a flavone, extracted from Huangqin herb, induces Foxp3 expression in HEK 293 T cells and triggers functional Treg from splenic CD4 + CD25− T cells [ 46 ]. Additionally, flavonoids show an agonistic effect of aryl hydrocarbon receptor (AhR) and bind xenobiotic-responsive elements in promoter regions of certain genes, including Foxp3 rising its expression [ 47 ]. Th1 and Th17 populations are also affected by polyphenols: EGCG reduces the differentiation of Th1 and reduces the numbers of Th17 and Th9 cells in specific pathogen-free C57/BL6 female mice [ 48 ]. Other polyphenols like Baicalin show a reduction of Th17 differentiation in vitro and a diminution of IL-17 expression [ 49 ]. Macrophages are affected by polyphenols as well. Macrophages are known to be a key player in the inflammatory response. They initiate inflammation by secreting pro-inflammatory mediators and cytokines like IL-6 and TNF-α [ 50 ]. Polyphenols repress macrophages by inhibiting cyclooxygenase-2 (COX-2), inducibl

4.1. NFκ B Signaling Pathway NF-κB or nuclear factor kappa-light-chain-enhancer of activated B cells is a complex protein that plays a key role in deoxyribonucleic acid (DNA) transcription, cytokine production and cell survival. It controls immune, inflammation, stress, proliferation and apoptotic responses of a cell to multiple stimuli [ 58 ]. The expression of a large number of genes involved in inflammation is controlled by NF-κB such as COX-2, VEGF (vascular endothelial growth Factor), pro-inflammatory cytokines (IL-1, IL-2, IL-6, and TNFα), chemokines (e.g., IL-8, MIP-1α, and MCP-1), adhesion molecules, immuno-receptors, growth factors, and other agents involved in proliferation and invasion [ 71 ]. NFκ B is located in the cytoplasm, it exists as an inactive non-DNA-binding form. Iκ B proteins (Iκ Bs), are inhibitors proteins that are associated with NFκ B resulting in its inactivation. Iκ Bs include Iκ Bα, Iκ Bβ, Iκ Bγ, Iκ Bε, Bcl-3, precursors p100 and p105 [ 72 ]. Under stimulatory conditions, Iκ B kinase (IKK) phosphorylate IκB proteins leading to successive ubiquitination, consequent degradation of the inhibitory proteins and release of NFκ B dimer. This later can translocate into the nucleus and prompts the expression of particular genes [ 72 ]. Different mechanisms regulate NFκ B activity as per the accumulation and degradation of Iκ B, the phosphorylation of NFκ B, the hyper-phosphorylation of IKK, and the processing of NFκ B precursors [ 73 , 74 , 75 ]. Thus, the inhibition of NFκB can be of a great benefit in controlling inflammatory conditions [ 76 ]. Several polyphenols modulate NFκ B activation and reduce inflammation [ 77 , 78 ]. Quercetin blocks the nuclear translocation of p50 and p65 subunits of NFκ B and represses the expression of pro-inflammatory associated genes, NOS and COX-2 in RAW264.7 macrophages [ 79 ]. It inhibits the phosphorylation of Iκ Bα protein both in vitro (using macrophages) and in vivo (using dextran sulfate sodium (DSS) rat colitis model) leading to inactivation of the NFκ B pathway [ 80 ]. In human mast cells, quercetin prevents the degradation of Iκ Bα, as well as the nuclear translocation of p65 resulting in reduction of TNFα, IL-1β, IL-6 and IL-8 [ 63 ]. It can modulate chromatin remodeling, for example it blocks the recruitment of a histone acetyl transferase called CBP/p300 to the promoters of interferon-inducible protein 10 (IP-10) and macrophage inflammatory protein-2 (MIP-2) genes in primary murine small intestinal epithelial cell. As a result, it inhibits the expression of these pro-inflammatory cytokines [ 81 ]. Quercetin can block the activation of IKK, NFκ B, and it reduces the ability of NFκ B to bind DNA in microglia treated by LPS and IFN-γ in mouse BV-2 microglia [ 82 ]. Luteolin, too, blocks NFκ B activation and inhibits pro-inflammatory genes expression and the cytokines production in