Targeting pro-senescence mitogen activated protein kinase (Mapk) enzymes with bioactive natural compounds

Depósito de investigación de la Universidad de Sevilla https://idus.us.es/ “This is an Accepted Manuscript of an article published by Elsevier in Food and Chemical Toxicology on September 2019, available at: https://doi.org/10.1016/j.fct.2019.05.052 .” 1 TARGETING PRO-AGING MITOGEN ACTIVATED PROTEIN KINASE (MAPK) ENZYMES WITH BIOACTIVE NATURAL PRODUCTS Mercedes Cano1, Angelica Guerrero-Castilla2, Antonio Ayala3 and Sandro Argüelles1* 1Department of Physiology, Faculty of Pharmacy, University of Seville, Seville, Spain; 2Química y Farmacia, Facultad de Ciencias de la Salud, Universidad Arturo Prat, Iquique, Chile; 3Department of Biochemical and Molecular Biology, Faculty of Pharmacy, University of Seville, Seville, Spain. Abstract Aging is a multifactorial universal process characterized by a gradual decrease in physiological and biochemical functions, the main risk factors contributing to age-related pathologies. Given that life expectancy is on the rise a better understanding of molecular mechanisms of the aging process is necessary in order to develop anti-aging interventions. Targeting pro-aging enzymes increases survival and therefore the lifespan. Although the up-regulation of Mitogen Activated Protein Kinases (MAPK) enzymes in aging is still controversial, increasing evidence shows that dysregulation of those enzymes are associated with biological processes that contribute to aging: including oxidative stress, inflammation, proliferation, differentiation and apoptosis. In this manuscript components of the MAPK pathway will be summarized, including extracellular signal-regulated kinase 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK) and p38, as well as several bioactive natural compounds that show positive effects on longevity, ERK, JNK and p38 enzyme inhibition. However, because a limitation of MAPK enzyme inhibitors has low specific activity and moderate toxicity, a challenge of medicinal chemistry for the pharmaceutical industry is presented in the refining of the properties of kinase enzyme inhibitors. Keywords: Aging, MAPK, ERK, JNK and p38 2 1. INTRODUCTION Aging is a multifactorial universal process that occurs at molecular, cellular and tissue levels and is characterized by a gradual decrease in physiological and biochemical functions. Recent scientific discoveries have been fundamental in understanding the molecular mechanism(s) of aging and longevity. This includes a series of mechanisms such as deregulated autophagy, mitochondrial dysfunction, shortening of telomeres, oxidative stress, systemic inflammation and metabolic dysfunction that lead the cells to a senescent state in which the cell cycle is arrested (López-Otín et al., 2013). Under normal conditions, the senescent cells release a series of proinflammatory factors, such as cytokines acting as a stimulus to be removed by the immune system, but persistent cellular senescence is a form of cell aging able to encourage the aging process by decreasing tissue renewal, repair and regeneration (Vicente et al., 2016). In fact, progressive increase of senescent cells with age has been reported to be a hallmark of aging mammalian tissues. Senescent cells secrete growth factors, proteases, and inflammatory cytokines, termed the senescence-associated secretory phenotype (SASP). An excellent review by McHugh and Gil summarized the cellular and molecular links between cellular senescence, aging and age-related diseases as well as some therapeutic approaches (McHugh and Gil, 2018). On the other hand, there is a decrease in the regenerative potential due to the reduction of stem cells in the tissues (Oh et al., 2014). Both processes contribute simultaneously to aging. One of the factors that contributes to senescence is the dysregulation of autophagy, which leads to an accumulation of cellular waste within the cell, since autophagy is a process of lysosomal degradation that cleans the cell of protein aggregates, toxic substances, aged organelles, etc. (Gewirtz, 2013). Autophagy is mainly downregulated by the mammalian target of the rapamycin (mTOR) complex that is activated under nutrient-rich conditions, but in nutrient-poor conditions, mTOR is directly inhibited by AMP-activated protein kinase (AMPK) and therefore activates the autophagy (Russell et al., 2014). During aging, there is an overexpression of the mTOR complex that leads to a deregulated autophagy, which can be inhibited by activation of AMPK (Markaki and Tavernarakis, 2013). In somatic cells, after each cell division, part of the telomere is lost, inducing telomere shortening, which is another hallmark of molecular aging (López-Otín et al., 2013). This process occurs in a limited number of cell divisions until the critical telomere size is reached, then the cells become senescent, losing the function of telomeres in the maintenance of the genome and promotion of stability in the processes of replication (Blackburn et al., 2015). In some cellular lineages, such as stem cells, telomere can be restored by the enzyme telomerase reserve transcriptase (TERT). Several studies have reported that inducing TERT activity in somatic cells reverses several characteristics of aging, such a senescent (Bär and Blasco, 2016). In 1956 Harman proposed a free radical theory of aging that suggested the role of reactive oxygen species (ROS) and oxidative stress in the process of aging are factors that aggravate the damage in cellular biomolecules. However, recent evidence shows that ROS have a double effect. They act, initially, as an activator of the homeostatic compensatory response, which increases with age and activates several antioxidant mechanisms in order to maintain survival and, from a certain level that makes injuries worse that are related to aging (Hekimi et al., 2016), suggesting that ROS effects are dependent on where they are present and their concentration. Another mechanism associated with aging is the increase of proinflammatory agents, such as tumor necrosis factor-α (TNF-α) or interleukines (IL-1 β, IL-6), which is known as “inflammaging”. Overexpression of ROS is essential for activating transcription factors, such as the activator of protein-1 (AP-1) and the nuclear factor Kappa B (NF-kB), that regulates the expression of these proinflammatory agents and contributes to the aging process (Fougère et al., 2016). In addition, mitochondrial dysfunction is associated with chronic oxidative stress in aging. The proliferator activated receptor γ coactivators α, β (PGC-1α and PGC-1β) is one of the main regulators of mitochondria. It responds to changes in nutrients status, such as the ratio of NAD+/NADH and AMP/ATP, which are regulated through SITR1 and AMPK, respectively. In aging, NAD+ levels decrease, which leads to a down regulation of SIRT1 activity inducing changes in several genes, including the decrease of PGC-1α/β, which leads to a decline in mitochondrial biogenesis (Fang et al., 2016). On the 3 other hand, AMPK, a factor that counteracts decrease in the activity of SITR1, is activated by high levels of ROS, but from certain limits the cell enters a failure in the generation of energy, creating a cellular stress and consequently leading to senescence (López-Otín et al., 2013). In general, aging is characterized by a deficiency of cellular energy fundamentally due to autophagy and mitochondrial dysfunction. Several factors are involved in this process. One of the most characteristic being the insulin/growth factor-1 (IGF-1), which in a high level is associated with aging but improves healthspam and longevity when present at low levels (Milman et al., 2014). The IGF-1 pathway regulates Forkhead box O (FOXO) transcription factors family, which have been shown to prolong the lifespan by activating genes that promote resistance to oxidative stress, DNA repair or cellular detoxification. Upon PI3K/AKT activation by IGF-1, the kinase AKT phosphorylate FOXO, promotes its cytoplasm localization and inhibits the expression of FOXO-dependent genes, which lead to a reduction of beneficial effects of the FOXO in lifespan (Martins et al., 2016). These mechanisms are all inter-connected; however, control of their processes remains unclear. Therefore, more research is necessary, specifically aimed at the molecular processes of organism aging that improves human health and lifespan. Over the years, several anti-aging interventions such as caloric restriction, telomerase and autophagy activation, as well as senolitic, epigenetic and stem cell modulators, have been studied. One of the most promising approaches to a successful anti-aging strategy includes the activation of enzymes such as AMPK, Sir1, ATG13 as potential longevity factors. Another approach to enhance survival and lifespan can be focused on the inhibitions of pro-aging enzymes such as PI3K/Akt, mTOR, P70S6k or Mitogen Activated Protein Kinases (MAPK). In this sense, the goal of this review is to analyze several bioactive natural products targeting the specific pro-aging enzymes MAPK, which play a key role in regulating important events such as oxidative stress, inflammation, proliferation, differentiation, apoptosis and stress response that all contribute to aging. 