review article biological significance of urolithins,...

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 270418 15 pageshttpdxdoiorg1011552013270418

Review ArticleBiological Significance of Urolithins the Gut Microbial EllagicAcid-Derived Metabolites The Evidence So Far

Juan Carlos Espiacuten Mar Larrosa Mariacutea Teresa Garciacutea-Conesaand Francisco Tomaacutes-BarberaacutenResearch Group on Quality Safety and Bioactivity of Plant Foods Department of Food Science and TechnologyCEBAS-CSIC Campus de Espinardo PO Box 164 30100 Murcia Spain

Correspondence should be addressed to Francisco Tomas-Barberan fatomascebascsices

Received 4 January 2013 Revised 15 April 2013 Accepted 17 April 2013

Academic Editor David Heber

Copyright copy 2013 Juan Carlos Espın et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The health benefits attributed to pomegranate have been associated with its high content in polyphenols particularly ellagitanninsThis is also the case for other ellagitannin-containing fruits and nuts including strawberry raspberry blackberry walnutsand muscadine grapes The bioavailability of ellagitannins and ellagic acid is however very low These molecules sufferextensive metabolism by the gut microbiota to produce urolithins that are much better absorbed Urolithins circulate inplasma as glucuronide and sulfate conjugates at concentrations in the range of 02ndash20 120583M It is therefore conceivable thatthe health effects of ellagitannin-containing products can be associated with these gut-produced urolithins and thus theevaluation of the biological effects of these metabolites is essential Recent research mostly based on in vitro testing hasshown preliminary evidence of the anti-inflammatory anticarcinogenic antiglycative antioxidant and antimicrobial effects ofurolithins supporting their potential contribution to the health effects attributed to pomegranate and ellagitannin-rich foodsThe number of in vivo studies is still limited but they show preventive effects of urolithins on gut and systemic inflammationthat encourage further research Both in vivo and mechanistic studies are necessary to clarify the health effects of thesemetabolites Attention should be paid when designing these mechanistic studies in order to use the physiologically relevantmetabolites (urolithins in gut models and their conjugated derivatives in systemic models) at concentrations that can be reachedin vivo

1 Introduction

The health effects of pomegranate and pomegranate juicehave been associated withthe high content in antioxidantpolyphenols [1] and particularly in ellagitannins (punicala-gins) This is also the case for many other ellagitannin-containing fruits and nuts such as strawberry raspberryblackberry walnuts and muscadine grapes [2 3]

Both ellagic acid and ellagitannins have shown relevantbiological effects in animal models and human studieswhich suggests potential preventive effects against chronicdiseases such as cancer diabetes cardiovascular diseases andneurodegenerative diseases [4] These effects are associated

with a multitarget action that involves antioxidant anti-inflammatory and anticarcinogenic effects [2 5] It is wellestablished that ellagitannins and ellagic acid absorptionis very low and that the unabsorbed compounds are fur-ther metabolized to urolithins by the gut microbiota inthe colon [6ndash10] This poor bioavailability and the extensivegut catabolism suggest that urolithins rather than ellagitan-nins or ellagic acid may be the actual bioactive molecules[7 11]

Urolithins are dibenzopyran-6-one derivatives with dif-ferent hydroxyl substitutions Chemically they can be con-sidered a combination of coumarin and isocoumarin (benzo-coumarins) (Figure 1)They are produced from ellagic acid bythe gut microbiota through the loss of one of the two lactones

2 Evidence-Based Complementary and Alternative Medicine

HO

HO

OO

OO

OO

OH

OH

Urolithin A

Coumarin Isocoumarin

Figure 1 Chemical structure of urolithin A a benzocoumarin

present in ellagic acid (lactonasedecarboxylase activity) andby successive removals of hydroxyls (dehydroxylase activi-ties) [12] (Figure 2)

Urolithins are not commonmolecules in nature but theyhave been reported in plants rich in ellagitannins as it is thecase of Tamarix nilotica flowers [13] and Punica granatumleaves that contain urolithin M-5 (348910-pentahydroxy-dibenzo[bd]pyran-6-one) [14] They have also been foundin some herbivore-derived products such as the castoreumproduced by beavers [15] a product of interest for theperfume industry and thePteropi faeces (faeces of the squirrelTrogopterus xanthipes) that are used in traditional Chinesemedicine [16] Urolithins are also relevant constituents ofshilajit a soil-derived medicinal product used in Ayurvedicmedicine [17]

The name urolithin was first given to two metabolitesisolated from the renal calculus of sheep (Trifolium sub-terraneum has been reported as the cause of clover stoneand might be a relevant source of ellagitannins) that werenamed urolithin A and urolithin B [18] These two moleculescoincided with pigments I and II previously described fromcastoreum and beaver glands [15]

The intake of large amounts of ellagitannins and particu-larly of punicalagin from Terminalia oblongata by cattle wasassociated with hepatotoxicity and nephrotoxicity [19] How-ever the subchronic oral administration of large amountsof pomegranate ellagitannins was not toxic to rats wherethese compounds were found to be extensively metabolizedto urolithins [20]

Since urolithins are ellagitannin-derived catabolites thatcan be absorbed and reach different tissues in the body theyhave been suggested as the molecules potentially respon-sible for the biological effects observed as a consequenceof the consumption of pomegranate or other ellagitannin-containing foods [3 7 9 11] Here we review the state of theart in urolithin metabolites production by gut microbiotatheir absorption tissue distribution and pharmacokineticsthe cell andmolecular mechanisms for their biological effectsreported so far using different in vitromodels and the in vivoevidence in animals and humans

2 Urolithins Chemical NatureDetection and Identification

Urolithins constitute a whole metabolic family producedby the opening and decarboxylation of one of the lactonerings of ellagic acid and the sequential removal of hydroxylsfrom different positions (Figure 2) After decarboxylation thefirst metabolite is urolithin M-5 (pentahydroxy-urolithin)and from this several tetrahydroxy-urolithin isomers areproduced by removal of one hydroxyl group from differentpositions (urolithin D urolithinM-6) Trihydroxy-urolithins(urolithin C urolithin M-7) were then produced after theremoval of a second hydroxyl and dihydrox- urolithins(urolithin A and isourolithin A) after the removal of athird one Monohydroxy-urolithin (urolithin B) was alsodetected particularly in those cases in which isourolithin Awas produced (Figure 2) Further degradation of urolithinsto remove the second lactone ring has not been reportedso far although it should not be discarded Urolithins showcharacteristic UV spectra that can be used for the identi-fication of different hydroxyl-substitution patterns on theurolithin nucleus as well as the further conjugation withmethyl glucuronic acid or sulfate [21] The UV spectra ofurolithins in methanol exhibit two major absorption bandsin the region of 240ndash400 nm (Figure 4) These are referred toas band I (300ndash380 nm) and band II (240ndash280 nm) In manycases an additional band III (between 280 and 300 nm) is alsoobserved Unfortunately bands I and II cannot be associatedwith a specific ring of the urolithin nucleus [21] Two groupsof urolithinUV spectra were distinguished after a study of theplots of the different spectra urolithins with a hydroxyl in the9-position and those without hydroxylation in the 9-position(Figure 4) The occurrence of the hydroxyl at position 9 as isthe case of isourolithin A produces a hypsochromic shift inband I and an increase in the absorption of band II displayinga maximum around 256 nm which is the main absorptionband of the spectrum of urolithins with a hydroxyl in the 9-position Conjugation with glucuronic acid or sulfate also hasmeasurable effects on the UV spectrum and could be used asa diagnostic method for urolithin metabolites identification