murine macrophages RAW 264.7 and mouse alveolar macrophages; it also inhibits IKKs in LPS-induced epithelial and dendritic cells [ 83 ]. In addition, in co-cultured intestinal epithelial Caco-2 and macrophage RAW 264.7 cells, luteolin represses NF-ĸ B activation and TNF-α secretion [ 84 ]. Likewise, Genistein represses LPS-induced activation of NF-ĸ B in monocytes and reduces the inflammation by inhibiting NF-ĸ B activation upon adenosine monophosphate activated protein kinase stimulation in LPS-stimulated macrophages RAW 264.7 [ 83 , 85 ]. Galangin, as well, stops the degradation of Iĸ Bα and the translocation of p65 NF-ĸ B, repressing the expression of TNF-α, IL-6, IL-1β, and IL-8 in mast cell [ 86 ]. EGCG counteracts the activation of IKK and the degradation of Iκ Bα and inhibits NFκ B in culture respiratory epithelial cells and in vivo in male Wistar rats [ 87 , 88 ]. Furthermore, EGCG blocks DNA binding of NFκ B which reduces the expression of IL-12p40 and iNOS in murine peritoneal macrophages [ 89 , 90 ]. Catechin and epichatechin reduce NFκ B activity in PMA-induced Jurkat T cells. Flavonoids can modulate NFκ B activation cascade at early phases by affecting IKK activation and regulation of oxidant levels or at late phases by affecting binding of NF-κ B to DNA in jurkat Tcells [ 91 ]. Hydroxytyrosol, and resveratrol inhibit NFκ B activation, and the expression of VCAM-1 in LPS-stimulated human umbilical vein endothelial cells [ 92 ]. In summary, polyphenols can modulate NFκ B activation cascade at different steps such as by affecting IKK activation and regulating of the oxidant levels or by affecting binding of NF-κ B to DNA leading to an important anti-inflammatory effect responsible for their potential value in treating chronic inflammatory conditions ( Figure 1 ).

Arachidonic acid (AA) is liberated by membrane phospholipids upon phospholipase A2 (PLA2) cleavage. Cyclooxygenase (COX) or lipoxygenase (LOX) metabolize it and produce, respectively, prostaglandins (PGs) and thromboxane A2 (TXA2) by COX, and hydroxyeicosatetraenoic acids and leukotrienes (LTs) by LOX [ 106 ]. The COX family involves different members (COX1, COX-2, and COX-3). COX-2 is responsible of the production of important quantity of prostaglandins, its expression is triggered by lipopolysaccharide and pro-inflammatory cytokines [ 107 ]. The ability of polyphenols to reduce the release of arachidonic acid, prostaglandins, and leukotrienes is considered one of their most important anti-inflammatory mechanisms ( Figure 1 ). Their action is mainly realized by their ability to inhibit cellular enzymes, such as PLA2, COX, and LOX [ 21 , 108 , 109 , 110 , 111 ]. Quercetin blocks COX and LOX in various cell types such as rat peritoneal leukocyte, murine leukocytes, and guinea pig epidermis [ 110 , 112 , 113 ]. Similarly, red wine reduces COX-2 expression in old male F344 rats [ 114 ]. In LPS activated murine macrophages, green tea polyphenols not only suppress NF-κB and MAPK pathways but also constrain the expression of COX-2 and the release of prostaglandin (PGE2) in RAW 264.7 macrophages [ 115 , 116 ]. Equally, a reduction in the release of PGE2 is observed with other polyphenols, such as kaempferol in culture of LPS-stimulated human whole blood cells [ 117 ]. Extra virgin olive oil rich with more than 30 phenolic compounds inhibit 5-LOX in human activated leukocytes reducing leukotriene B4 and suppresses eicosanoids production by animal and human cells in vitro [ 118 , 119 ]. Finally, certain polyphenols show structural and functional similarities with specific anti-inflammatory drugs. A phenolic compound—oleocanthal—demonstrates a natural anti-inflammatory property and exhibits structural similarities to the ibuprofen (a well-known anti-inflammatory drug). Oleocanthal—like ibuprofen—inhibits COX-1 and COX-2 activities in a dose-dependent manner [ 120 ].