2. MITOGEN ACTIVATED PROTEIN KINASE (MAPK) AND AGING Protein phosphorylation mediated by kinase enzymes has a key role in signaling transduction pathways. Previously we summarized the relevance of the phosphorylation process mediated by kinase enzymes, as well as phosphoproteomic approaches for elucidating their molecular mechanisms in diseases (Cano et al., 2017). Mitogen activated protein kinase (MAPK) enzymes are usually regulated by phosphorylation and typically constituted by three kinases enzymes: MAPK kinase kinases (MAPKKKs), MAPK kinases (MAPKKs), and MAPKs. MAPKs pathway is an evolutionarily conserved signaling pathway associated with essential biological processes including proliferation, differentiation, apoptosis and stress response (Peti and Page, 2013; Plotnikov et al., 2011). In an excellent manuscript Plotnikov and colleagues summarized the composition, function and nuclear activities of the MAPK pathway (Plotnikov et al., 2011). In mammalian organisms the main components of the MAPK pathway are extracellular signal-regulated kinase 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38, and ERK5. Mitotic stimuli can induce the activation of ERKs pathways, while cellular/environmental stress is the most important stimuli able to activate JNK and p38 pathways (Plotnikov et al., 2011). Although the up-regulation of MAPK enzymes in aging is still controversial, increasing evidence shows that dysregulation of those enzymes are linked to the initiation and progression of the inflammation process. Chronic low grade inflammation is linked to age-associated diseases such as neurodegenerative diseases, cancer, cardiovascular diseases (CVD) and diabetes (Yeung et al., 2018). In this context, recently it has been postulated that the aging process can be a consequence of the imbalance between high activity of inflammatory pathways (inflammaging) and low activity of anti-inflammatory pathways (antiinflammaging) (Franceschi et al., 2007). In fact, increase of oxidative stress and inflammation during aging is associated with progressive loss of an appropriated immune system response (Franceschi et al., 2000). Recently, the inflammation theory of aging was summarized and contextualized by Monti and 4 colleagues. The authors showed that approaches designed to decrease and control inflammaging represent a powerful tool to modulate chronic age-related diseases and extending the healthspan of a population. Moreover, because centenaries are capable of reaching an optimal balance between inflammaging and anti-inflammaging, the authors suggest that omics data collected from those patients can contribute to the elucidation of the complex mechanism of inflammaging in elderly up to and including centenarians (Monti et al., 2017). Due to MAPK, enzymes are essential modulators of general inflammation, and of the increase inflammation that comes with age, the expectation being that MAPK enzymes are up-regulated with aging. Moreover, it could also be possible that physiological levels of MAPK enzymes contribute to the delay of irreversible senescence and while promoting longevity. However, an excessive activation of MAPK enzymes and their deficient response capacity, which might progress with aging, could lead to promotion of both irreversible cellular senescence and an accelerated aging process. This is a relevant issue because normal activation of MAPK is necessary to accurate stress response and cell survival, but their uncontrolled activation can promote inflammation and inflammatory-related pathologies, which contribute to an acceleration of the aging process. It has been reported that the general up-regulation of MAPK promotes irreversible senescence in T cells; however. different MAPK enzymes stimulate specific senescence markers. For example, pharmacological or genetic specific inhibition of p38, but not of JNK or ERK; they enhanced telomerase activity, inhibition of ERK, but not of p38 or JNK; they reduce DNA damage foci, and inhibition of JNK, but not p38 or ERK, and they promote the re-expression of the key T cell receptor signalosome component Lck (Lanna et al., 2017). The association between senescence, the aging process, age-associated diseases and regulators of initiation and progression of inflammation such as p38, JNK and ERK have been reported. 2.1 p38 MAPK pathway: The p38 MAPK pathway which is associated with the two major contributors of aging (oxidative stress and inflammation) is up-regulated in aging. In fact, Li and colleagues reported an increased regulation and activation of p38 MAPK pathway in lung and brain homogenates from a 20-month-old male rat compared to a 2-month-old young male C57BL/6J. In those mice, authors also found a concomitant increase of pro-inflammatory cytokines TNF-α, IL-1β and IL-6, and a decrease of antioxidant machinery (analyzed as superoxide dismutase (SOD)) as well as glutathione (GSH) levels (Li et al., 2011). Studies performed in the Klotho mouse models of aging showed that Klotho, an aging-suppressor gene, attenuates p38 MAPK activity and senescence pathways thereby promoting longevity (Hsieh et al., 2010). The lamins family of proteins (A/C/B) are essential components of the inner nuclear membrane and alteration of the nuclear shape is generally associated with senescence and the aging process. Using primary human fibroblasts, it was reported that upregulation of lamin B1 promoted senescence because of an increase in the percentage of SA-β-gal- positive cells, a decrease in cell proliferation and an induced alteration in chromatin structure. The authors also found that p38 MAPK activation contributes to oxidative stress-induced senescence through upregulation of lamin B1 protein (Barascu et al., 2012). Werner's syndrome (WS) is a severe premature aging pathology associated with accelerated cell senescence. Davis and colleagues reported that p38 MAPK has an important role in the reduction of accelerated cell senescence because p38 MAPK inhibitors (SB203580, BIRB 796, VX-745 and UR13756) increase growth rates, replicate capacity and reduces the stressed cellular morphology a characteristic of senesced cells (Davis et al., 2016, 2010). Several reports suggest that p38 MAPK can regulate cellular senescence and SASP (i.e., secretion of cytokines and/or chemokines) in different cell types. For example, in normal human fibroblast when p38 is activated results in the repression of human telomerase reverse the transcriptase (hTERT) gene and induction of cellular senescence (Harada et al., 2014). It has also been reported that p38 is activated in 5 response to oncogenic rasinduced premature senescence (Kwong et al., 2013; Wang et al., 2002). Similarly, high intensity ras signals induce premature senescence through activation of ERK/p38, which lead to an accumulation of cell cycle inhibitor p16INK4A (Deng et al., 2004). Hiroaki and colleagues reported that p38 MAPK has an essential role in senescent cells because p38 is activated in response to Ras-induced senescence, replicative senescence, oxidative-stress-induced senescence and inappropriate culture conditions. Authors have also shown that inhibition of p38 MAPK activation pharmacologically (SB203580) or genetically delayed the onset of senescence (Iwasa et al., 2003). It is well-known that interleukin-1beta (IL-1 β) activates p38 MAPK. Uekawa and colleagues suggested that IL-1 signal is involved in activation of p38 MAPK during senescence because in mouse embryonic fibroblasts, expression of IL-1beta increases activation of p38 and its antagonist, IL-1 receptor antagonist (IL-1Ra) negatively regulates p38 MAPK activation and senescence until IL-1β surpass IL-1Ra in expression level (Uekawa et al., 2004). Borodkina and colleagues reported that p38 is also required for H2O2-induced premature senescence of human endometrium-derived mesenchymal stem cells (hMESCs). Moreover, the authors found that inhibition of p38 MAPK with SB203580 increased proliferation of hMESCs in response to H2O2-induced premature senescence (Borodkina et al., 2014). Similarly, it has been reported that in response to oxidative stress, p38 MAPK contributes to the formation of the senescent phenotype through a mechanism involving inhibition of acetyl-CoA carboxylase 1 (ACC1), the enzyme that catalyzes the rate-limiting step in lipid synthesis (Marmisolle et al., 2017). Berry and colleagues reported a potential pharmacologic strategy to reverse cellular aging and senescence in aged beige progenitor cells. The authors found that p38 MAPK activity was enhanced in aged beige progenitors and treatment with p38 inhibitor (SB203580) reduced the expression of senescenceactivating genes and increased senescence inhibitory genes. SB203580 also reduced the number of senescence-associated betagalactosidase (SA-βgal) positive cells suggesting a rejuvenated cell (Berry et al., 2017). UVB light is a known inducer of premature senescent keratinocytes. In this context, Kim and colleagues reported that in human keratinocytes, the ability of adiponectin to reduce senescence associated markers (SA-β-gal activity, p16INK4a and phosphorylated H2AX protein expression) is linked to suppression of p38 and JNK/SAPK MAPK signaling (Kim et al., 2016). Senescent chondrocytes accumulate with age and pathogenesis of osteoarthritis. Kang and colleagues associated the effect of Cyclosporine A (CsA) and FK506 (also known as tacrolimus) as able to reduce the induction of senescence of articular chondrocytes through down-regulation of p38 MAPK because both, CSA and FK506 inhibited p38 MAPK phosphorylation (Kang et al., 2016). In addition to p38 MAPK inhibitor SB203580, which has been associated with the reduction of senescence, the two next-generation p38 MAPK inhibitors (UR-13756 and BIRB 796) with better selectivity and specificity compared to SB203580, were also able to diminish senescence-associated IL-6 secretion in human fibroblast (Alimbetov et al., 2016). Excessive exposure or accumulation of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) could induce mesenchymal stem cells (MSCs) into premature senescence. Zhou and colleagues suggested that melatonin could be a potential candidate to protect MSCs from H2O2-induced premature senescence because treatment with melatonin increased cell proliferation and proportion in the S phase, enhanced osteogenic differentiation of senescent cells, decreased SA-β-gal-positive cells and increased expression of SIRT1 while reduced expression of p16INK4α and p-p38 MAPK (L. Zhou et al., 2015). Other reports show that in response to direct DNA damage or oncogenic RAS, p38 MAPK activation promote SASP in normal human fibroblasts. The authors also found that the induction of SASP by p38 MAPK is mediated by NF-κB transcriptional activity, which was required for the expression of most SASP factors, and p38 MAPK activation during senescence can be blocked by the small molecule SB203580 (Freund et al., 2011). 6 T cell senescence contributes to immune function decline with age. Senescence in human CD8+ T cells are mediated by p38 MAPK signaling. In fact, inhibition of p38 MAPK with specific small-molecule inhibitor BIRB 796, improved telomerase activity, mitochondrial function, increased autophagy, releasing metabolic precursors and allowed the cell to increase its proliferative capacity (Henson et al., 2014). Similarly, p38 activation mediated by AMPK is associated with senescence of different cell types including T cells, fibroblasts and cardiomyocytes (Lanna et al., 2014). Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is an RNA binding protein that plays an important role in the biogenesis of mRNA, its down-regulation and its cytoplasmic accumulation contributes to a partial senescence-like morphology. Shimada and colleagues associated the regulation of hnRNP A1 with activity of the p38 MAPK pathway. In fact, inhibition of p38 MAPK activity was able to up-regulate hnRNP A1 protein expression in young rats and G0-arrested cells, as well as inhibiting cytoplasmic accumulation of hnRNP A1 in the cytoplasm of senescent cells (Shimada et al., n.d.). Up-regulation of p38 MAPK is also associated with age-related presses and pathologies. For example, (i) in modulation of endothelial senescence and cardiovascular aging, where high activity of p38 MAPK was found in senescent endothelial cells (Wu et al., 2015); (ii) sarcopenia, skeletal muscle aging can lead to sarcopenia, and poor regenerative capacities of sarcopenic muscle are associated with aged satellite cells (SCs) dysfunction. It has been found that loss of self-renewal in aged SCs was linked to up-regulation of p38 MAPK and partial inhibition of p38 MAPK with SB203580 improved aged SCs self-renewal (Bernet et al., 2014). While a growing number of reports clarify that up-regulation of p38 MAPK promotes irreversible or premature senescence and aging, and inhibition of p38 could be a potential therapeutic approach for control of aging and treatment of age-related pathologies. Other reports show that p38 MAPK activity contributes to enhanced longevity. In fact, it has been found that in Drosophila, up-regulation of p38K in muscle tissue extends life span through a mechanism involving MnSOD; however, down-regulation of p38K causes early lethality and contributes to age-related motor dysfunction and stress sensitivity (Vrailas-Mortimer et al., 2011). Similarly, down-regulation of the conserved PMK-1 p38 MAPK, which mediates the expression of secreted immune response genes, was linked to C. elegans aging because a progressive reduction in the activated PMK-1 p38 MAPK was found throughout adulthood; intact immune function stimulates longevity (Troemel et al., 2006; Youngman et al., 2011). 2.2 Jun amino-terminal kinases (JNK) MAPK pathway: Another important MAPK enzyme with a key role in oxidative stress, inflammation and that can be up-regulated in aging is JNK. In fact, a number of reports have shown an increased activity of JNK with age. Studies from the early 2000’s performed in gastric mucosa of rats reported that JNK activity increased during aging (Xiao and Majumdar, 2000). Similarly, Buzzi and colleagues found that in intestinal cells, JNK activity increased with aging (Buzzi et al., 2007). Moreover, Zhou and colleagues found an increased activity of JNKs in rat brain when compared to that from young rats (Zhou et al., 2009). D-galactose has been reported to promote senescence, oxidative stress, inflammation and apoptosis. Lian and colleagues studied the protective effect of DL0410 (a potent acetylcholinesterase inhibitor) on cognitive defects produced by aging using a D-galactose-induced aging model. The authors found that in hippocampus and cortex, D- galactose increased the expression and activation of JNK, while DL0410 was able to decrease the phosphorylation of JNK (Lian et al., 2017). Another molecule able to stimulate senescence is 5- fluorouracil (5FU). Recently it has been reported that 5FU produces senescence (assayed as increase of SA β-gal positive cell, p16INK4a and decrease of proliferation) of endothelial cells through a mechanism involving increased p38 and JNK activity as well as reduction of eNOS and SIRT-1 levels (Altieri et al., 2017). Activating transcription factor 3 (ATF3) is an important gene associated with genotoxic stress response. Kim and colleagues found that the senescence induced in macrophages by acrylamide (ACR), a 7 genotoxic carcinogen, is mediated by increased activation of p38 and JNK MAPK enzymes, which stimulate the ATF3/p53/p21 pathway leading to cellular telomerase-independent senescence (K.-H. Kim et al., 2015). Jin and colleagues studied the age-associated changes in the rat. The authors found that in the hippocampus, the age-dependent increased expression and activation of two pro-inflammatory modulators, IL-1β and JNK MAPK, were associated with decreased activation of survival pathways (Akt/p70S6K and MEK/ERK). The authors also reported that treatment with sodium ferulate, an antioxidant and anti-inflammatory compound, was able to prevent age-related changes in rats (Jin et al., 2008). The decrease of muscle mass with age, also called sarcopenia, and reduced testosterone levels are both a common hallmark of aging; thus sparking studies about the association between sarcopenia and testosterone in young and old mice. The authors found that in aged muscles, oxidative stress is increased and the expression and activation of p21 and JNK MAPK are up-regulated. The positive effect of testosterone on muscle growth in those old mice was mediated by inhibition of myostatin, p21 expression, JNK MAPK activity, activation of Akt and Notch signaling, and re-establishment of G6PDH levels (Kovacheva et al., 2010). In kidney isolated from young, middle-aged and old rats, a significant agerelated increase of oxidative stress, p38, JNK and ERK MAPK enzymes were found Using calorie restriction (CR) and intermittent fasting (IF) as anti-aging and anti-oxidative approaches, Kim and colleagues found CR to significantly decrease both the age-related activation of MAPK enzymes and the age-related oxidative stress. Moreover, Tajes and colleagues reported that IF was able to decrease JNK activity in the cortex of senescence-accelerated prone 8 (SAMP8) mice, which have a shorter lifespan (Kim et al., n.d.; Tajes et al., 2010). Another anti-aging approach is exercise. In fact, Pauli and colleagues reported that in rat skeletal muscle the increase of JNK activation with aging was reduced by acute exercise. Moreover, other age-related changes such as reduction in glucose disappearance rate and Sirt1 expression, increase in the expression of protein tyrosine phosphatase 1B, phosphorylation of IRS-1 and IkBα degradation were improved by exercise (Pauli et al., 2010). Similarly, it was reported that activity of JNK MAPK is increased in skeletal muscle from older men as compared to young men (Ghosh et al., 2015; Williamson et al., 2003), and after exercise activity of the stress-activated protein kinase JNK was reduced (Williamson et al., 2003). It is accepted that JNK MAPK is activated by stress, and stress contributes to aging and agerelated neurodegeneration. In hippocampus, which is particularly vulnerable to stress and aging, the activity of JNK was found to be strong, but not -significantly so, and it increased in old mice as compared to control. However, the JNK activity was significantly increased in the chronic mild stress aged group compared to control and aged mice, suggesting that pro-aging effects of stress could be mediated, at least in part, by JNK MAPK (Solas et al., 2013). Another structure vulnerable to stress and aging is the retina. In fact, Zhao and colleagues found a significant increase of JNK and p38 activity in aged retina tissue, showing bigger stress stimuli in aged retina rats (Zhao et al., 2014). Aging is a risk factor of increasing susceptibility to arrhythmias because in the heart the up-regulation of stress pathways gradually increases during aging. Recently, it has been reported that in the right atrial muscle of guinea pigs, the agedependent increase of JNK activity can lead to an increasing risk of atrial arrhythmias with advancing age, this risk being associated with a decrease of connexin43 (Cx43) protein expression, which is essential for preserving electrical atrial conduction (Jones and Lancaster, 2015). Similar to p38 MAPK, several studies show that JNK MAPK has an essential role in promoting cellular senescence and in contributing to an accelerated aging process. However, it has also been reported that the up-regulation of JNK MAPK activity is important in avoiding premature senescence while promoting longevity. For example, Lee and colleagues found that depletion of JNK MAPK activity 8 promotes premature senescence in breast carcinoma, lung carcinoma cancer cell line and embryonic fibroblasts. In fact, pharmacologic (SP600125) and genetic (siRNA) inhibition of JNK MAPK activity promotes premature senescence, down-regulation of Bcl2, increases ROS mitochondrial production and DNA-damage response. The authors also found that basal JNK activity is essential in preventing premature senescence and maintaining cell proliferation (Lee et al., 2010). Finally, in C. elegans and in Drosophila, JNK modulates stress resistance and promotes longevity (Biteau et al., 2011; Khoshnood et al., 2016; Oh et al., 2005). 2.3 Extracellular signal-regulated kinase (ERK) MAPK pathway: An additional MAPK enzyme with a key role in oxidative stress, inflammation and that can be up-regulated in aging is ERK. In fact, it has been reported that ERK MAPK is able to inhibit the activation of AMPK, which is essential to control the aging process and to extend both healthspan and lifespan through modulation of FoxO, mTOR and SIRT1 signaling pathways (Salminen et al., 2016). In different cell types and in response to specific stimulus, ERK MAPK activity can promote senescence in vitro and in vivo. Recently, Cagnol and Chambard showed that irregular activation of the Ras ⁄Raf ⁄ERK pathway can stimulate oncogenic transformation of immortalized cells as well as senescence of primary cells The mechanisms of ERK- induced senescence summarized by the authors include up-regulation of b-galactosidase activity and stimulation of classical senescence-associated genes, such as p16 ⁄ INK4A, p53, p21 and p14-p19 ⁄ ARF, senescence-associated heterochromatic foci and DNA damage foci (Cagnol and Chambard, 2010). Moreover, different studies have reported that tumor-suppressive action of several natural anti-oxidative drugs such as curcumin, sulforaphane and baicalin are mediated by ERK MAPK-induced senescence, because increased activation of ERK MAPK and senescence markers (e.g. increase of SA-βgal positive cells and p16INK4A protein levels, as well as decline of phosphorylation level of pRb) were found in colon cancer cells treated with these compounds (Shi et al., 2018; Zhou Wang et al., 2018). Furthermore, in rat embryo fibroblasts an increased activation of ERK MAPK was found when the senescence was induced with sodium butyrate (HDAC inhibitor) (Zhou Wang et al., 