3 Bioavailability Metabolism and TissueDistribution of Pomegranate Ellagitanninsand Their Metabolites

Due to the potential health effects and the expectationraised by the high antioxidant activity of pomegranate juicesthat was thought to be due to punicalagins punicalins andother ellagitannins [1 22] a sensible step in the researchwas the determination of the absorption and metabolism ofthese compounds in animals and humans The first studytesting the bioavailability of pomegranate ellagitannins wascarried out on rats and this showed that after the intake oflarge amounts of pomegranate husk ellagitannins the mainmetabolites detected in plasma and urine were urolithinsA B and C (Figure 2) and smaller amounts of ellagicacid-dimethyl ether glucuronide [6] Interestingly a smallamount of punicalagin was also detected in plasma and urine

Evidence-Based Complementary and Alternative Medicine 3

O

OO

O

OO

COOH

OO

OO

OO

OO O

O

OO

OO

OOO

O

Ellagitannins

Ellagic acid

Urolithin M-5

Urolithin D

Urolithin M-7Urolithin C

Urolithin A

Urolithin B

Intermediate

Urolithin M-6

Isourolithin A

Isourolithin B

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HOHO

HO

HO

HO

HO

HO

OH

OHOH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OHOH

OH

Figure 2 Gut microbiota metabolism of ellagitannins and ellagic acid

particularly after a long period of intake but this has notbeen confirmed in further studies in humans inwhich dietaryrelevant amounts of pomegranate ellagitannins were supplied[7]

Urolithin A was produced from ellagic acid punicalaginand an ellagitannin-rich walnut extract by fecal microbiotafrom six volunteers demonstrating for the first time theproduction of urolithins by human gut microbiota [12] Inaddition a large interindividual variability was observed inthis ex vivo experiment [12] in agreement with the resultsobserved in vivo [7] suggesting that differences in the micro-biota composition affect urolithin production and therefore

the potential health effects after consumption of ellagitannin-rich foods [12] No correlation between urolithin productionfrom ellagitannins and equol production from isoflavoneswas observed [12] and this indicated that different bacterialstrains are involved in the gut metabolism of isoflavones andellagitannins

The Iberian pig was used as a model to study urolithinproduction from ellagitannins This particular pig feeds onoak acorns that are very rich in ellagitannins and thismodel was used to evaluate ellagitannin metabolism andtissue distribution [23] and to help in understanding theellagitannin metabolism in humans This study showed that


OO

Urolithin A(URO-A)

OO

Ellagitannins

Ellagic acid

O

OO

O

OO

Urolithin B(URO-B)

OO

Esophagus

Tetrahydroxy-benzopyran-6-one(URO-M6)

Trihydroxy-benzopyran-6-one(URO-C)

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

OH

OH

Bact

eria

l met

abol

ism

Figure 3 Sequential production of urolithins in the gut

different urolithins were produced in the gut starting withtetrahydroxy-urolithin through the removal of one of thelactone rings of ellagic acid and by sequential removal ofhydroxyls to end with urolithins A and B (Figure 3) Theanalysis of plasma and urine samples showed that urolithinA glucuronide and sulfate were the main metabolites withurolithinC andB glucuronides and sulfates asminormetabo-lites Ellagic acid dimethyl ether glucuronide was also asignificantmetaboliteThe analysis of the gall bladder and bileshowed that tetrahydroxy-urolithin was absorbed in the firstportion of the gut and was detected in the liver where it wasconjugated and excreted with the bile to the small intestineAn entero-hepatic recirculation was clearly revealed in thisanimal model study [23] Regarding the tissue distributionurolithin metabolites only accumulated in the urine bladderand the gall bladder where they reached high concentrationsbut they did not accumulate in any of the analyzed tissues(muscle adipose tissue kidney liver heart etc) [23]

The occurrence of low concentrations or urolithin met-abolites in mouse prostate gland after pomegranate ellagi-tannins intake [24] and in human prostate after the intakeof pomegranate juice and walnuts [25] has been reportedThis last study is the only one available on the evaluation of

the occurrence of ellagitannin metabolites in human tissues(prostate biopsies) and the occurrence of urolithin A glu-curonide (2 ngg tissue) and traces of urolithin B glucuronideand ellagic acid dimethyl ether was reported [25]

There are no studies on the pharmacokinetics and tissuedistribution of urolithins after their direct intake in humansThere is however indirect evidence of both aspects afterthe intake of ellagitannins or ellagic acid-rich food productsIn the case of pomegranates or derived beverages it iswell established that ellagitannins are not absorbed whendietary doses are ingested and that the released ellagic acidis absorbed in the first part of the gastrointestinal tractand is detected as such in plasma with 119862max concentrationsaround 100 nM with 119879max 1 hour after the intake [10 26]Ellagic acid conjugates have also been detected and theseincludemethyl ether and glucuronyl and sulphate conjugatesThe most common metabolite found in urine and plasmais ellagic acid dimethyl ether glucuronide which involvesthe methylation by COMT and then the glucuronidationby glucuronyl-transferase [3 7] The peak plasma levels ofurolithin A were 14ndash25 120583M depending on the volunteersThese metabolites started to appear in plasma 6ndash8 hoursafter the intake which confirms urolithin production in the


OO

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Band IBand IIIBand II

120582 (nm)

Urolithin A

HO OH

(a)

OO

256

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mAU

0

500

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302329


120582 (nm)Isourolithin A

HO

OH

(b)

Figure 4 UV characteristic spectra of urolithins

colon [7] and persisted in urine and plasma 48ndash72 hoursafter the ellagitannins intake showing an active enterohepaticrecirculation [3 7 9 12]

Seeram et al [24] described the pharmacokinetics andtissue distribution of urolithins in mice after oral or intra-peritoneal administration of chemically synthesized urolithinA (human equivalent dose HED sim85mg for a 70 kg person)Urolithin A peaked in plasma after 2 h and reached thehighest concentrations in the prostate followed by the smallintestine and colon with a peak at 4 h [24] Urolithin Aurolithin A sulfate and methyl-urolithin A were mainlydetected in the prostate gland whereas urolithin A glu-curonide was primarily detected in liver and kidney tissuesOther studies with animal models including rats and pigsconfirmed the urolithin production in the colon and thesequential loss of hydroxyls when the metabolites advance inthe intestinal tract [3 12]

Gonzalez-Sarrıas et al [25] reported the occurrence ofurolithin metabolites and ellagic acid dimethyl ether glu-curonide in prostate biopsies after the intake of pomegranatejuice This is the only study carried out so far to evaluate theoccurrence of urolithin metabolites in human tissues Theconcentration detected in the prostate gland was howeverlow although higher concentrations should not be discardedas suggested by a parallel study carried out in rats [25] andalso in agreement with the results obtained by Seeram et al[24]

4 Production of Urolithins byDifferent Animals

The production of urolithins from ellagitannins has beenreported in different animals (Table 1) Those animals thatfeed on bark and wood as is the case of beavers and squirrelsproduce urolithins in their gut and they are present in theirfeces and in excretionssecretions as is the case of castoreum

In addition ruminants that feed on ellagitannin-rich fod-ders (oak leaves Trifolium subterraneum etc) also produceurolithins that circulate in plasma as conjugated derivativesand are excreted in urine and feces The production ofurolithins in the rumen of cattle has been demonstrated andisourolithin A and urolithin B were the main metabolitesobserved while urolithin A seems to be mainly producedin the intestine [31] The kinetics of the production of thedifferent metabolites suggests that urolithin B productionfrom isourolithin A is more favored than its production fromurolithin A that seems to be an end product The occurrenceof urolithins in milk seems likely although it has not beendemonstrated so far