Referring to the previously cited roles of polyphenols in maintaining tissue homeostasis by targeting different signaling pathways and referring to their antioxidant, anti-inflammation, and protection against pro-inflammation properties; polyphenols play a beneficial role in the prevention and the process of chronic diseases related to inflammation. Various polyphenolic compounds show protective actions in diabetes, obesity, neurodegeneration, cancers, and cardiovascular diseases, among other conditions [ 30 , 146 , 147 , 148 , 149 , 150 , 151 , 152 , 153 , 154 ]. 6.1. Polyphenols and Insulin Resistance Polyphenols reduce insulin resistance. They promote glycolysis by activation of AMPK (AMP- activated protein kinase) or inhibition of mTORC1 and PI3K/AkT in vivo (in rats), ex vivo (in rats’ muscles strips) and in vitro (in C2C12 myoblasts and HELA cells) [ 148 , 149 , 155 , 156 ]. Additionally, AMPK activation by polyphenols increases glucose uptake by positively affecting eNOS imitating muscle contraction and in vivo activity of insulin [ 148 , 149 , 150 ]. Similarly, it is found that polyphenols lower insulin resistance by inhibiting PI3K/AkT and JNK of activation of the AMPK-SirT1-PGC1α axis (i.e., gingerol and anthocyans, and their ability to protect from diabetes and reduce insulin resistance using in vivo, ex vivo and in vitro studies [ 26 , 27 , 28 , 148 , 149 , 155 ]. In addition, polyphenols attenuate glucose intake from carbohydrates by inhibiting rats’ α-glucosidase [ 157 ]. Lastly, polyphenols, like falvonoids, can improve insulin secretion by reducing apoptosis of pancreatic β–cells [ 145 ].

Polyphenols show protective effects in neurological disease [ 152 , 153 ]. High flavonoid intake can reduce by 50% dementia and aging. More precisely, it lowers the incidence of Parkinson’s and delays the onset of Alzheimer’s disease as per different epidemiological studies [ 167 , 168 , 169 , 170 ]. EGCG has neuroprotective properties due to its antioxidant (SOD, GSHPx) activities and cellular GSH contents and ability to reduce ROS contents. Similarly, anthocyanins neuroprotective characteristics are related to the improvement of oxidative stress and reduction of Aβ deposition [ 38 , 171 , 172 ]. Other mechanisms of polyphenols protection in neurodegenerative diseases is modulation of neuronal and glial signaling pathways [ 173 ]. Polyphenols can downregulate NF-κB related with iNOS generation in glial cells [ 174 , 175 , 176 ]. Moreover, their ability to inhibit monoamine oxidase plays a positive role in cognition, depression, and learning ability in vivo in male laca mice [ 172 ].