2018). Additionally, in aged B cell precursors from mice an increased activity of ERK MAPK was found (King et al., 2007). It is also known that activation of MAPK signaling is important in photo-aging. Lei and colleagues found up-regulation of MAPK enzymes including ERK, JNK and p38 phosphorylation levels during UVB-induced fibroblast senescence (Lei et al., 2017). Castello and colleagues reported an age-related increase of ERK MAPK activity in the rat heart. This up-regulation of ERK MAPK was associated with decreased STAT3 activation and SOCS3 levels. The authors also found that alternate-day fasting treatment was able to decrease ERK MAPK activity and increase STAT3 activation and SOCS3 levels (Castello et al., 2011). Similarly, other authors found an increase of ERK MAPK activity in rat aged heart lysates and heart sections (Cieslik et al., 2013), as well as in mice hearts and renal tissue but not in the kidney (Dasgupta et al., 2009). Moreover, Sieslik and colleagues reported that when mesenchymal fibroblasts, derived from aged mice hearts, were treated with AICAR, an AMPK activator, the activation of ERK MAPK decreased by 30% (Cieslik et al., 2017). Cranberry, a natural product rich in bioactive phytochemicals, can promote healthy aging. Sun and colleagues suggested that this anti-aging effect of cranberry is mediated by stress response MAPK signaling. In fact, a reduction of the accumulation of oxidative damage and down-regulation of ERK MAPK activity were found in Drosophila after supplementation with cranberry (Sun et al., 2014). Slack and colleagues reported that inhibition of ERK MAPK extends the lifespan. Authors found that both genetic (using RNAi knockdown of rl, Drosophila ortholog of Erk) and pharmacology (using trametinib, potent and highly specific inhibitor of the Mek kinase, preventing activation of Erk by Ras) inhibition of ERK activation was able to increase the lifespan in Drosophila (Slack et al., 2015). PPARβ/δ is a nuclear receptor that can inhibit skin tumorigenesis. Zhu and colleagues reported that in keratinocytes and skin 15 (GalN/LPS) 200 mg/ kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Cho et al., 2015). Human neuroblastoma cells (SH-SY5Y) treated with cytotoxicity inducer 1-methyl-4- phenylpyridinium (MTT+) 2.5 μM: ↓ p-JNK (Guifang Wang et al., 2018) ↓ Oxidative stress, ↓ Mitochondrial dysfunction, ↓ Apoptosis. Catalpol and its derivatives (2S,3R,4S,5S,6R)-2- [[(1aS,1bS,2S,5aR,6S,6a S)-6-hydroxy-1a- (hydroxymethyl)- 2,5a,6,6a-tetrahydro- 1bH- oxireno[5,6]cyclopenta[1, 3-c]pyran-2-yl]oxy]-6- (hydroxymethyl)oxane- 3,4,5-triol, a iridoid glucoside bioactive extracted from Rehmannia glutinosa Caenorhabditis elegans 25 µM: ↑28.5 % lifespan (H. W. Seo et al., 2015) Caenorhabditis elegans 20 µM: ↑17.96 % lifespan (Shukla et al., 2012) Rat pheochromocytoma cells exposed to LPS 10μM: ↓ p-p38, ↓ p-JNK (Chen et al., 2013) Epididymal adipose tissue from C57BL/6J mice with high-fat diet 100 mg/kg body weight: ↓ p-JNK (J. Zhou et al., 2015). Rotenone-induced neurotoxicity in neuron cultures from the ventral mesencephalic tissues of embryonic mice 0.01 - 0.1 mM: ↓ p-ERK, 0.02 ↓ p-JNK (Bi et al., 2009) Monocyte cell line (THP1) stimulated with phorbol- 12-myristate-13-acetate (PMA) 5 μM: ↓ p-ERK (Quan Le et al., 2015) Lung tissues from male BALB/c mice treated with LPS 2.5 – 5 – 10 mg/ kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Fu et al., 2014) ↓ Apoptosis ↓Insulin resistance, ↓ Inflammation ↓Neurotoxicity, ↓Apoptosis, ↓Necrosis ↓ Inflammation ↓ Inflammation 16 Chlorogenic acid (1S,3R,4R,5R)-3-[(E)-3- (3,4-dihydroxyphenyl) prop-2-enoyl]oxy-1,4,5- trihydroxycyclohexane-1- carboxylic acid, is the major polyphenolic compound in coffee, isolated from the leaves and fruits of dicotyledonous plants. Caenorhabditis elegans 50 µM: ↑27.1 % lifespan (S.-Q. Zheng et al., 2016) Liver tissue from male ICR mice with polymicrobial sepsis induced 20 mg/kg body weight: ↓ p-p38, ↓ p-JNK (Lee et al., 2013) Sprague-Dawley rats with hepatic sinusoidal obstruction syndrome 20 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Z. Zheng et al., 2016). Acetaminophen-induced liver injury in ICR mice 40 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Ji et al., 2013). CCl4 and platelet derived growth factor (PDGF)- induced liver fibrosis in rat hepatic stellate cells (HSC-T6) 50 µg/mL: ↓ p-ERK, ↓ p-p38 (Shi et al., 2016). Phorbol-12-myristate-13- acetate PMA- treated human lung adenocarcinoma (A549) cells 150 μM: ↓ p-ERK 200 μM: ↓ p-JNK (Tsai et al., 2013). RANKL-mediated osteoclast differentiation in bone marrow macrophages 10, 25 and 50 μg/ml: ↓ p-p38 25 and 50 μg/ml: ↓ p-ERK 25 and 50 μg/ml: ↓ p-JNK ↓ Inflammation, ↓ Apoptosis ↓ Inflammation, ↑Antioxidants ↓ Inflammation, ↓ Apoptosis ↑ Antioxidants, ↓ Oxidative stress ↓ Inflammation, ↓ Invasion ↓Inflammationinduced bone loss, ↓Osteoclasts differentiation 17 (Kwak et al., 2013). JB6 Murine epidermal cell exposed to UVB 15 μM: ↓ p-p38, ↓ p-JNK 140 μM: ↓ p-ERK (Feng et al., 2005) Mouse Bone Marrow Macrophage (BMMs) treated with nuclear factor-kappa B (NF-κB) ligand. 10 – 25 – 50 µg/mL: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Kwak et al., 2013) Concanavalin A-induced hepatitis in Balb/c mice 30 mg/kg body weight: ↓ p-p38 (Yuan et al., 2017). H2O2-induced apoptosis in H9c2 cardiomyoblasts cells 12.5 - 25 μM: ↓ p-ERK, ↓ p-JNK (Yu et al., 2016). LPS-activated murine macrophage-like cells (RAW264.7) 37.5 µg/mL: ↓ p-JNK (Shan et al., 2009). Rat pheochromocytoma (PC12) cell line treated with H2O2 1- 5 μM: ↓ p-p38, ↓ p-JNK (Cho et al., 2009). VEGF- activated primary human retinal endothelial cells 2.5 μM: ↓ p-ERK, ↓ p-p38 (Mei et al., 2018). ↓ Inflammation, ↓ Carcinogenesis, ↑Antioxidants ↓ Inflammatory bone destruction ↑ Liver function, ↓ Liver inflammatory responses. ↓ Oxidative stress, ↓ Apoptosis. ↓ Inflammation. ↓ Oxidative stress, ↓ Apoptosis. ↓ Retinal neoangiogenesis, ↓ Inflammation. 18 Ellagic Acid 2,3,7,8- Tetrahydroxychromeno [5,4,3-cde]chromene- 5,10-dione, is a fused four ring compound occurring free or combined in galls. Isolated from the kino of Eucalyptus maculata and Eucalyptus Hemipholia. Caenorhabditis elegans 50 µM: ↑10 % lifespan (Saul et al., 2011). Buccal pouch carcinomas induced by 7,12 – dimethylbenz [a] anthracene in Male Syrian hamsters 0.4% in the diet: ↓ p-ERK, ↓ p-JNK (Kowshik et al., 2014). LPS-induced RAW 264.7 cells 0.3 µM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK 1.0 µM: ↓ p-ERK, ↓ p-p38 (J. Lee et al., 2014) Dextran sulfate sodiuminduced acute and chronic colitis in Balb/C mice 0.5% in the diet: ↓ p-p38 (Marín et al., 2013). Concanavalin A-induced hepatitis in BALB/c mice 50 – 100 - 200 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (J. H. Lee et al., 2014). Collagen-activated Human platelet 50 - 80 µM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Chang et al., 2013). ↓ Tumor angiogenesis, ↓ Neovascularization ↓ Inflammation ↓ Inflammation ↓ Liver necrosis, ↓ Inflammation ↓ Platelet aggregation, ↓ ROS Epigallocatechin-3- gallate (EGCG) [(2R,3R)-5,7-dihydroxy- 2-(3,4,5- trihydroxyphenyl)-3,4- dihydro-2H-chromen-3- yl] 3,4,5- trihydroxybenzoate, is a phenolic antioxidant found in a number of plants such as green and black tea. Caenorhabditis elegans 100 µM: ↑20 % lifespan (Bartholome et al., 2010). Caenorhabditis elegans 0.1µg/mL: ↑13.1% lifespan under heat stress and ↑172.9% lifespan under oxidative stress (Zhang et al., 2009). Wistar rats 25 mg /kg: ↑14.1 % lifespan Thrombin-induced neuron degeneration in cortical neuronal cell culture 25 µM: ↓ p-JNK (Q. He et al., 2015). TGF-b-induced myofibroblast differentiation in prostate fibroblast cell line 40 µM: ↓ p-ERK (Gray et al., 2014). Myocardial ischemia ↑ Neuroprotecction, ↓ Apoptosis ↓ Tumor myofibroblast differentiation, ↓ Tumor cell invasion ↓ Myocardial I/R 19 (Niu et al., 2013) induced in Male Sprague– Dawley rats 10 mg/kg body weight: ↓ p-p38, ↓ p-JNK (S. J. Kim et al., 2014). Toxicity induced by nickel nanoparticles (Ni NPs) in mouse epidermal cell line 10 µM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (In the 2.5 and 5 μg/cm2 Ni NPs treatment groups). (Gu et al., 2016) LPS-stimulated human acute monocytic leukemia cell line 50 μg/ml: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (WANG et al., 2014) Human retinal endothelial cells treated with high glucose concentrations 20 - 40 µM: ↓ p-ERK, ↓ p-p38 (Zhang et al., 2016) LPS-induced innate signaling in bone marrowderived macrophages isolated from ICR mice 100 µM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Joo et al., 2012). Homocysteine -induced proliferation of vascular smooth muscle cells 20 µM: ↓ p-ERK, ↓ p-p38 (Zhan et al., 2018). High fat and high-fructose diet-induced body weight gain in C57BL/6J mice 2 g/L: ↓ p-p38 (Mi et al., 2017). injuries ↓ Oxidative stress, ↓ Apoptosis. ↓ Inflammation ↓ Inflammation, ↓ Apoptosis ↓ Inflammation ↓ Proliferation ↓ Memory impairment, ↓ Neuronal loss, ↓ Inflammation ↓ Oxidative stress, ↓ Mitochondrial dysfunction 20 Pancreatic alpha TC1-6 (αTC1-6) cells exposed to H2O2 100 µM: ↓ p-p38, ↓ p-JNK (Cao et al., 2018). ↓ Oxidative stress, ↓ Apoptosis ↓ Dysfunction of glucagon secretion Fisetin 2-(3,4-dihydroxyphenyl)- 3,7-dihydroxychromen-4- one, is a plant polyphenol from the flavonoid group. It can be found in many plants, fruits and vegetables, such as strawberries, apples, persimmons, onions and cucumbers. Drosophila melanogaster 100 µM: ↑23 % lifespan (Wood et al., 2004). Saccharomyces cerevisiae 10 μM: ↑55% lifespan (Howitz et al., 2003). LPS- treated Human gingival fibroblasts 1 – 5- 10 - 15 µM: ↓ p-p38, ↓ p-JNK (Gutiérrez-Venegas et al., 2014). 