In monogastric animals urolithins are produced in ratmouse and pig and in all these cases the main metabolitepresent in feces urine and plasma is urolithin A and itsglucuronide and sulfate conjugates Urolithins C and B areless frequent but they have also been detected together with


Table 1 Production of urolithins as ellagitannin metabolites in different animals and humans

Animal production Ellagitannin source Metabolites ReferencesRat (Rattus norvegicus) Pomegranate husk Uro A Uro B Uro C [6 20]Rat (Rattus norvegicus) Ellagic acid Uro A [27]Rat (Rattus norvegicus) Oak-flavored milk Uro A Uro B Uro C [28]Rat (Rattus norvegicus) Pomegranate extract Uro A [29]

Rat (Rattus norvegicus) Geraniin (from Geranium thunbergii) Uro A Uro M-6 Uro M-7Uro M5 [30]

Mouse (Mus musculus) Pomegranate extract Uro A [24]Mouse (Mus musculus) Pomegranate husk Uro A [21]

Beaver (Castor canadensis) Wood Uro A Uro B [15][21]

Complex-toothed squirrel (Trogopterusxanthipes) Unknown Uro A [16 21]

Sheep (Ovis aries) Trifolium subterraneum Uro A Uro B [18]Sheep (Ovis aries) Quebracho Uro A lowast

Cattle (Bos primigenius) Young oak leaves Uro A IsoUro A Uro B [31]Pig (Sus scrofa domesticus) Acorns Uro A Uro C Uro D Uro B [23]

Humans (Homo sapiens) Pomegranate juice Uro A Uro C Isouro AUro B

[7 9 2532]

Humans (Homo sapiens) Pomegranate extract Uro A UroB Uro C [10]

Humans (Homo sapiens) Walnuts Uro A Uro B Uro C [8][25]

Humans (Homo sapiens) Strawberry Uro A Isouro A Uro BUro C [8 33]

Humans (Homo sapiens) Raspberry Uro A IsoUro A Uro B [8 34]Humans (Homo sapiens) Blackberry Uro A Uro C lowast

Humans (Homo sapiens) Cloudberry Uro A lowast

Humans (Homo sapiens) Oak-aged red wine Uro A [8]Humans (Homo sapiens) Tea Uro A [35]Humans (Homo sapiens) Nuts Uro A Isouro A Uro B [36]Uro A (urolithin A) Uro B (urolithin B) Uro C (urolithin C) Uro D (urolithin D) Isouro A (isourolithin A) Uro M-5 (urolithin M-5) Uro M-6 (urolithinM-6) and Uro M-7 (urolithin M-7) lowastTomas-Barberan et al unpublished results

small amounts of isourolithin A (Table 1) The same behavioris observed in humans after the intake of pomegranateswalnuts tea muscadine grapes strawberries raspberriesblackberries cloudberries oak acorns and oak-aged redwine Urolithins will also be eventually produced after theintake of all ellagitannin-containing foods and medicinalplants as is the case of camu camu (Myrciaria dubia) arcticbramble rose hip sea buckthorn cranberry Geranium andoak-aged spirits (whisky etc)

The studies to evaluate urolithin production in otheranimals including birds and insects are very limited [21]but the evidence so far indicates that they do not produceurolithins from ellagitannins In insects ellagitannins arehydrolyzed to release ellagic acid which is detected in fecesThis was shown in the acorn beetle (Thorectes lusitanicus)[21] Indirect evidence shows that honeybees harvestingnectar containing ellagitannins hydrolyze them to releaseellagic acid during honey maturation [37] but no urolithinsare detected suggesting that honeybee microbiota is not able

to metabolize ellagic acid to produce urolithins In termitesfeeding on wood ellagitannins are hydrolyzed to ellagic acidand then hydroxyls are removed to produce nasutins andit seems that termite microbiota does not have the abilityof removing the lactone ring of ellagic acid to produceurolithins [21] It has been reported that the hemolymph ofsome Australian termites of the genus Nasutitermes containsnasutin A that is also present in feces but no urolithin wasdetected [38]

In birds the only study available was done with green-finches (Carduelis chloris) that were fed for 2 weeks withblackberries Feces were collected and analyzed and onlyellagic acid was detected [21]

These results show that urolithins are generally producedby mammals after the intake of ellagitannins There ishowever interindividual variability that has been suggestedto be associated with different gut microbiota compositionand thismeans that the health effects observed after the intakeof pomegranates and other ellagitannin-containing food can


Table 2 Biological effects of urolithins assayed on human cell lines

Test compound Activity Test model Doseduration Effect Ref

UA UB UC UD AntioxidantPMADCFH-DAHL-60 cells

004ndash137 120583M 05 h Inhibit cellular injury caused byROS [39]

UA UB Antioxidant DMNQSK-N-MC cells 01ndash20 120583M 05 h Increase cell survival [40]

UA UB EstrogenicAnti-estrogenic MCF-7 cells 01ndash40 120583M 7 d

Induction of cell proliferation(estrogenic)inhibition of cellproliferation in the presence ofestradiol (anti-estrogenic)

[41]

UA UB Antimalarial THP-1 stimulated withhaemozoin or TNF-120572 25 120583M 48 h Inhibit the release and expression

of MMP-9 [42]

UA UB Anti-inflammatory CCD18-CoTHP-1stimulated with TNF-120572 5ndash40 120583M 48 h

Decrease fibroblasts migrationand monocyte adhesion tofibroblasts

[43]

UA UB Anti-inflammatory CCD18-Co stimulated withIL-1120573 01ndash40 120583M 18 h Inhibit PGE2 production and

mPGES-1 and COX-2 expression [44]

UA UB UAG UBG Anti-inflammatory HAOECTHP-1 stimulatedwith TNF-120572 1ndash20 120583M 4ndash24 h Inhibit monocyte adhesion and

endothelial cell migration [45]

UA UB UC UD Anticancer 22Rv1 EROD assay 675ndash50 120583M 05 24 h Inhibit CYP1B1 and CYP1A1activity [46]

UA UB Anticancer Caco-2 cells 40 120583M 24ndash96 h Cell cycle arrest in S and G2Mphases [47]

UA Anticancer HEK T293 cells Wntluciferase assay

02ndash200 120583gmL48 h Wnt pathway inhibition [48]

UA UB UC UD Anticancer HT-29 cells 25ndash70 120583M Apoptosis induction and CYP1activity inhibition [49]

Caco-2 human colon carcinoma cell line CCD18-Co human colon fibroblast cell line CYP1A1 cytochrome P450 family 1 member A1 CYP1B1 cytochromeP450 family 1 member B1 DCFH-DA 2101584071015840-dichlorodihydrofluorescein diacetate DMNQ 23-dimethoxy-14-naphthoquinone EROD ethoxyresorufin-O-deethylase assay HAOEC human aortic endothelial cell line HEK T293 human embryonic kidney cell line HL-60 promyelocytic leukaemia cells HT-29human colon carcinoma cell line MCF-7 human breast cancer cell line PGE2 prostaglandin E2 PMA phorbol 12-myristate-13-acetate SK-N-MC humanneuroblastoma cells THP-1 human acute monocytic leukaemia cell line UA urolithin A UAG urolithin A glucuronide UB urolithin B UBG urolithin Bglucuronide UC urolithin C UD urolithin D 22Rv1 human prostate carcinoma cell line

be modulated by the occurrence of specific microbiota toproduce urolithins [50 51]