Clinical and epidemiological studies have reported that polyphenols have chemo-preventive and anticancer efficacy [ 182 , 183 , 184 ]. Polyphenol compounds have the ability to inhibit the proliferation of different types of cancer such as prostate, bladder, lung, gastrointestinal, breast, and ovarian cancers [ 154 ]. For instance, quercetin, resveratrol, green tea polyphenols [ 185 ], epigallocatechin-3-gallate [ 186 ], and curcumin [ 187 ] have demonstrated efficacy as anticancer compounds. Several studies reported that polyphenols are able to prevent cancer initiation (cyto-protective), progression, recurrence, and metastasis to distant organs (cytotoxic) as per different epidemiological, in vitro, and in vivo studie [ 188 , 189 , 190 ]. However, a dichotomy exists between polyphenols’ antioxidant effects in normal cells, and their potential pro-oxidant effects in cancer cells [ 154 , 188 ]. Recent studies illustrated a direct correlation between ROS in intracellular signaling cascade and carcinogenesis [ 191 ]. Oxidative stress targets proteins, lipids, and DNA/RNA causing changes that increase the risks of mutagenesis. ROS/RNS (reactive nitrogen species) overproduction over a prolonged period of time damages cellular structure and functions and causes somatic mutations such as pre-neoplastic and neoplastic transformations that may lead to cell death by necrotic and apoptotic processes [ 192 ]. Polyphenols compounds contain hydroxyl groups that donate their protons to reactive oxygen species (ROS) [ 193 ]. Moreover, they reduce the activity of phase I enzymes, primarily cytochrome P450 enzymes (CYPs), such as CYP1A1 and CYP1B1 which lead to prevent the formation of reactive and carcinogenic metabolites in human bronchial epithelial cells [ 194 ]. They also can induce phase II enzymes that initiate the formation of polar metabolites which are readily excreted from the body [ 195 ]. Certain dietary polyphenols such as flavonoids reduce cellular formation of ROS which protects from the oxidation of DNA [ 193 ]. In addition to their anti-oxidant properties, pro-oxidant characteristic of polyphenols is important in treating and preventing cancer. Pro-oxidant activity can be initiated by certain conditions such as superoxide leakage [ 196 ]. The pro-oxidant activities of polyphenols in cancer cells can result in inducing apoptosis [ 197 ], cell cycle arrest [ 198 ] and inhibiting the proliferation signaling pathways (i.e., epidermal growth factor receptor/mitogen activated protein kinase, phosphatidylinositide 3-kinases/protein kinase B, as well as NF-ĸB) [ 199 ]. For example, polyphenols from apple are able to inhibit the proliferation of human bladder transitional cell carcinoma (TCC, TSGH-8301 cells), inducing G2/M cell cycle arrest, and promoting apoptosis [ 200 ]. In human papilloma virus-18-positive HeLa cervical cancer cells, green tea polyphenols can induce cell cycle arrest at the subG1 phase and apoptosis through caspases activation [ 201 ]. Flavonoids, such as quercetin, induce apoptosis in many cancer cells such as leukemic U937 cell [ 202 ], prostate cancer cells [ 203 ], hepatic cancer cells [ 204 ], among other types. A combination of quercetin with resveratrol and catechin inhibits breast cancer progression in vitro and in vivo by inducing apoptosis in carcinogenic breast cells [ 205 ]. In addition, polyphenols can reduce cancer metastasis such as quercetin [ 206 , 207 ]. Sufficient studies have reported that NF-κB signaling pathways are closely related to cancer metastasis. Polyphenols can disrupt the metastatic potential of cancer by inhibiting NF-κB activity [ 208 ]. Curcumin is a good example [ 209 , 210 , 211 ] of decreasing cancer metastasis in mice by suppressing NF-κB expression and down-regulating VEGF (vascular endothelial growth factor), COX-2, and MMP-9 (matrix metallopeptidase-9) expression in tissues of the breast, brain, lung, liver, and spleen [ 212 , 213 ]. Moreover, the strength of metastasis is associated to the epithelial-to-mesenchymal transition (EMT) [ 214 ]. There is robust evidence that polyphenols compounds can modulate EMT and its related signaling pathways [ 215 ]. For example, EGCG, a flavan-3-ol, induces apoptosis and significantly reduces colony formation and cell migration in nasopharyngeal carcinoma (NPC) and cancer stem cells (CSC) in different cell lines [ 216 ]. Luteolin and quercetin reverse the migration and invasiveness of metastatic cells by reducing the expression of mesenchymal markers and transcriptional factors on the cell membrane (i.e., twist, snail, and N-cadherin) and upregulating adhesion molecules such as E-cadherin [ 217 ]. Thus, through variable mechanisms, polyphenols broadly downregulate inflammation origination, progression, and evolution to cancers ( Figure 3 ). In order to emphasize on the beneficial health effects of polyphenols, different medications containing polyphenols are FDA-approved as pharmaceutical drugs. Polyphenon ® E, a stan