12-O-tetradecanoyl- phorbol-13-acetate TPA- induced cell invasion in MCF-7 human breast cancer cells 30 µM: ↓ p-ERK, ↓ p-p38 (Noh et al., 2015). Melanoma cells (A375 and RPMI-7951) 5 – 10 - 20 µM: ↓ p-ERK (Pal et al., 2014) Human cervical adenocarcinoma (SiHa and CaSki cells) 20 – 40 µM: ↓ p-p38 (Chou et al., 2013). Primary astrocyte cultures from 1-day-old Sprague- Dawley rat brains 12.5 – 25 – 50 µM: ↓ p-ERK (N. Wang et al., 2017). Dextran sulphate sodiuminduced colitis in male Balb/C mice 5 and 10 mg/kg body weight: ↓ p-p38 (Sahu et al., 2016). ↓ Inflammation ↓ Cell invasion, ↓ Inflammation ↓ Cell invasion, ↑ epithelialmesenchymal transition, ↓ Cell invasion, ↓ Migration ↓ Cell proliferation, ↓ Migration ↓ Inflammation, ↑ Antioxidants, ↓ Oxidative stress Fucoxanthin [(1S,3R)-3-hydroxy-4- Drosophila melanogaster 0.3 – 1.0 µM: LPS-stimulated murine macrophage RAW 264.7 ↓ Inflammation, ↓ Oxidative stress 21 [(3E,5E,7E,9E,11E,13E,1 5E)-18-[(1R,3S,6S)-3- hydroxy-1,5,5-trimethyl- 7-xabicyclo[4.1.0]heptan- 6-yl]-3,7,12,16- tetramethyl-17- oxooctadeca- 1,3,5,7,9,11,13,15- octaenylidene]-3,5,5- trimethylcyclohexyl] acetate, is a marine carotenoid found in numerous classes of microalgae and brown macroalgae ↑33 % lifespan (Male) ↑33-49 % lifespan (Female) (Lashmanova et al., 2015) Drosophila melanogaster 1 µM: ↑14.9 % lifespan (Male); ↑6.2% lifespan (Female) (Moskalev et al., 2018). cells (Kim et al., 2010) 12.5 - 25 and 50 μM: ↓ p-ERK, ↓ p-p38, ↓ p- JNK Amyloid-β42-stimulated murine BV2 microglia cells 5 - 10 and 50 μM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Pangestuti et al., 2013). Human colorectal cancer cells under hypoxic conditions 50 μM: ↓ p-ERK (Terasaki et al., 2018) Human hepatoma cells treated with cisplatin 10 μM: ↓ p-ERK, ↓ p-p38 (Liu et al., 2013) Human Neuroblastoma cells (SH-SY5Y) exposed to H2O2 1 - 3 μM: ↓ p-ERK (Yu et al., 2017). β-amyloid –stimulated SH-SY5Y cells 1 - 3 μM: ↓ p-ERK (Lin et al., 2017). ↓ Inflammation, ↓ Oxidative stress, ↓ Microglial activation ↓ Colonospheres formation, ↑ Apoptosis ↓ Epithelialmesenchymal transition ↓ Proliferation, ↑ Apoptosis ↓ Oxidative stress, ↓ Apoptosis ↓ Oxidative stress, ↓ Apoptosis Gallic acid and its derivatives 3,4,5-trihydroxybenzoic acid, is an organic acid found in gallnuts, sumac, witch hazel, tea leaves, oak bark, and other plants. Caenorhabditis elegans 300 µM: ↑10 % lifespan (Saul et al., 2011). Caenorhabditis elegans *Pentagalloyl glucose 160 μM: ↑18% lifespan (Y. Chen et al., 2014) High fat diet/ streptozotocin-induced Diabetes in wistar albino rats 25 and 50 mg/kg body weight: ↓ p-p38 (Ahad et al., 2015). Lung cancer cell (A549) 100 µM: ↓ p-p38, ↓ p-JNK (PARK and KIM, 2013) Human glioblastoma U87 and U251n ↑ Nephroprotection, ↓ Blood glucose, ↓ Inflammation ↓ Proliferation, ↑ Apoptosis, ↓ Antioxidant (GSH) ↓ Cell viability, ↓ Proliferation, 22 30 – 40 µM: ↓ p-ERK (Lu et al., 2010). Human stimulated platelets 100 - 500 µM: ↓ p-p38 (Chang et al., 2012). Stress-induced in rat pheochromacytoma (PC12) cell lines 1 µM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Huang et al., 2012). The human nasopharyngeal carcinoma cells (NPC-BM1) 50 µM: ↓ p-p38 (Pang et al., 2017). LPS- stimulated microglial cells (BV-2) 1 µM: ↓ p-JNK (W.-H. Lin et al., 2015). Human cervical cancer HeLa and HTB-35 cells 10 – 15 - 20 µg/mL: ↓ p-ERK (Zhao and Hu, 2013). ↓ Invasion ↓ Platelet activation, ↓ Platelet-leukocyte aggregation ↓ Inflammation, ↓ Oxidative stress ↓ Cell invasion, ↓ Migration ↓ Inflammation, ↓ Oxidative stress ↓ Proliferation, ↓ Invasion, ↓ Angiogenesis Genistein 5,7-dihydroxy-3-(4- hydroxyphenyl)chromen- 4-one, is an isoflavonoid found in a number of plants, but soybeans and soy products like tofu and textured vegetable protein are the primary food source. Caenorhabditis elegans 100 μM: ↑28% lifespan 50 μM: ↑10% lifespan (Lee et al., 2015). d-Galactosamine-induced inflammation and hepatotoxicity in male Wistar rats 5 mg/kg body weight: ↓ p-ERK, ↓ p-p38 (Ganai et al., 2015). H2O2-induced cell death in primary rat cortical neurons 1 µM: ↓ p-ERK, ↓ p-JNK (Qian et al., 2015) VEGF- activated Human Umbilical Vein Endothelial Cells 100 µM: ↓ p-p38, ↓ p-JNK ↑ Hepatoprotection, ↓ Histological damage, ↓ Inflammation, ↓ Oxidative stress ↓ Inflammation, ↓ Oxidative stress, ↓ Apoptosis ↓ Angiogenesis, ↑ Apoptosis 23 (Yu et al., 2012). Cardiac hypertrophy in C57/BL6 mice and phenylephrine-induced hypertrophy in neonatal rat cardiomyocytes 40 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (in vivo assay- Mice) 20 µM: ↓ p-JNK (in vitro assay –NRCM) (Meng et al., 2017). Lead (Pb)-induced toxicity in PC12 cell line 10 µM: ↓ p-ERK, ↓ p-p38 (Su et al., 2016). Non-alcoholic steatohepatitis-induced by high fat diet in male Sprague–Dawley rats 4 - 8 mg/kg body weight: ↓ p-JNK (Ji et al., 2011). PMA/A23187-induced mast cell activation (HMC-1 cells) 50 µM: ↓ p-ERK (KIM et al., 2014). ↓ Endomyocardial fibrosis, ↓ Hypertrophic stimulation ↓ Apoptosis, ↑Antioxidant ↓ Liver damage, ↓ Inflammation, ↓ Oxidative stress, ↓ Inflammation Quercetin 2-(3,4-dihydroxyphenyl)- 3,5,7-trihydroxychromen- 4-one, is a polyphenolic flavonoid widely distributed in many plants and fruits including red grapes, citrus fruit, tomato, broccoli and other leafy green vegetables, and a number of berries, including raspberries and cranberries. Podospora anserine 300 μM: ↑10.2% lifespan (Warnsmann et al., 2018). Caenorhabditis elegans 200 μM (quercetin-3-O- glucoside): ↑39% lifespan (Dueñas et al., 2013). Caenorhabditis elegans 200 μM: ↑11% lifespan (Surco-Laos et al., 2011). Caenorhabditis elegans 100 μM: ↑11% lifespan 200 μM: ↑18% lifespan (Pietsch et al., 2009). Chronic Prostatitis /Chronic Pelvic Pain Syndrome in Sprague Dawley rats 50 – 100 – 200 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Meng et al., 2018). Adipogenesis induced by 3-isobutyl-1- methylxanthine and dexamethasone in preadipocyte cells 12.5 - 25 μM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (M.-J. Seo et al., 2015). ↓ Inflammation, ↓ Oxidative stress ↓ Lipid accumulation, ↓ Obesity-induced inflammation 24 Drosophila melanogaster 1μM: ↑13% lifespan (Proshkina et al., 2016) Nontypeable Haemophilus influenza-stimulated Human middle ear epithelial cells 80 μM: ↓ p-p38 (Ma et al., 2018). Chymotrypsin-like activity of proteasome in HepG2 cells 25 – 50 - 100 μM: ↓ p-ERK (Ding et al., 2018). VEGF-stimulated Mouse cone photoreceptorderived 661W cells 0.1 µM: ↓ p-ERK, 0.2 ↓ p-p38, ↓ p-JNK (Lee et al., 2017). Liver injury induced by intraperitoneal injection of LPS (50 μg/kg)/D-GalN (300 mg/kg) in BALB/c Kunming species mice 50 – 100 mg/kg body weight: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Peng et al., 2017). LPS-stimulated Myoblast rat H9c2 cells 10 μM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Gutiérrez-Venegas et al., 2017). LPS-stimulated Murine macrophage RAW 264.7 cell line 50 μM: ↓ p-ERK, ↓ p-p38, ↓ p-JNK (Cho et al., 2016). Renal toxicity by Cisplatin-induced in Male Fischer F344 rats 50 mg/kg body weight: ↓ Inflammation ↓ Proteasome activity, ↓ Proliferation ↓ Inflammation, ↓ Oxidative stress ↓ Apoptosis, ↓ Inflammation, ↓ Oxidative stress ↓ Inflammation ↓ Inflammation ↓ Cisplatin nephrotoxicity, ↓ Inflammation, ↓ Apoptosis, 31 Cho, Y.-H., Kim, N.-H., Khan, I., Yu, J.M., Jung, H.G., Kim, H.H., Jang, J.Y., Kim, H.J., Kim, D., Kwak, J.-H., Kang, S.C., An, B.J., 2016. Anti-inflammatory Potential of Quercetin-3-O-β-D-(“2”-galloyl)- glucopyranoside and Quercetin Isolated from Diospyros kaki calyx via Suppression of MAP Signaling Molecules in LPS-induced RAW 264.7 Macrophages. J. Food Sci. 81, C2447–C2456. doi:10.1111/1750-3841.13497 Chou, R.-H., Hsieh, S.-C., Yu, Y.-L., Huang, M.-H., Huang, Y.-C., Hsieh, Y.-H., 2013. Fisetin inhibits migration and invasion of human cervical cancer cells by down-regulating urokinase plasminogen activator expression through suppressing the p38 MAPK-dependent NF-κB signaling pathway. PLoS One 8, e71983. doi:10.1371/journal.pone.0071983 Cieslik, K.A., Trial, J., Carlson, S., Taffet, G.E., Entman, M.L., 2013. Aberrant differentiation of fibroblast progenitors contributes to fibrosis in the aged murine heart: role of elevated circulating insulin levels. FASEB J. 27, 1761–71. doi:10.1096/fj.12-220145 Cieslik, K.A., Trial, J., Entman, M.L., 2017. Aicar treatment reduces interstitial fibrosis in aging mice. J. Mol. Cell. Cardiol. 111, 81–85. doi:10.1016/j.yjmcc.2017.08.003 Cui, Y.-Q., Wang, Q., Zhang, D.-M., Wang, J.-Y., Xiao, B., Zheng, Y., Wang, X.-M., 2016. Triptolide Rescues Spatial Memory Deficits and Amyloid-β Aggregation Accompanied by Inhibition of Inflammatory Responses and MAPKs Activity in APP/PS1 Transgenic Mice. Curr. Alzheimer Res. 13, 288–96. Danilov, A., Shaposhnikov, M., Shevchenko, O., Zemskaya, N., Zhavoronkov, A., Moskalev, A., 2015. Influence of non-steroidal anti-inflammatory drugs on Drosophila melanogaster longevity. Oncotarget 6, 19428–44. doi:10.18632/oncotarget.5118 Dasgupta, J., Kar, S., Van Remmen, H., Melendez, J.A., 2009. Age-dependent increases in interstitial collagenase and MAP Kinase levels are exacerbated by superoxide dismutase deficiencies. Exp. Gerontol. 44, 503–10. doi:10.1016/j.exger.2009.04.005 Davis, T., Bachler, M.A., Wyllie, F.S., Bagley, M.C., Kipling, D., 2010. Evaluating the role of p38 MAP kinase in growth of Werner syndrome fibroblasts. Ann. N. Y. Acad. Sci. 1197, 45–48. doi:10.1111/j.1749-6632.2010.05195.x Davis, T., Brook, A., Rokicki, M., Bagley, M., Kipling, D., 2016. Evaluating the Role of p38 MAPK in the Accelerated Cell Senescence of Werner Syndrome Fibroblasts. Pharmaceuticals 9, 23. doi:10.3390/ph9020023 Deng, Q., Liao, R., Wu, B.-L., Sun, P., 2004. High intensity ras signaling induces premature senescence by activating p38 pathway in primary human fibroblasts. J. Biol. Chem. 279, 1050–9. doi:10.1074/jbc.M308644200 Ding, Y., Chen, X., Wang, B., Yu, B., Ge, J., Shi, X., 2018. Quercetin suppresses the chymotrypsin-like activity of proteasome via inhibition of MEK1/ERK1/2 signaling pathway in hepatocellular carcinoma HepG2 cells. Can. J. Physiol. Pharmacol. 96, 521–526. doi:10.1139/cjpp-2017-0655 Donato, A.J., Morgan, R.G., Walker, A.E., Lesniewski, L.A., 2015. Cellular and molecular biology of aging endothelial cells. J. Mol. Cell. Cardiol. 