5 Biological Activity of Urolithins

The bioavailability and metabolism studies clearly indicatewhich metabolites should be tested and at what concentra-tions in the mechanistic studies to understand the biologicalactivity of pomegranate ellagitannins A combination ofpunicalagin ellagic acid and urolithins should be testedin those studies using models of gastrointestinal tract cellswhere they can reach concentrations around several hundred120583M (for punicalagin and ellagic acid) and tens 120583M forurolithins

The health effects attributed to urolithins based on studiescarried out in vitro are numerous and diverse from anti-malarial properties or topoisomerase inhibitors to quenchersof bacterial quorum sensing In general the number ofpublications regarding each biological activity is still verylimited although these studies are generally carried out using

a suitable physiological concentration range in the order ofsubmicromolar-low micromolar concentrations similar tothose that urolithins can reach in the gut and plasma afterthe ingestion of ellagitannin-rich foods (Table 2) Fromall thestudies carried out in vitro it is important to highlight a recentstudy inwhich the biological effects of urolithin glucuronidesthe major circulating metabolites in plasma after ingestionof ellagitannin-rich foods were explored in human aorticendothelial cells [45] This type of study is still very scarcealthough highly desirable when studying the mechanismof action of polyphenols or their metabolites outside thegastrointestinal tract as polyphenols and their gutmicrobiotametabolites are glucuronidated by Phase II enzymes afterabsorption and they are found in this conjugated form in thecirculatory system and peripheral tissues

51 Antioxidant Activity Given the high antioxidant proper-ties of ellagitannin-rich foods as is the case of pomegranate[1] one of the first activities that had been explored for


urolithins was their antioxidant capacity The first availablestudy reported that urolithin A had an antioxidant capacity42-times lower than that of its precursor punicalagin when itwas tested in the DPPH assay and 3500 times lower when itwas tested in theABTS assay [7]These results were consistentwith the concentrations of urolithins necessary to reach anIC50

in several antioxidant assays (DPPH xanthine-XODand PMS-NADH) that were above 100 120583M [29] Howeverusing the ORAC assay all urolithins tested exhibited potentantioxidant properties compared with those of ascorbic acidurolithin A being the most potent among them [29] In asimilar study in which the antioxidant activity of a largenumber of polyphenols and their metabolites producedin vivo was compared by the ORAC method the antioxidantactivity of urolithin A was one of the most powerful onlybeaten by some proanthocyanidin oligomers catechin epi-catechin and 34-dihydroxyphenyl acetic acid [52] Using acell-based method in which the transport through the cellmembrane was considered Bialonska et al [39] determinedthat urolithinC showed the highest antioxidant power (IC

50=

016 120583M) whereas urolithin A showed an IC50of 136 120583M still

in the range of the plasma concentrations achievable in vivobut in a lesser degree than its precursor ellagic acid (11120583M)or vitamin C (19 120583M) [39] In neuronal cells in which anoxidative stress was induced urolithin B (05ndash20 120583M) andurolithin A (10 120583M) exhibited a protective effect increasingcell survival [40] In general it is assumable that urolithinshave higher antioxidant capacity than was originally thoughtbut the antioxidant capacity depends on the measurementmethod and needs to be studied inmore detail andwith otherin vivomethods given the importance of free radicals inmanydiseases

52 Estrogenic Modulators In the last few years therehas been an increasing interest in the study of the estro-genicantiestrogenic activity of plant-derived compounds(phytoestrogens) due to their potential benefits as part of thediet that is regulation of cholesterol levels or maintenanceof the bone density after menopause Many polyphenols(isoflavones flavanones estilbenes etc) show phytoestro-genic effects In some cases dietary polyphenols may bethe precursors of the so-called enterophytoestrogens thatare produced by the colon microbiota by catabolism ofthe original phenolics The potential activity of urolithinsas enterophytoestrogens has recently been investigated [41]In this work structure-activity studies revealed that theurolithins A and B had specific molecular characteristics thatmade these molecules potentially able to bind with the 120572-and 120573-estrogenic receptors (ER) [41] The ER CompetitiveBinding Assays showed that both urolithins had an affinityfor the ER120572 and ER120573 receptors that urolithin A bound moreeffectively than urolithin B and that the affinitywas higher forthe ER120572 than for the ER120573 receptor Using a proliferation assaywith cells sensitive to estrogens (E-screen) urolithins A andB exhibited estrogenic activity (in the absence of estradiol)and antiestrogenic activity (in the presence of estradiol) in asimilar fashion to other known phytoestrogens [41]

53 Anti-Inflammatory-Related Activities

531 Antimalarial Activity In an attempt to clarify whetherthe properties of the sun-dried rind of immature Punicagranatum that is used as an anti-malarial herbal rem-edy are at least in part due to urolithins their activityinhibiting the MMP-9 enzyme (directly involved in thepathogenesis of malaria) was tested Urolithins A and B atconcentrations of 25120583M inhibited the release of MMP-9 andits mRNA expression in hemozoin and TNF-120572 stimulatedmonocytic cells [42] These results indicated that the anti-malarial properties of pomegranate rindmay partly be due tourolithins

532 Histone Acetylation Status Histone acetylationdeacetylation status plays an important role in inflammationsince it is associated with the activationdeactivation oftranscription factors as NF-120581B and AP-1 directly implicatedin inflammation One of the mechanisms by which urolithinsexert their anti-inflammatory activity could be the inhibitionof histone acetyltransferases (HAT) as in fact has beendemonstrated for 5 120583M urolithin A and B that were able ofinhibiting HAT activity [53]

533 Anti-Inflammatory Effects on Human Colon FibroblastsColon fibroblasts have an important role in the gut immuneresponse and can be exposed to significant quantities ofcolon dietary metabolites Following ellagitannins intakeurolithin A and urolithin B as well as traces of ellagicacid can be found in the intestine Thus cultured humancolon fibroblasts stimulated with proinflammatory cytokineswere exposed to a mixture of urolithins A and B andellagic acid at concentrations representative of those thatmay be found in vivo (5ndash40120583M) [43] Urolithin A andmost significantly the mixture of metabolites were able toinhibit the migration capacity of the fibroblasts and theadhesion of monocytes to the fibroblasts giving evidenceof a potential amelioration of inflammation in the coloncells [43] Further molecular insights into these responsesrevealed that these effects were concomitant with a significantdownregulation of the levels of prostaglandin-E2 (PGE2)PAI-1 and interleukin 8 (IL-8) as well as of other keyregulators of cell migration and adhesion (Table 2)The studyof the individual metabolites indicated that urolithin A wasthe most active compound In addition to the inhibition ofPGE2 urolithin A and also urolithin B were able to inhibitthe expression of the two major enzymes responsible for thesynthesis of prostaglandins under inflammatory conditions(mPGES-1 and COX-2) whereas ellagic acid did not showany effect [44] The anti-inflammatory effects of urolithinsmay be mediated through regulation of the transcriptionfactor NF-120581B since both urolithins are able to inhibit theactivation of this factor and also exhibit an inhibitory effecton the mitogen-activated protein kinase pathways (MAPKpathways) c-Jun (urolithin A) and p38 (urolithins A and B)[44]


534 Anti-Inflammatory Effects on Human Endothelial CellsThe anti-inflammatory effects of urolithin glucuronides haverecently been investigated by Gimenez-Bastida et al [45]Using cytokine-induced human aortic endothelial cells thisstudy explored the effects of physiologically relevant con-centrations (low 120583M range 1ndash20120583M) of urolithin A andB glucuronides (the main metabolites detected in humanplasma after the oral intake of ellagitannin-containing foods[7]) on two early atherosclerotic events monocyte adhesionto endothelial cells and endothelial cell migration The mainoutcome of this research was that urolithin A glucuronidethe most abundant circulating conjugate after the intakeof ellagitannins exhibited the highest anti-inflammatoryactivity Urolithin A glucuronidemoderately but significantlyinhibited monocyte adhesion to the endothelial cells as wellas the migration capacity of these endothelial aortic cells[45] These effects were associated with a moderate butsignificant regulation of the levels of several key molecularmarkers associatedwith the atherosclerotic process (i) down-regulation of chemokine (C-C motif) ligand 2 (CCL2) (ii)downregulation of plasminogen activator inhibitor-1 (PAI-1)and (iii) regulation of several growth factors These resultswere in some cases comparable to those observed for thecorresponding aglycone urolithin A and suggested that theglucuronidation of urolithin A does not entirely eliminatethe activity of the aglycone and that the glucuronide maycontribute to the beneficial effects against cardiovasculardiseases attributed to the consumption of pomegranate (orpomegranate juice) [22] and other ellagitannin-containingfoods [2]

54 Anticarcinogenic-Related Activities

541 Topoisomerase II and CK2 Inhibitors The anticarcino-genic activity of urolithins is one of the most explored so farUrolithins seem to exert anticarcinogenic activity by affectingnumerous molecular pathways [5] Urolithins are inhibitorsof the CK2 enzyme a ubiquitous protein kinase implicatedin a wide variety of cell functions that when altered leadto processes like inflammation and cancer Using submi-cromolar concentrations in in silico screening urolithin Awas discovered to be a potent and selective CK2 inhibitor(IC50

= 039 120583M) [54] Urolithin M5 and another syntheticurolithin (never detected as an ellagic acid or ellagitanninmetabolite in animals) showed topoisomerase inhibitoryactivity at concentrations below 1 120583M showing an evenmore potent activity than that of the chemotherapeutic drugdoxorubicin The molecular mechanism of action towardhuman topoisomerase II seems to be the competition withATP for the ATP binding pocket of the human enzyme [55]

542 Human Prostate Cells The human prostate gland isone of the organs where urolithins can be detected afterthe consumption of pomegranate juice and walnuts [25] Inprostate cancer cells urolithins (A B C and D) inhibitedCYP1B1 activity (a target in prostate cancer chemopreven-tion) in a dose ranging from 115 120583M (urolithin A) to 137 120583M

(urolithin D) whereas higher concentrations were neededfor CYP1A1 activity inhibition (124ndash2907 120583M) The changesin 119881max and 119870

119898parameters with increasing concentrations

of CYP1B1 inhibitor suggested an uncompetitive inhibitionfor urolithin A The lack of changes for these parametershowever suggested a noncompetitive inhibition for urolithinB Furthermore the decrease in CYP1B1 activity exertedby urolithins was accompanied by a decrease in CYP1B1expression [46]

543 Anticancer Effects against Human Colon Cancer CellsThe in vitro studies conducted in colon cancer cells are ofgreat relevance since it is in this portion of the GI tract whereurolithins are produced and can reach bioactive concentra-tions A mixture of urolithin A urolithin B and ellagic acidat concentrations representative of those attainable in theintestine through the diet inhibited the proliferation of thehuman colon cancer Caco-2 cells [47] This inhibition wasmostly mediated through an S and G

2M cell cycle arrest in

association with the modulation of the expression of genesinvolved in cell cycle regulation (CCNB1 and CCNB1IP1)and in cancer development such as the oncogenes K-Rasand c-Myc the tumor suppressors DUSP6 and Fos and thegrowth factors receptors FGFR2 and EGFR [47]These resultsare concomitant with a general regulation of the ERK12signaling pathway Another pathway that may be a target forurolithins is the Wnt pathway Using a luciferase reporterof the canonical Wnt pathway in HEK T293 colon cellsSharma et al [48] showed that urolithin A was able toinhibit Wnt signaling with an IC

50of 39 120583M whereas ellagic

acid showed an IC50

of 63 120583M concentrations that can beachieved in the colon after pomegranate or ellagitannin-rich foods consumption [48] Kasimsetty et al [49] alsotested and compared the effects of urolithins on human coloncancer HT-29 cells and found that urolithins A B C andD were able to induce apoptosis in a concentration rangebetween 25 and 50120583M whereas much higher concentrations(sim500120583M) were necessary to promote cell cycle arrestUrolithins A B C and D also inhibited CYP1 activity (byapproximately 50) using concentrations in the range of50 to 75 120583M All these results suggest that regular dietaryconsumption of ellagitannin-containing foods which canyield and maintain 120583M concentrations of urolithins in thecolon may have a protective effect against colon cancerdevelopment

55 Protein Glycation Inhibitors Theproduction of advancedglycation end products is a secondary effect of hyperglycemiathat has a significant role in the cardiovascular complica-tions associated with diabetes and also with Alzheimerrsquosdisease Urolithins A and B (1120583M) showed significantantiglycative activity that increased when increasing theconcentrations up to 10120583M in the case of urolithin AThe same increase was not observed for urolithin B [40]This activity however was not related to their antioxidantactivity (measured as ABTS) or to their glyoxal-bindingcapacity


56 Antimicrobial Activity through the Inhibition of QuorumSensing Quorum Sensing (QS) is a bacterial communica-tion mechanism that responds to small molecules or auto-inducers and by which bacteria are able to detect popula-tion density regulate gene expression and control severalkey processes related to the infection progression such asvirulence biofilm formation andmotility An important areaof recent research focuses on finding natural compoundsable to inhibit QS that may pose an efficient alternative tothe use of antibiotics against pathogen infections Dietary-derived polyphenol metabolites present in the intestine maycontribute to reduce the ability of pathogens to invade theintestine through inhibition of their QS capacity UrolithinsA and B have been shown to reduce biofilm biomass andswimming motility of the enteropathogen Yersinia entero-colitica at concentrations as low as 4 120583M [56] These effectswere associated with a significant reduction of the levels ofthe bacteria autoinducers acylhomoserine lactones (AHLs)released by the bacteria to the culture media and wereaccompanied by the alteration of the expression levels ofgenes critically involved in the synthesis of lactones (yenIand yenR) and the synthesis of the flagella (flhDC fliAand fleB) [56] These results suggest that the microbiota-derived metabolites urolithin A and urolithin B may exertantipathogenic effects in the colon against Y enterocoliticaand may contribute to maintain the microbial equilibrium inthe gut

6 The In Vivo Evidence

The poor bioavailability of ellagitannins and ellagic acidderivatives as well as their extensive metabolism in thegastrointestinal tract has raised the question whether theseparent molecules found as such in the food were thereal active compounds systemically that could be relatedto the health benefits exerted by pomegranate juice andother ellagitannin-containing foodstuffs Our group reportedfor the first time the occurrence of urolithins in humansafter consuming pomegranate juice [7] These metabolitesreached micromolar concentrations in the bloodstreamand we launched the hypothesis that linked the potentialsystemic biological effects of pomegranate juice ingestionwith urolithins rather than with the polyphenols presentin pomegranate juice [7] or other ellagitannin-containingfoodstuffs such as walnuts strawberries and raspberries [8]