89, 122–135. doi:10.1016/j.yjmcc.2015.01.021 Dueñas, M., Surco-Laos, F., González-Manzano, S., González-Paramás, A.M., Gómez-Orte, E., Cabello, J., Santos-Buelga, C., 2013. Deglycosylation is a key step in biotransformation and lifespan effects of quercetin-3-O-glucoside in Caenorhabditis elegans. Pharmacol. Res. 76, 41–8. doi:10.1016/j.phrs.2013.07.001 Fang, E.F., Scheibye-Knudsen, M., Chua, K.F., Mattson, M.P., Croteau, D.L., Bohr, V.A., 2016. Nuclear 32 DNA damage signalling to mitochondria in ageing. Nat. Rev. Mol. Cell Biol. 17, 308–321. doi:10.1038/nrm.2016.14 Feng, R., Lu, Y., Bowman, L.L., Qian, Y., Castranova, V., Ding, M., 2005. Inhibition of activator protein-1, NF-kappaB, and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid. J. Biol. Chem. 280, 27888–95. doi:10.1248/bpb.b13-00430 Fong, Y., Shen, K.H., Chiang, T.A., Shih, Y.W., 2010. Acacetin inhibits TPA-induced MMP-2 and u-PA expressions of human lung cancer cells through inactivating JNK signaling pathway and reducing binding activities of NF-κB and AP-1. J. Food Sci. 75, 30–38. doi:10.1111/j.1750- 3841.2009.01438.x Fougère, B., Boulanger, E., Nourhashémi, F., Guyonnet, S., Cesari, M., 2016. Chronic Inflammation: Accelerator of Biological Aging. Journals Gerontol. Ser. A Biol. Sci. Med. Sci. 72, glw240. doi:10.1093/gerona/glw240 Franceschi, C., Bonafè, M., Valensin, S., Olivieri, F., De Luca, M., Ottaviani, E., De Benedictis, G., 2000. Inflamm-aging. An evolutionary perspective on immunosenescence. Ann. N. Y. Acad. Sci. 908, 244–54. Franceschi, C., Capri, M., Monti, D., Giunta, S., Olivieri, F., Sevini, F., Panourgia, M.P., Invidia, L., Celani, L., Scurti, M., Cevenini, E., Castellani, G.C., Salvioli, S., 2007. Inflammaging and antiinflammaging: a systemic perspective on aging and longevity emerged from studies in humans. Mech. Ageing Dev. 128, 92–105. doi:10.1016/j.mad.2006.11.016 Freund, A., Patil, C.K., Campisi, J., 2011. p38MAPK is a novel DNA damage response-independent regulator of the senescence-associated secretory phenotype. EMBO J. 30, 1536–1548. doi:10.1038/emboj.2011.69 Fu, K., Piao, T., Wang, M., Zhang, J., Jiang, J., Wang, X., Liu, H., 2014. Protective effect of catalpol on lipopolysaccharide-induced acute lung injury in mice. doi:10.1016/j.intimp.2014.07.011 Ganai, A.A., Khan, A.A., Malik, Z.A., Farooqi, H., 2015. Genistein modulates the expression of NF-κB and MAPK (p-38 and ERK1/2), thereby attenuating d-Galactosamine induced fulminant hepatic failure in Wistar rats. Toxicol. Appl. Pharmacol. 283, 139–146. doi:10.1016/j.taap.2015.01.012 Gao, Y., Kang, L., Li, C., Wang, X., Sun, C., Li, Q., Liu, R., Wang, J., 2016. Resveratrol Ameliorates Diabetes-Induced Cardiac Dysfunction Through AT1R-ERK/p38 MAPK Signaling Pathway. Cardiovasc. Toxicol. 16, 130–137. doi:10.1007/s12012-015-9321-3 Ge, G.F., Shi, W.W., Yu, C.H., Jin, X.Y., Zhang, H.H., Zhang, W.Y., Wang, L.C., Yu, B., 2017. Baicalein attenuates vinorelbine-induced vascular endothelial cell injury and chemotherapeutic phlebitis in rabbits. Toxicol. Appl. Pharmacol. 318, 23–32. doi:10.1016/j.taap.2017.01.013 Gewirtz, D.A., 2013. Autophagy and senescence. Autophagy 9, 808–812. doi:10.4161/auto.23922 Ghosh, S., Lertwattanarak, R., Garduño, J. de J., Galeana, J.J., Li, J., Zamarripa, F., Lancaster, J.L., Mohan, S., Hussey, S., Musi, N., 2015. Elevated muscle TLR4 expression and metabolic endotoxemia in human aging. J. Gerontol. A. Biol. Sci. Med. Sci. 70, 232–46. doi:10.1093/gerona/glu067 Gray, A.L., Stephens, C.A., Bigelow, R.L.H., Coleman, D.T., Cardelli, J.A., 2014. The polyphenols (-)- epigallocatechin-3-gallate and luteolin synergistically inhibit TGF-β-induced myofibroblast phenotypes through RhoA and ERK inhibition. PLoS One 9, e109208. doi:10.1371/journal.pone.0109208 33 Gruber, J., Tang, S.Y., Halliwell, B., 2007. Evidence for a trade-off between survival and fitness caused by resveratrol treatment of Caenorhabditis elegans. Ann. N. Y. Acad. Sci. 1100, 530–42. doi:10.1196/annals.1395.059 Gu, Y., Wang, Y., Zhou, Q., Bowman, L., Mao, G., Zou, B., Xu, J., Liu, Y., Liu, K., Zhao, J., Ding, M., 2016. Inhibition of Nickel Nanoparticles-Induced Toxicity by Epigallocatechin-3-Gallate in JB6 Cells May Be through Down-Regulation of the MAPK Signaling Pathways. PLoS One 11, e0150954. doi:10.1371/journal.pone.0150954 Guo, R., Su, Y., Liu, B., Li, S., Zhou, S., Xu, Y., 2014. Resveratrol suppresses oxidised low-density lipoprotein-induced macrophage apoptosis through inhibition of intracellular reactive oxygen species generation, LOX-1, and the p38 MAPK pathway. Cell. Physiol. Biochem. 34, 603–16. doi:10.1159/000363026 Gutiérrez-Venegas, G., Contreras-Sánchez, A., Ventura-Arroyo, J.A., 2014. Anti-inflammatory activity of fisetin in human gingival fibroblasts treated with lipopolysaccharide. J. Asian Nat. Prod. Res. 16, 1009–1017. doi:10.1080/10286020.2014.932351 Gutiérrez-Venegas, G., Torras-Ceballos, A., Gómez-Mora, J.A., Fernández-Rojas, B., 2017. Luteolin, quercetin, genistein and quercetagetin inhibit the effects of lipopolysaccharide obtained from Porphyromonas gingivalis in H9c2 cardiomyoblasts. Cell. Mol. Biol. Lett. 22, 19. doi:10.1186/s11658-017-0047-z Ha, S.K., Moon, E., Lee, P., Ryu, J.H., Oh, M.S., Kim, S.Y., 2012. Acacetin attenuates neuroinflammation via regulation the response to LPS stimuli in Vitro and in Vivo. Neurochem. Res. 37, 1560–1567. doi:10.1007/s11064-012-0751-z Han, S.-Y., Choi, Y.-J., Kang, M.-K., Park, J.H.Y., Kang, Y.-H., 2015. Resveratrol Suppresses Cytokine Production Linked to FcεRI-MAPK Activation in IgE-Antigen Complex-Exposed Basophilic Mast Cells and Mice. Am. J. Chin. Med. 43, 1605–1623. doi:10.1142/S0192415X15500913 Harada, G., Neng, Q., Fujiki, T., Katakura, Y., 2014. Molecular mechanisms for the p38-induced cellular senescence in normal human fibroblast. J. Biochem. 156, 283–290. doi:10.1093/jb/mvu040 Havermann, S., Humpf, H.U., Wätjen, W., 2016. Baicalein modulates stress-resistance and life span in C. elegans via SKN-1 but not DAF-16. Fitoterapia 113, 123–127. doi:10.1016/j.fitote.2016.06.018 Havermann, S., Rohrig, R., Chovolou, Y., Humpf, H., 2013. of Lifespan. doi:10.1021/jf304553g. He, L., Liang, Z., Zhao, F., Peng, L., Chen, Z., 2015. Modulation of IL-37 expression by triptolide and triptonide in THP-1 cells. Cell. Mol. Immunol. 12, 515–8. doi:10.1038/cmi.2014.92 He, Q., Bao, L., Zimering, J., Zan, K., Zhang, Z., Shi, H., Zu, J., Yang, X., Hua, F., Ye, X., Cui, G., 2015. The protective role of (-)-epigallocatechin-3-gallate in thrombin-induced neuronal cell apoptosis and JNK-MAPK activation. Neuroreport 26, 416–23. doi:10.1097/WNR.0000000000000363 He, W., Shi, F., Zhou, Z.-W., Li, B., Zhang, K., Zhang, X., Ouyang, C., Zhou, S.-F., Zhu, X., 2015. A bioinformatic and mechanistic study elicits the antifibrotic effect of ursolic acid through the attenuation of oxidative stress with the involvement of ERK, PI3K/Akt, and p38 MAPK signaling pathways in human hepatic stellate cells and rat liver. Drug Des. Devel. Ther. 9, 3989–4104. doi:10.2147/DDDT.S85426 He, X., Wei, Z., Zhou, E., Chen, L., Kou, J., Wang, J., Yang, Z., 2015. Baicalein attenuates inflammatory responses by suppressing TLR4 mediated NF-κB and MAPK signaling pathways in LPS-induced mastitis in mice. Int. Immunopharmacol. 28, 470–476. doi:10.1016/j.intimp.2015.07.012 34 Hekimi, S., Wang, Y., Noë, A., 2016. Mitochondrial ROS and the Effectors of the Intrinsic Apoptotic Pathway in Aging Cells: The Discerning Killers! Front. Genet. 7, 161. doi:10.3389/fgene.2016.00161 Henson, S.M., Lanna, A., Riddell, N.E., Franzese, O., Macaulay, R., Griffiths, S.J., Puleston, D.J., Watson, A.S., Simon, A.K., Tooze, S.A., Akbar, A.N., 2014. p38 signaling inhibits mTORC1- independent autophagy in senescent human CD8+ T cells. J. Clin. Invest. 124, 4004–16. doi:10.1172/JCI75051 Heo, J.-R., Lee, G.-A., Kim, G.-S., Hwang, K.-A., Choi, K.-C., 2018. Phytochemical-induced reactive oxygen species and endoplasmic reticulum stress-mediated apoptosis and differentiation in malignant melanoma cells. Phytomedicine 39, 100–110. doi:10.1016/j.phymed.2017.12.006 Howitz, K.T., Bitterman, K.J., Cohen, H.Y., Lamming, D.W., Lavu, S., Wood, J.G., Zipkin, R.E., Chung, P., Kisielewski, A., Zhang, L.-L., Scherer, B., Sinclair, D.A., 2003. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature 425, 191–196. doi:10.1038/nature01960 Hsieh, C.-C., Kuro-o, M., Rosenblatt, K.P., Brobey, R., Papaconstantinou, J., 2010. The ASK1- Signalosome regulates p38 MAPK activity in response to levels of endogenous oxidative stress in the Klotho mouse models of aging. Aging (Albany. NY). 2, 597–611. doi:10.18632/aging.100194 Hu, S., Chen, Y., Wang, Z.-F., Mao-Ying, Q.-L., Mi, W.-L., Jiang, J.-W., Wu, G.-C., Wang, Y.-Q., 2015. The Analgesic and Antineuroinflammatory Effect of Baicalein in Cancer-Induced Bone Pain. Evid. Based. Complement. Alternat. Med. 2015, 973524. doi:10.1155/2015/973524 Huang, H.-L., Lin, C.-C., Jeng, K.-C.G., Yao, P.-W., Chuang, L.-T., Kuo, S.-L., Hou, C.-W., 2012. Fresh green tea and gallic acid ameliorate oxidative stress in kainic acid-induced status epilepticus. J. Agric. Food Chem. 60, 2328–36. doi:10.1021/jf203709q Ikeyama, S., Kokkonen, G., Shack, S., Wang, X.-T., Holbrook, N.J., 2002. Loss in oxidative stress tolerance with aging linked to reduced extracellular signal-regulated kinase and Akt kinase activities. FASEB J. 16, 114–6. doi:10.1096/fj.01-0409fje Iwasa, H., Han, J., Ishikawa, F., 2003. Mitogen-activated protein kinase p38 defines the common senescence-signalling pathway. Genes Cells 8, 131–44. Ji, G., Yang, Q., Hao, J., Guo, L., Chen, X., Hu, J., Leng, L., Jiang, Z., 2011. Anti-inflammatory effect of genistein on non-alcoholic steatohepatitis rats induced by high fat diet and its potential mechanisms. Int. Immunopharmacol. 11, 762–768. doi:10.1016/j.intimp.2011.01.036 Ji, L., Jiang, P., Lu, B., Sheng, Y., Wang, X., Wang, Z., 2013. Chlorogenic acid, a dietary polyphenol, protects acetaminophen-induced liver injury and its mechanism. J. Nutr. Biochem. 24, 1911–1919. doi:10.1016/j.jnutbio.2013.05.007 Jin, Y., Yan, E., Li, X., Fan, Y., Zhao, Y., Liu, Z., Liu, W., 2008. Neuroprotective effect of sodium ferulate and signal transduction mechanisms in the aged rat hippocampus. Acta Pharmacol. Sin. 29, 1399–408. doi:10.1111/j.1745-7254.2008.00848.x Jones, S.A., Lancaster, M.K., 2015. Progressive age-associated activation of JNK associates with conduction disruption in the aged atrium. Mech. Ageing Dev. 146–148, 72–80. doi:10.1016/j.mad.2015.05.001 Joo, S.-Y., Song, Y.-A., Park, Y.-L., Myung, E., Chung, C.-Y., Park, K.-J., Cho, S.-B., Lee, W.-S., Kim, H.-S., Rew, J.-S., Kim, N.-S., Joo, Y.-E., 2012. Epigallocatechin-3-gallate Inhibits LPS-Induced NF-κB and MAPK Signaling Pathways in Bone Marrow-Derived Macrophages. Gut Liver 6, 188– 96. doi:10.5009/gnl.2012.6.2.188 35 Jurk, D., Wilson, C., Passos, J.F., Oakley, F., Correia-Melo, C., Greaves, L., Saretzki, G., Fox, C., Lawless, C., Anderson, R., Hewitt, G., Pender, S.L., Fullard, N., Nelson, G., Mann, J., van de Sluis, B., Mann, D.A., von Zglinicki, T., 2014. Chronic inflammation induces telomere dysfunction and accelerates ageing in mice. Nat. Commun. 2, 4172. doi:10.1038/ncomms5172 Kang, S.