The first direct in vivo evidence regarding the biologicalactivity of urolithins was also reported by our group fiveyears later after launching the above hypothesis Larrosa etal [29] reported the anti-inflammatory and prebiotic effectsof the most significant urolithin metabolite (urolithin A)in a rat model of ulcerative colitis The rats received astandard chow supplemented eitherwith 250mgkgday of anellagitannin-rich pomegranate extract (PE) (HED sim25 g in a70 kg person) or with 15mgkgday of synthetic urolithin A(UroA) (HED sim154mg in a 70 kg person) for 25 days beforeinducing colon inflammation with dextran sodium sulfate(DSS)The pomegranate extract contained 35 punicalagins13 punicalin 45 ellagic acid glycosides and 89 free

ellagic acid Taking into account that urolithin A is the mainmetabolite produced after pomegranate intake the objectiveof that study was to evaluate a possible direct effect ofurolithin A ingested as synthetic compound and to comparethis effect with that exerted by the in vivo generated urolithinA after pomegranate extract consumption A number ofmeasurements were carried out including the evaluationof colon tissue damage microbiota changes antioxidantstatus prostaglandin E2 (PGE2) nitric oxide productioncyclooxygenase-2 (COX-2) inducible nitric oxide synthase(iNOS) prostaglandin E synthase (PTGES) gene expressionin colon mucosa (microarrays and RT-PCR) and polyphe-nol metabolism (LC-MS-MS) The administration of bothpomegranate extract and urolithin A for 25 days before theinduction of inflammation was reported to be safe accordingto serobiochemical analysis and evaluation of the animalsBoth pomegranate extract and urolithin-A decreased theanti-inflammatory markers iNOS COX-2 PTGES and PGE2in colonic mucosa However the anti-inflammatory activityexerted by 15mgkg urolithin A at both colonic and sys-temic levels was relatively stronger than that produced by250mgkg pomegranate extract Only urolithin A was ableto preserve the colonic architecture However pomegranateextract but not urolithin A decreased oxidative stress inplasma and colon mucosa (TBARs and FRAP methods)

Regarding gene expression analyses in the colon mucosaof a rat model of ulcerative colitis the expression of 2058and 6996 genes was found to be significantly modified bythe consumption of the pomegranate extract and urolithin Arespectively From those a total of 667 genes were commonlyregulated both by the pomegranate extract and urolithinA These genes were used for Functional Analyses usingthe Ingenuity Pathways Analysis (IPA Ingenuity SystemsRedwood City CA USA) Figure 5 shows the main topfunctions and diseases that were most significant to thegene expression altered by the intake of the pomegranateextract or urolithin A Of note some of the main functionsand disorders detected were associated to colon cancerdevelopment for example ldquocell deathrdquo ldquocell proliferationrdquoldquocancerrdquo ldquogastrointestinal diseaserdquo ldquoorganismal survivalrdquo andldquocell cyclerdquo

Using the Canonical Pathways Analysis tool of IPA itwas also possible to identify the main pathways from theIPA library that were most significant to the gene expressiondata analyzed Figure 6 shows themainmolecules specificallyinvolved in signaling pathways transcription regulation cellgrowth cell cycle and apoptosis for which deregulation hasbeen implicated in cancer development and whose transcriptlevels were significantlymodulated by urolithin A (upregu-lated in red downregulated in green)The differentmoleculesare depicted in their corresponding cellular compartmentand reported interactions between them are indicated byarrowsThis figure illustrates the complexity of the molecularmechanisms that may be triggered in a cell as a response tothe exposure to a potential bioactive compound for exampleurolithin A It should be noted that among the changesdetected some important tumor suppressors such as p53 andRb1 were upregulated whereas the antiapoptotic genes BclXL


0 5 10 15 20 25 30 35

Pomegranate extractUro-A

Cell deathCell proliferationProtein synthesis

CancerGastrointestinal disease

Hematological diseaseGenetic disorder

Respiratory diseaseOrganismal survival

Cell cycle

Threshold

minus log(119875-value)

Figure 5 Common top ten functions and diseases identified byfunctional analysis (IPA Ingenuity Pathways Analysis IngenuitySystems Redwood City CA USA) and that were most significantto gene expression altered by the intake of pomegranate extractor Urolithin A in the colon mucosa of a rat model of ulcerativecolitis (gene expression data from Larrosa et al [29]) Black barspomegranate extract Grey bars urolithin A A Fischerrsquos exact testwas used to calculate aP-value determining the probability that eachfunction or disease assigned to the data was due to chance We seta threshold at P value le 005 (dashed line) which corresponds to aFalse Discovery Rate (FDR) le 5 of false positives Note On the y-axis the significance is expressed as the minus log (10-based) of theP values The higher the bar the lower the P value and hence themore significant for the enrichment of the functiondisease

and Akt were downregulated by urolithin A These changeswere also observed after treatment with the pomegranateextract These results are indicative of in vivo molecularmodulation in the colonmucosa cells in response to urolithinA or pomegranate extract that may be associated with theprevention or reduction of malignant changes and cancerdevelopment In support of these results in vitro studies usinghuman colon cancer cells Caco-2 also reported the inhibitionof these cells proliferation and a cell cycle block by urolithinA(Table 2) In addition many of those genes involved in thesefunctions were found to be modulated by urolithin A in thehuman cells [47]

Our group also reported for the first time the in vivoprebiotic effect of pomegranate extract and urolithin A[29] Both pomegranate extract and urolithin A modulatedfavorably the gut microbiota in healthy rats (before inflam-mation) which could contribute to the protective effectsof pomegranate juice and urolithin A against the furtherinduced colon inflammation In the case of urolithin A theeffect was unequivocally due to synthetic metabolite orallyadministered and could be associated to the inhibition ofsome specific gut microbiota species to favor the growing ofsome (beneficial) microbial groups such as lactobacilli andbifidobacteria In the case of pomegranate extract the sameexplanation could be also given but a prebiotic effect exertedby other compounds present in the extract (fibers sugarsetc) could not be ruled out [29]

The second study dealing with the in vivo evidence ofurolithins was carried out by Ishimoto et al [57] Theseauthors reported the acute (24 h) anti-inflammatory effectof urolithin A on carrageenan-induced paw edema in miceIn this case urolithin A (300mgkg HED sim15 g in a 70 kgperson) was orally administered to mice at 1 or 6 h before theinjection of carrageenan The inflammatory effect was eval-uated for 24 h after carrageenan injection by measuring thehind paw volume Unfortunately no inflammatory markerswere measured in this study The authors claimed a rapidabsorption of urolithin A which peaked at 1 h in mice [57]However this was not in agreement with a previous studydealing with the pharmacokinetic and tissue distributionof urolithins where urolithin A peaked in plasma after 2 hbeing almost undetectable after 6 h [24] Ishimoto et al[57] described a high antioxidant activity of urolithin A inmouse plasma using the ORAC method and suggested thatthis activity could be correlated with the anti-inflammatoryeffects These results were not in agreement with those ofLarrosa et al [29] who suggested that anti-inflammatoryeffects of urolithin A were not mediated by its antioxidantactivity since no significant antioxidant effects in plasma andcolon of rats were found (TBARs and FRAP methods) Inaddition Cerda et al [7] did not observe any increase in theantioxidant status of human plasma after the intake of 1 L ofpomegranate juice for 5 days (ABTS and DPPH methods)Perhaps the high dose of urolithin A assayed by Ishimotoet al [57] as well as the specific use of the ORAC methodto measure the antioxidant activity could be behind thisapparent controversyOverall the human extrapolation of theresults obtained by Ishimoto et al [57] could be doubtful dueto their assay conditions These authors assayed a single doseof 15 g HED of urolithin A which raises safety concerns incontrast to the assay conditions approached by Larrosa et al[29]

7 Outlook and Further Research

Urolithins can reach significant concentrations in the humanbody after the intake of ellagitannin-containing productshowever the direct biological activity of urolithins has beenscarcely studied so far In this regard these metabolites couldbe themissing link to explain the health benefits associated tothe consumption of ellagitannin-containing products such aspomegranate There are a number of urolithins produced bythe human gut microbiota Some studies reveal the relevantturnover of these metabolites in the human body with a highurine excretion significant concentration in the bloodstreamand also disposition in the human prostate gland To dateonly two studies (animal models) have described the invivo anti-inflammatory [29 57] and possible cancer chemo-preventive activities [29] of the most abundant urolithinproduced in humans urolithin A In addition to this limitedevidence the direct in vivo activity of the rest of urolithins isnot yet known and thus more animal studies are needed Invitro studies with specific cell models and specificmetabolites(type of urolithin and conjugates such as glucuronides orsulfates) are useful to unravel mechanisms In this context


Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage

UV irradiation Cellproliferation

DNA repair

regulation

Repression ofapoptosis and

cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast

Proteasomaldegradation

NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2

Activation oftarget genes

Hedgehog target genes

WNT and PTCH regulation

Cyclin

Cyclin

regulation

Cell cycle arrest

Cell cycleregulation and

oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK

permeabilizationmembrane

Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300

CBPlowast CTNN TCF 34 SMAD4

SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu

120574 -Secretasecomplex

Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC

AXIN GSK3 NLKTAB1TAB2

TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP

Ras TGFTGF120573R2120573R1

TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B

Molecular mechanisms of cancer

c 2000-2010 Ingenuity systems Inc all rights reserved

Mitochondria

APH1PEN2

Figure 6 Graphical representation of main genes involved in signaling pathways transcription regulation cell growth cell cycle andapoptosis for which deregulation has been implicated in cancer development Genes depicted in red color were upregulated and genesrepresented in green color were downregulated in the colon mucosa of rats following the consumption of urolithin A (Larrosa et al [29])Analyses were carried out using the Canonical Pathways Analysis tool of Ingenuity Pathways Analysis (Ingenuity Systems Redwood CityCA USA)

appropriate concentrations and type of metabolites should becarefully chosen to draw relevant conclusions from these invitromodels

The evaluation of urolithins in humans could be moredifficult Although these metabolites are produced in thehuman gut the administration of synthetic urolithins tohumans can raise safety concerns since these metabolites arenot found as such (at least in relevant concentrations) in

foodstuffs and therefore they cannot be considered as dietarycompounds In this regard the toxicological evaluation ofurolithins in animal models is also missing

Conflict of Interests

The authors declare no conflict of interests


Acknowledgments

This work has been supported by the Projects Con-solider Ingenio 2010 CSD2007-00063 (Fun-C-Food) CICYTAGL2011-22447 Fundacion Seneca de la Region de Murcia(grupo de excelenciaGERM06 04486) andBACCHUS (FP7European Commission Grant Agreement 312090)

References

[1] M I Gil F A Tomas-Barberan B Hess-Pierce D M Holcroftand A A Kader ldquoAntioxidant activity of pomegranate juiceand its relationship with phenolic composition and processingrdquoJournal of Agricultural and Food Chemistry vol 48 no 10 pp4581ndash4589 2000

[2] M Larrosa M T Garcıa-Conesa J C Espın and F A Tomas-Barberan ldquoEllagitannins ellagic acid and vascular healthrdquoMolecular Aspects of Medicine vol 31 no 6 pp 513ndash539 2010

[3] F A Tomas-Barberan J C Espın and M T Garcıa-ConesaldquoBioavailability and metabolism of ellagic acid and ellagitan-ninsrdquo inChemistry and Biology of Ellagitannins S Quideau Edchapter 7 pp 293ndash297 World Scientific New Yersey NJ USA2009

[4] N P Seeram R N Schulman and D Heber PomegranatesAncient Roots to Modern Medicine CRC Press Boca Raton FLUSA 2006

[5] D Heber ldquoMultitargeted therapy of cancer by ellagitanninsrdquoCancer Letters vol 269 no 2 pp 262ndash268 2008

[6] B Cerda R Llorach J J Ceron J C Espın and F A Tomas-Barberan ldquoEvaluation of the bioavailability and metabolismin the rat of punicalagin and antioxidant polyphenol ofpomegranate juicerdquo European Journal of Nutrition vol 42 no1 pp 18ndash28 2003

[7] B Cerda J C Espın S Parra P Martınez and F A Tomas-Barberan ldquoThe potent in vitro antioxidant ellagitannins frompomegranate juice are metabolised into bioavailable but poorantioxidant hydroxy-6H-dibenzopyran-6-one derivatives bythe colonic microflora of healthy humansrdquo European Journal ofNutrition vol 43 no 4 pp 205ndash220 2004

[8] B Cerda F A Tomas-Barberan and J C Espın ldquoMetabolismof antioxidant and chemopreventive ellagitannins from straw-berries raspberries walnuts and oak-aged wine in humansidentification of biomarkers and individual variabilityrdquo Journalof Agricultural and Food Chemistry vol 53 no 2 pp 227ndash2352005

[9] N P Seeram SMHenning Y ZhangM Suchard Z Li andDHeber ldquoPomegranate juice ellagitannin metabolites are presentin human plasma and some persist in urine for up to 48 hoursrdquoJournal of Nutrition vol 136 no 10 pp 2481ndash2485 2006

[10] S U Mertens-Talcott P Jilma-Stohlawetz J Rios L Hingoraniand H Derendorf ldquoAbsorption metabolism and antioxidanteffects of pomegranate (Punica granatum L) polyphenols afteringestion of a standardized extract in healthy human volun-teersrdquo Journal of Agricultural and Food Chemistry vol 54 no23 pp 8956ndash8961 2006

[11] F A Tomas-Barberan N P Seeram and J C Espın ldquoBioavail-ability of pomegranate polyphenolsrdquo in Pomegranates AncientRoots to Modern Medicine N P Seeram R N Schulman andD Heber Eds Chapter 3 pp 45ndash60 CRC Press Boca RatonFL USA 2006

[12] B Cerda P Periago J C Espın and F A Tomas-BarberanldquoIdentification of urolithin A as a metabolite produced by

human colon microflora from ellagic acid and related com-poundsrdquo Journal of Agricultural and Food Chemistry vol 53 no14 pp 5571ndash5576 2005

[13] M A M Nawwar and A M A Souleman ldquo348910-Pentahydroxy-dibenzo[bd]pyran-6-one from Tamarix nilot-icardquo Phytochemistry vol 23 no 12 pp 2966ndash2967 1984

[14] M A M Nawwar S A M Hussein and I Merfort ldquoNMRspectral analysis of polyphenols from Punica granatumrdquo Phy-tochemistry vol 36 no 3 pp 793ndash798 1994

[15] E Lederer ldquoChemistry and biochemistry of some mammaliansecretions and excretionsrdquo Journal of the Chemical Society pp2115ndash2125 1949

[16] S J Jeong N Y Kim D H Kim et al ldquoHyaluronidaseinhibitory active 6H-dibenzo[bd]pyran-6-ones from the fecesof Trogopterus xanthipesrdquo Planta Medica vol 66 no 1 pp 76ndash77 2000

[17] S Ghosal ldquoChemistry of shilajit an immunomodulatoryAyurvedic rasayanrdquo Pure ampApplied Chemistry vol 62 no 7 pp1285ndash1288 1990

[18] G S Pope ldquoIsolation of two benzocoumarins from ldquocloverstonerdquo a type of renal calculus found in sheeprdquo BiochemicalJournal vol 93 no 3 pp 474ndash477 1964

[19] L J Filippich J Zhu P Oelrichs et al ldquoHepatotoxic andnephrotoxic principles in Terminalia oblongatardquo Research inVeterinary Science vol 50 no 2 pp 170ndash177 1991