-W., Kim, J., Shin, D.Y., 2016. Inhibition of senescence and promotion of the proliferation of chondrocytes from articular cartilage by CsA and FK506 involves inhibition of p38MAPK. Mech. Ageing Dev. 153, 7–13. doi:10.1016/j.mad.2015.12.002 Khoshnood, B., Dacklin, I., Grabbe, C., 2016. Urm1: an essential regulator of JNK signaling and oxidative stress in Drosophila melanogaster. Cell. Mol. Life Sci. 73, 1939–1954. doi:10.1007/s00018-015-2121-x Kim, C.D., Cha, J.D., Li, S., Cha, I.H., 2015. The mechanism of acacetin-induced apoptosis on oral squamous cell carcinoma. Arch. Oral Biol. 60, 1283–1298. doi:10.1016/j.archoralbio.2015.05.009 KIM, D.H., JUNG, W.-S., KIM, M.E., LEE, H.-W., YOUN, H.-Y., SEON, J.K., LEE, H.-N., LEE, J.S., 2014. Genistein inhibits pro-inflammatory cytokines in human mast cell activation through the inhibition of the ERK pathway. Int. J. Mol. Med. 34, 1669–1674. doi:10.3892/ijmm.2014.1956 Kim, H.-J., Ravichandran, K., Ozkok, A., Wang, Q., He, Z., Jani, A., Ljubanovic, D., Douglas, I.S., Edelstein, C.L., 2014. The water-soluble triptolide derivative PG490-88 protects against cisplatininduced acute kidney injury. J. Pharmacol. Exp. Ther. 349, 518–25. doi:10.1124/jpet.114.213769 Kim, H.J., Jung, K.J., Yu, B.P., Cho, C.G., Chung, H.Y., n.d. Influence of aging and calorie restriction on MAPKs activity in rat kidney. Exp. Gerontol. 37, 1041–53. Kim, K.-H., Park, B., Rhee, D.-K., Pyo, S., 2015. Acrylamide Induces Senescence in Macrophages through a Process Involving ATF3, ROS, p38/JNK, and a Telomerase-Independent Pathway. Chem. Res. Toxicol. 28, 71–86. doi:10.1021/tx500341z Kim, K.-N., Heo, S.-J., Yoon, W.-J., Kang, S.-M., Ahn, G., Yi, T.-H., Jeon, Y.-J., 2010. Fucoxanthin inhibits the inflammatory response by suppressing the activation of NF-κB and MAPKs in lipopolysaccharide-induced RAW 264.7 macrophages. Eur. J. Pharmacol. 649, 369–375. doi:10.1016/j.ejphar.2010.09.032 Kim, M., Park, K.Y., Lee, M.-K., Jin, T., Seo, S.J., 2016. Adiponectin Suppresses UVB-Induced Premature Senescence and hBD2 Overexpression in Human Keratinocytes. PLoS One 11, e0161247. doi:10.1371/journal.pone.0161247 Kim, M.C., Lee, H.J., Lim, B., Ha, K.-T., Kim, S.Y., So, I., Kim, B.J., 2014. Quercetin induces apoptosis by inhibiting MAPKs and TRPM7 channels in AGS cells. Int. J. Mol. Med. 33, 1657–63. doi:10.3892/ijmm.2014.1704 Kim, S.-J., Beak, S.-M., Park, S.-K., 2017. Supplementation with Triptolide Increases Resistance to Environmental Stressors and Lifespan in C. elegans. J. Food Sci. 82, 1484–1490. doi:10.1111/1750- 3841.13720 Kim, S.J., Li, M., Jeong, C.W., Bae, H.B., Kwak, S.H., Lee, S.H., Lee, H.J., Heo, B.H., Yook, K.B., Yoo, K.Y., 2014. Epigallocatechin-3-gallate, a green tea catechin, protects the heart against regional ischemia–reperfusion injuries through activation of RISK survival pathways in rats. Arch. Pharm. Res. 37, 1079–1085. doi:10.1007/s12272-013-0309-x Kim, S.M., Park, Y.J., Shin, M.S., Kim, H.R., Kim, M.J., Lee, S.H., Yun, S.P., Kwon, S.H., 2017. Acacetin inhibits neuronal cell death induced by 6-hydroxydopamine in cellular Parkinson’s disease model. Bioorganic Med. Chem. Lett. 27, 5207–5212. doi:10.1016/j.bmcl.2017.10.048 36 King, A.M., Van der Put, E., Blomberg, B.B., Riley, R.L., 2007. Accelerated Notch-dependent degradation of E47 proteins in aged B cell precursors is associated with increased ERK MAPK activation. J. Immunol. 178, 3521–9. Kovacheva, E.L., Hikim, A.P.S., Shen, R., Sinha, I., Sinha-Hikim, I., 2010. Testosterone supplementation reverses sarcopenia in aging through regulation of myostatin, c-Jun NH2-terminal kinase, Notch, and Akt signaling pathways. Endocrinology 151, 628–38. doi:10.1210/en.2009-1177 Kowshik, J., Giri, H., Kishore, T.K.K., Kesavan, R., Vankudavath, R.N., Reddy, G.B., Dixit, M., Nagini, S., 2014. Ellagic acid inhibits VEGF/VEGFR2, PI3K/Akt and MAPK signaling cascades in the hamster cheek pouch carcinogenesis model. Anticancer. Agents Med. Chem. 14, 1249–60. Kuroyanagi, G., Otsuka, T., Yamamoto, N., Matsushima-Nishiwaki, R., Kozawa, O., Tokuda, H., 2015. Resveratrol suppresses TGF-β-induced VEGF synthesis in osteoblasts: Inhibition of the p44/p42 MAPKs and SAPK/JNK pathways. Exp. Ther. Med. 9, 2303–2310. doi:10.3892/etm.2015.2389 Kwak, S.C., Lee, C., Kim, J.-Y., Oh, H.M., So, H.-S., Lee, M.S., Rho, M.C., Oh, J., 2013. Chlorogenic Acid Inhibits Osteoclast Differentiation and Bone Resorption by Down-Regulation of Receptor Activator of Nuclear Factor Kappa-B Ligand-Induced Nuclear Factor of Activated T Cells c1 Expression. Biol. Pharm. Bull 36, 1779–1786. doi:10.1074/jbc.M503347200 Kwong, J., Chen, M., Lv, D., Luo, N., Su, W., Xiang, R., Sun, P., 2013. Induction of p38 Expression Plays an Essential Role in Oncogenic ras-Induced Senescence. Mol. Cell. Biol. 33, 3780–3794. doi:10.1128/MCB.00784-13 Lai, C.-C., Huang, P.-H., Yang, A.-H., Chiang, S.-C., Tang, C.-Y., Tseng, K.-W., Huang, C.-H., 2016. Baicalein, a Component of Scutellaria baicalensis, Attenuates Kidney Injury Induced by Myocardial Ischemia and Reperfusion. Planta Med. 82, 181–189. doi:10.1055/s-0035-1558114 Lanna, A., Gomes, D.C.O., Muller-Durovic, B., McDonnell, T., Escors, D., Gilroy, D.W., Lee, J.H., Karin, M., Akbar, A.N., 2017. A sestrin-dependent Erk-Jnk-p38 MAPK activation complex inhibits immunity during aging. Nat. Immunol. 18, 354–363. doi:10.1038/ni.3665 Lanna, A., Henson, S.M., Escors, D., Akbar, A.N., 2014. The kinase p38 activated by the metabolic regulator AMPK and scaffold TAB1 drives the senescence of human T cells. Nat. Immunol. 15, 965–72. doi:10.1038/ni.2981 Lashmanova, E., Proshkina, E., Zhikrivetskaya, S., Shevchenko, O., Marusich, E., Leonov, S., Melerzanov, A., Zhavoronkov, A., Moskalev, A., 2015. Fucoxanthin increases lifespan of Drosophila melanogaster and Caenorhabditis elegans. Pharmacol. Res. 100, 228–241. doi:10.1016/j.phrs.2015.08.009 Lee, C.-H., Yoon, S.-J., Lee, S.-M., 2013. Chlorogenic acid attenuates high mobility group box 1 (HMGB1) and enhances host defense mechanisms in murine sepsis. Mol. Med. 18, 1437–48. doi:10.2119/molmed.2012.00279 Lee, E.B., Ahn, D., Kim, B.J., Lee, S.Y., Seo, H.W., Cha, Y.-S., Jeon, H., Eun, J.S., Cha, D.S., Kim, D.K., 2015. Genistein from Vigna angularis Extends Lifespan in Caenorhabditis elegans. Biomol. Ther. (Seoul). 23, 77–83. doi:10.4062/biomolther.2014.075 Lee, J.-J., Lee, J.-H., Ko, Y.-G., Hong, S.I., Lee, J.-S., 2010. Prevention of premature senescence requires JNK regulation of Bcl-2 and reactive oxygen species. Oncogene 29, 561–75. doi:10.1038/onc.2009.355 Lee, J., Kim, S., Namgung, H., Jo, Y.-H., Bao, C., Choi, H.-K., Auh, J.-H., Lee, H.J., 2014. Ellagic Acid Identified through Metabolomic Analysis Is an Active Metabolite in Strawberry (‘Seolhyang’) 37 Regulating Lipopolysaccharide-Induced Inflammation. J. Agric. Food Chem. 62, 3954–3962. doi:10.1021/jf4038503 Lee, J.H., Won, J.H., Choi, J.M., Cha, H.H., Jang, Y.J., Park, S., Kim, H.G., Kim, H.C., Kim, D.K., 2014. Protective Effect of Ellagic Acid on Concanavalin A-Induced Hepatitis via Toll-Like Receptor and Mitogen-Activated Protein Kinase/Nuclear Factor κB Signaling Pathways. J. Agric. Food Chem. 62, 10110–10117. doi:10.1021/jf503188c Lee, M., Yun, S., Lee, H., Yang, J., 2017. Quercetin Mitigates Inflammatory Responses Induced by Vascular Endothelial Growth Factor in Mouse Retinal Photoreceptor Cells through Suppression of Nuclear Factor Kappa B. Int. J. Mol. Sci. 18. doi:10.3390/ijms18112497 Lei, L., Zeng, Q., Lu, J., Ding, S., Xia, F., Kang, J., Tan, L., Gao, L., Kang, L., Cao, K., Zhou, J., Xiao, R., Chen, J., Huang, J., 2017. MALAT1 participates in ultraviolet B-induced photo-aging via regulation of the ERK/MAPK signaling pathway. Mol. Med. Rep. 15, 3977–3982. doi:10.3892/mmr.2017.6532 Li, Z., Li, J., Bu, X., Liu, X., Tankersley, C.G., Wang, C., Huang, K., 2011. Age-induced augmentation of p38 MAPK phosphorylation in mouse lung. Exp. Gerontol. 46, 694–702. doi:10.1016/j.exger.2011.04.005 Lian, W., Jia, H., Xu, L., Zhou, W., Kang, D., Liu, A., Du, G., 2017. Multi-Protection of DL0410 in Ameliorating Cognitive Defects in D-Galactose Induced Aging Mice. Front. Aging Neurosci. 9, 409. doi:10.3389/fnagi.2017.00409 Lin, F.-Y., Hsieh, Y.-H., Yang, S.-F., Chen, C.-T., Tang, C.-H., Chou, M.-Y., Chuang, Y.-T., Lin, C.-W., Chen, M.-K., 2015. Resveratrol suppresses TPA-induced matrix metalloproteinase-9 expression through the inhibition of MAPK pathways in oral cancer cells. J. Oral Pathol. Med. 44, 699–706. doi:10.1111/jop.12288 Lin, J., Yu, J., Zhao, J., Zhang, K., Zheng, J., Wang, J., Huang, C., Zhang, J., Yan, X., Gerwick, W.H., Wang, Q., Cui, W., He, S., 2017. Fucoxanthin, a Marine Carotenoid, Attenuates β-Amyloid Oligomer-Induced Neurotoxicity Possibly via Regulating the PI3K/Akt and the ERK Pathways in SH-SY5Y Cells. Oxid. Med. Cell. Longev. 2017, 6792543. doi:10.1155/2017/6792543 Lin, W.-H., Kuo, H.-H., Ho, L.-H., Tseng, M.-L., Siao, A.-C., Hung, C.-T., Jeng, K.-C., Hou, C.-W., 2015. Gardenia jasminoides extracts and gallic acid inhibit lipopolysaccharide-induced inflammation by suppression of JNK2/1 signaling pathways in BV-2 cells. Iran. J. Basic Med. Sci. 18, 555–62. Ling, Q., Meng, C., Chen, Q., Xing, D., 2014. Activated ERK/FOXM1 pathway by low-power laser irradiation inhibits UVB-induced senescence through down-regulating p21 expression. J. Cell. Physiol. 229, 108–16. doi:10.1002/jcp.24425 Liu, B., Jian, Z., Li, Q., Li, K., Wang, Z., Liu, L., Tang, L., Yi, X., Wang, H., Li, C., Gao, T., 2012. Baicalein protects Human melanocytes from H2O2-induced apoptosis via inhibiting mitochondriadependent caspase activation and the p38 MAPK pathway. Free Radic. Biol. Med. 53, 183–193. doi:10.1016/j.freeradbiomed.2012.04.015 Liu, C.-L., Lim, Y.-P., Hu, M.-L., 2013. Fucoxanthin enhances cisplatin-induced cytotoxicity via NFκB- mediated pathway and downregulates DNA repair gene expression in human hepatoma HepG2 cells. Mar. Drugs 11, 50–66. doi:10.3390/md11010050 Liu, M., Chen, J., Huang, Y., Ke, J., Li, L., Huang, D., Wu, W., 2015. Triptolide alleviates isoprenalineinduced cardiac remodeling in rats via TGF-β1/Smad3 and p38 MAPK signaling pathway. 38 Pharmazie 70, 244–50. Liu, Y., Li, J., Liu, Y., Wang, P., Jia, H., 2016. Inhibition of zymosan-induced cytokine and chemokine expression in human corneal fibroblasts by triptolide. Int. J. Ophthalmol. 9, 9–14. doi:10.18240/ijo.2016.01.02 López-Otín, C., Blasco, M.A., Partridge, L., Serrano, M., Kroemer, G., 2013. The Hallmarks of Aging. Cell 153, 1194–1217. doi:10.1016/j.cell.2013.05.039 Lorenzini, A., Tresini, M., Mawal-Dewan, M., Frisoni, L., Zhang, H., Allen, R.G., Sell, C., Cristofalo, V.J., n.d. Role of the Raf/MEK/ERK and the PI3K/Akt(PKB) pathways in fibroblast senescence. Exp. Gerontol. 37, 1149–56. Lu, Y., Jiang, F., Jiang, H., Wu, K., Zheng, X., Cai, Y., Katakowski, M., Chopp, M., To, S.-S.T., 2010. Gallic acid suppresses cell viability, proliferation, invasion and angiogenesis in human glioma cells. Eur. J. Pharmacol. 641, 102–7. doi:10.1016/j.ejphar.2010.05.043 Luo, X., Yu, Z., Deng, C., Zhang, J., Ren, G., Sun, A., Mani, S., Wang, Z., Dou, W., 2017. Baicalein ameliorates TNBS-induced colitis by suppressing TLR4/MyD88 signaling cascade and NLRP3 inflammasome activation in mice. Sci. Rep. 7, 16374. doi:10.1038/s41598-017-12562-6 Ma, J.-Q., Ding, J., Zhang, L., Liu, C.-M., 2014. Ursolic acid protects mouse liver against CCl4-induced oxidative stress and inflammation by the MAPK/NF-κB pathway. Environ. Toxicol. Pharmacol. 37, 975–83. doi:10.1016/j.etap.2014.03.011 Ma, X., Zhang, Y., Wang, Z., Shen, Y., Zhang, M., Nie, Q., Hou, Y., Bai, G., 2017. Ursolic Acid, a Natural Nutraceutical Agent, Targets Caspase3 and Alleviates Inflammation-Associated Downstream Signal Transduction. Mol. Nutr. Food Res. 61. doi:10.1002/mnfr.201700332 Ma, Y.-K., Chen, Y.-B., Li, P., 2018. Quercetin inhibits NTHi-triggered CXCR4 activation through suppressing IKKα/NF-κB and MAPK signaling pathways in otitis media. Int. J. Mol. Med. 42, 248– 258. doi:10.3892/ijmm.2018.3577 Marín, M., María Giner, R., Ríos, J.-L., Carmen Recio, M., 2013. Intestinal anti-inflammatory activity of ellagic acid in the acute and chronic dextrane sulfate sodium models of mice colitis. J. Ethnopharmacol. 150, 925–934. doi:10.1016/j.jep.2013.09.030 Markaki, M., Tavernarakis, N., 2013. Metabolic control by target of rapamycin and autophagy during ageing - a mini-review. Gerontology 59, 340–8. doi:10.1159/000348599 Marmisolle, I., Martínez, J., Liu, J., Mastrogiovanni, M., Fergusson, M.M., Rovira, I.I., Castro, L., Trostchansky, A., Moreno, M., Cao, L., Finkel, T., Quijano, C., 2017. Reciprocal regulation of acetyl-CoA carboxylase 1 and senescence in human fibroblasts involves oxidant mediated p38 MAPK activation. Arch. Biochem. Biophys. 613, 12–22. doi:10.1016/j.abb.2016.10.016 Martins, R., Lithgow, G.J., Link, W., 2016. Long live FOXO: unraveling the role of FOXO proteins in aging and longevity. Aging Cell 15, 196–207. doi:10.1111/acel.12427 McHugh, D., Gil, J., 2018. Senescence and aging: Causes, consequences, and therapeutic avenues. J. Cell Biol. 217, 65–77. doi:10.1083/jcb.201708092 Mei, X., Zhou, L., Zhang, T., Lu, B., Sheng, Y., Ji, L., 2018. Chlorogenic acid attenuates diabetic retinopathy by reducing VEGF expression and inhibiting VEGF-mediated retinal neoangiogenesis. Vascul. Pharmacol. 101, 29–37. doi:10.1016/j.vph.2017.11.002 Meng, L.-Q., Yang, F.-Y., Wang, M.-S., Shi, B.-K., Chen, D.-X., Chen, D., Zhou, Q., He, Q.-B., Ma, L.- 39 X., Cheng, W.-L., Xing, N.-Z., 2018. Quercetin protects against chronic prostatitis in rat model through NF-κB and MAPK signaling pathways. Prostate. doi:10.1002/pros.23536 Meng, Y., Zhang, Y., Ma, Z., Zhou, H., Ni, J., Liao, H., Tang, Q., 2017. Genistein attenuates pathological cardiac hypertrophy in vivo and in vitro. Herz. doi:10.1007/s00059-017-4635-5 Mi, Y., Qi, G., Fan, R., Qiao, Q., Sun, Y., Gao, Y., Liu, X., 2017. EGCG ameliorates high-fat– and highfructose–induced cognitive defects by regulating the IRS/AKT and ERK/CREB/BDNF signaling pathways in the CNS. FASEB J. 31, 4998–5011. doi:10.1096/fj.201700400RR Milman, S., Atzmon, G., Huffman, D.M., Wan, J., Crandall, J.P., Cohen, P., Barzilai, N., 2014. Low insulin-like growth factor-1 level predicts survival in humans with exceptional longevity. Aging Cell 13, 769–71. doi:10.1111/acel.12213 Ming, G.-F., Tang, Y.-J., Hu, K., Chen, Y., Huang, W.-H., Xiao, J., 2016. Visfatin attenuates the ox- LDL-induced senescence of endothelial progenitor cells by upregulating SIRT1 expression through the PI3K/Akt/ERK pathway. Int. J. Mol. Med. 38, 643–9. doi:10.3892/ijmm.2016.2633 Monti, D., Ostan, R., Borelli, V., Castellani, G., Franceschi, C., 2017. Inflammaging and human longevity in the omics era. Mech. Ageing Dev. 165, 129–138. doi:10.1016/j.mad.2016.12.008 Moskalev, A., Shaposhnikov, M., Zemskaya, N., Belyi, A., Dobrovolskaya, E., Patova, A., Guvatova, Z., Lukyanova, E., Snezhkina, A., Kudryavtseva, A., 2018. Transcriptome analysis reveals mechanisms of geroprotective effects of fucoxanthin in Drosophila. BMC Genomics 19, 77. doi:10.1186/s12864- 018-4471-x Negi, H., Shukla, A., Khan, F., Pandey, R., 2016. 3β-Hydroxy-urs-12-en-28-oic acid prolongs lifespan in C. elegans by modulating JNK-1. Biochem. Biophys. Res. Commun. 480, 539–543. doi:10.1016/j.bbrc.2016.10.073 Niu, Y., Na, L., Feng, R., Gong, L., Zhao, Y., Li, Q., Li, Y., Sun, C., 2013. The phytochemical, EGCG, extends lifespan by reducing liver and kidney function damage and improving age-associated inflammation and oxidative stress in healthy rats. Aging Cell 12, 1041–1049. doi:10.1111/acel.12133 Noh, E.-M., Park, Y.-J., Kim, J.-M., Kim, M.-S., Kim, H.-R., Song, H.-K., Hong, O.-Y., So, H.-S., Yang, S.-H., Kim, J.-S., Park, S.H., Youn, H.-J., You, Y.-O., Choi, K.-B., Kwon, K.-B., Lee, Y.-R., 2015. Fisetin regulates TPA-induced breast cell invasion by suppressing matrix metalloproteinase-9 activation via the PKC/ROS/MAPK pathways. Eur. J. Pharmacol. 764, 79–86. doi:10.1016/j.ejphar.2015.06.038 Oh, J., Lee, Y.D., Wagers, A.J., 2014. Stem cell aging: mechanisms, regulators and therapeutic opportunities. Nat. Med. 20, 870–880. doi:10.1038/nm.3651 Oh, S.W., Mukhopadhyay, A., Svrzikapa, N., Jiang, F., Davis, R.J., Tissenbaum, H.A., 2005. JNK regulates lifespan in Caenorhabditis elegans by modulating nuclear translocation of forkhead transcription factor/DAF-16. Proc. Natl. Acad. Sci. 102, 4494–4499. doi:10.1073/pnas.0500749102 Okuyama, T., Inoue, H., Ookuma, S., Satoh, T., Kano, K., Honjoh, S., Hisamoto, N., Matsumoto, K., Nishida, E., 2010. The ERK-MAPK pathway regulates longevity through SKN-1 and insulin-like signaling in Caenorhabditis elegans. J. Biol. Chem. 285, 30274–81. doi:10.1074/jbc.M110.146274 Pal, H.C., Sharma, S., Strickland, L.R., Katiyar, S.K., Ballestas, M.E., Athar, M., Elmets, C.A., Afaq, F., 2014. Fisetin inhibits human melanoma cell invasion through promotion of mesenchymal to epithelial transition and by targeting MAPK and NFκB signaling pathways. PLoS One 9, e86338. doi:10.1371/journal.pone.0086338 40 Pan, W., Yu, H., Huang, S., Zhu, P., 2016. Resveratrol Protects against TNF-α-Induced Injury in Human Umbilical Endothelial Cells through Promoting Sirtuin-1-Induced Repression of NF-KB and p38 MAPK. PLoS One 11, e0147034. doi:10.1371/journal.pone.0147034 Pang, J.-H.S., Yen, J.-H., Wu, H.-T., Huang, S.-T., 2017. Gallic Acid Inhibited Matrix Invasion and AP- 1/ETS-1-Mediated MMP-1 Transcription in Human Nasopharyngeal Carcinoma Cells. Int. J. Mol. Sci. 18. doi:10.3390/ijms18071354 Pangestuti, R., Vo, T.-S., Ngo, D.-H., Kim, S.-K., 2013. Fucoxanthin Ameliorates Inflammation and Oxidative Reponses in Microglia. J. Agric. Food Chem. 61, 3876–3883. doi:10.1021/jf400015k Pant, A., Saikia, S.K., Shukla, V., Asthana, J., Akhoon, B.A., Pandey, R., Pandey, R., 2014. Betacaryophyllene modulates expression of stress response genes and mediates longevity in Caenorhabditis elegans. Exp. Gerontol. 57, 81–95. doi:10.1016/j.exger.2014.05.007 Park, K.-R., Nam, D., Yun, H.-M., Lee, S.-G., Jang, H.-J., Sethi, G., Cho, S.K., Ahn, K.S., 2011. β- Caryophyllene oxide inhibits growth and induces apoptosis through the suppression of PI3K/AKT/mTOR/S6K1 pathways and ROS-mediated MAPKs activation. Cancer Lett. 312, 178– 188. doi:10.1016/j.canlet.2011.08.001 Park, W.H., Kim, S.H., 2013. MAPK inhibitors augment gallic acid-induced A549 lung cancer cell death through the enhancement of glutathione depletion. Oncol. Rep. 30, 513–519. doi:10.3892/or.2013.2447 Patwardhan, R.S., Sharma, D., Checker, R., Sandur, S.K., 2014. Mitigation of radiation-induced hematopoietic injury via regulation of cellular MAPK/phosphatase levels and increasing hematopoietic stem cells. Free Radic. Biol. Med. 68, 52–64. doi:10.1016/j.freeradbiomed.2013.11.004 Pauli, J.R., Ropelle, E.R., Cintra, D.E., De Souza, C.T., da Silva, A.S.R., Moraes, J.C., Prada, P.O., de Almeida Leme, J.A.C., Luciano, E., Velloso, L.A., Carvalheira, J.B.C., Saad, M.J.A., 2010. Acute exercise reverses aged-induced impairments in insulin signaling in rodent skeletal muscle. Mech. Ageing Dev. 131, 323–9. doi:10.1016/j.mad.2010.03.004 Peng, Z., Gong, X., Yang, Y., Huang, L., Zhang, Q., Zhang, P., Wan, R., Zhang, B., 2017. Hepatoprotective effect of quercetin against LPS/ d -GalN induced acute liver injury in mice by inhibiting the IKK/NF-κB and MAPK signal pathways. Int. Immunopharmacol. 52, 281–289. doi:10.1016/j.intimp.2017.09.022 Peti, W., Page, R., 2013. Molecular basis of MAP kinase regulation. Protein Sci. 22, 1698–1710. doi:10.1002/pro.2374 Pietsch, K., Saul, N., Menzel, R., Stürzenbaum, S.R., Steinberg, C.E.W., 2009. Quercetin mediated lifespan extension in Caenorhabditis elegans is modulated by age-1, daf-2, sek-1 and unc-43. Biogerontology 10, 565–78. doi:10.1007/s10522-008-9199-6 Plotnikov, A., Zehorai, E., Procaccia, S., Seger, R., 2011. The MAPK cascades: Signaling components, nuclear roles and mechanisms of nuclear translocation. Biochim. Biophys. Acta - Mol. Cell Res. 1813, 1619–1633. doi:10.1016/j.bbamcr.2010.12.012 Proshkina, E., Lashmanova, E., Dobrovolskaya, E., Zemskaya, N., Kudryavtseva, A., Shaposhnikov, M., Moskalev, A., 2016. Geroprotective and Radioprotective Activity of Quercetin, (-)-Epicatechin, and Ibuprofen in Drosophila melanogaster. Front. Pharmacol. 7, 505. doi:10.3389/fphar.2016.00505 Qian, Y., Cao, L., Guan, T., Chen, L., Xin, H., Li, Y., Zheng, R., Yu, D., 2015. Protection by genistein on cortical neurons against oxidative stress injury via inhibition of NF-kappaB, JNK and ERK 47 doi:10.1038/onc.2013.477 Zhu, B., Ferry, C.H., Markell, L.K., Blazanin, N., Glick, A.B., Gonzalez, F.J., Peters, J.M., 2014. The Nuclear Receptor Peroxisome Proliferator-activated Receptor-β/δ (PPARβ/δ) Promotes Oncogeneinduced Cellular Senescence through Repression of Endoplasmic Reticulum Stress. J. Biol. Chem. 289, 20102–20119. doi:10.1074/jbc.M114.551069 Zou, Y., Wei, H.K., Xiang, Q.-H., Wang, J., Zhou, Y.-F., Peng, J., 2016. Protective effect of quercetin on pig intestinal integrity after transport stress is associated with regulation oxidative status and inflammation. J. Vet. Med. Sci. 78, 1487–1494. doi:10.1292/jvms.16-0090