[20] B Cerda J J Ceron F A Tomas-Barberan and J C EspınldquoRepeated oral administration of high doses of the pomegranateellagitannin punicalagin to rats for 37 days is not toxicrdquo Journalof Agricultural and FoodChemistry vol 51 no 11 pp 3493ndash35012003

[21] R Gonzalez-Barrio P Truchado H Ito J C Espın and FA Tomas-Barberan ldquoUV and MS identification of urolithinsand nasutins the bioavailable metabolites of ellagitannins andellagic acid in different mammalsrdquo Journal of Agricultural andFood Chemistry vol 59 no 4 pp 1152ndash1162 2011

[22] M Aviram L DornfeldM Rosenblat et al ldquoPomegranate juiceconsumption reduces oxidative stress atherogenic modifica-tions to LDL and platelet aggregation studies in humans andin atherosclerotic apolipoprotein E-deficient micerdquo AmericanJournal of Clinical Nutrition vol 71 no 5 pp 1062ndash1076 2000

[23] J C Espın R Gonzalez-Barrio B Cerda C Lopez-Bote AI Rey and F A Tomas-Barberan ldquoIberian pig as a model toclarify obscure points in the bioavailability and metabolismof ellagitannins in humansrdquo Journal of Agricultural and FoodChemistry vol 55 no 25 pp 10476ndash10485 2007

[24] N P Seeram W J Aronson Y Zhang et al ldquoPomegranateellagitannin-derivedmetabolites inhibit prostate cancer growthand localize to themouse prostate glandrdquo Journal of Agriculturaland Food Chemistry vol 55 no 19 pp 7732ndash7737 2007

[25] A Gonzalez-Sarrıas J A Gimenez-Bastida M T Garcıa-Conesa et al ldquoOccurrence of urolithins gut microbiota ellagicacid metabolites and proliferation markers expression responsein the human prostate gland upon consumption of walnuts andpomegranate juicerdquoMolecular Nutrition and Food Research vol54 no 3 pp 311ndash322 2010

[26] N P Seeram R Lee and D Heber ldquoBioavailability of ellagicacid in human plasma after consumption of ellagitannins frompomegranate (Punica granatum L) juicerdquoClinica Chimica Actavol 348 no 1-2 pp 63ndash68 2004

[27] B Doyle and L A Griffiths ldquoThe metabolism of ellagic acid inthe ratrdquo Xenobiotica vol 10 no 4 pp 247ndash256 1980


[28] M Azorın-Ortuno C Urban J J Ceron et al ldquoSafety evalu-ation of an oak-flavored milk powder containing ellagitanninsupon oral administration in the ratrdquo Journal of Agricultural andFood Chemistry vol 56 no 8 pp 2857ndash2865 2009

[29] M Larrosa A Gonzalez-Sarrıas M J Yanez-Gascon et alldquoAntiinflammatory properties of a pomegranate extract and itsmetabolite urolithin A in a colitis rat model and the effect ofcolon inflammation on the phenolic metabolismrdquo Journal ofNutritional Biochemistry vol 21 no 8 pp 717ndash725 2010

[30] H Ito ldquoMetabolites of the ellagitannin geraniin and theirantioxidant activitiesrdquo Planta Medica vol 77 no 11 pp 1110ndash1115 2011

[31] R Gonzalez-Barrio P Truchado R Garcıa-Villalba et alldquoMetabolism of oak leaf ellagitannins and urolithin productionin beef cattlerdquo Journal of Agricultural and Food Chemistry vol60 no 12 pp 3068ndash3077 2012

[32] B Cerda C Soto M D Albaladejo et al ldquoPomegranate juicesupplementation in chronic obstructive pulmonary diseasea 5-week randomized double-blind placebo-controlled trialrdquoEuropean Journal of Clinical Nutrition vol 60 no 2 pp 245ndash253 2006

[33] P TruchadoM LarrosaMTGarcıa-Conesa et al ldquoStrawberryprocessing does not affect the production and urinary excretionof urolithins ellagic acid metabolites in humans A cross-overstudyrdquo Journal of Agricultural and Food Chemistry vol 60 no23 pp 5749ndash5754 2012

[34] R Gonzalez-Barrio C A Edwards and A Crozier ldquoColonicCatabolism of ellagitannins ellagic acid and raspberry antho-cyanins in vivo and in vitro studiesrdquo Drug Metabolism andDisposition vol 39 no 9 pp 1680ndash1688 2011

[35] J J J van der Hooft R C H de vos V Mihaleva et al ldquoStruc-tural elucidation and quantification of phenolic conjugatespresent in human urine after tea intakerdquo Analytical Chemistryvol 84 no 16 pp 7263ndash7271 2012

[36] S Tulipani M Urpi-Sarda R Garcıa-Villalba et al ldquoUrolithinsare the main urinary microbial-derived phenolic metabolitesdiscriminating a moderate consumption of nuts in free-livingsubjects with diagnosed metabolic syndromerdquo Journal of Agri-cultural and Food Chemistry vol 60 no 36 pp 8930ndash89402012

[37] F Ferreres P Andrade M I Gil and F A Tomas-BarberanldquoFloral nectar phenolics as biochemical markers for the botan-ical origin of heather honeyrdquo Zeitschrift fur Lebensmittel-Untersuchung und-Forschung vol 202 no 1 pp 40ndash44 1996

[38] B P Moore ldquoCoumarin-like substances from Australian ter-mitesrdquo Nature vol 195 no 4846 pp 1101ndash1102 1962

[39] D Bioalonska S G Kasimsetty S I Khan and D FerreiraldquoUrolithins intestinal microbial metabolites of pomegranate simellagitannins exhibit potent antioxidant activity in a cell-basedassayrdquo Journal of Agricultural and Food Chemistry vol 57 no21 pp 10181ndash10186 2009

[40] E Verzelloni C Pellacani D Tagliazucchi et al ldquoAntiglyca-tive and neuroprotective activity of colon-derived polyphenolcatabolitesrdquo Molecular Nutrition and Food Research vol 55supplement 1 pp S35ndashS43 2011

[41] M Larrosa A Gonzalez-Sarrıas M T Garcıa-Conesa F ATomas-Barberan and J C Espın ldquoUrolithins ellagic acid-derived metabolites produced by human colonic microfloraexhibit estrogenic and antiestrogenic activitiesrdquo Journal ofAgricultural and Food Chemistry vol 54 no 5 pp 1611ndash16202006

[42] M DellrsquoAgli G V Galli M Bulgari et al ldquoEllagitannins of thefruit rind of pomegranate (Punica granatum) antagonize in vitrothe host inflammatory response mechanisms involved in theonset of malariardquoMalaria Journal vol 9 no 1 article 208 2010

[43] J A Gimenez-Bastida M Larrosa A Gonzalez-Sarrıas F ATomas-Barberan J C Espın andM T Garcıa-Conesa ldquoIntesti-nal ellagitannin metabolites urolithins and ellagic acid amelio-rate cytokine induced inflammation and associated molecularmarkers in human colon fibroblastsrdquo Journal of Agricultural andFood Chemistry vol 60 no 36 pp 8866ndash8876 2012

[44] A Gonzalez-Sarrıas M Larrosa F A Toms-Barberan PDolara and J C Espın ldquoNF-120581B-dependent anti-inflammatoryactivity of urolithins gut microbiota ellagic acid-derivedmetabolites in human colonic fibroblastsrdquo British Journal ofNutrition vol 104 no 4 pp 503ndash512 2010

[45] J A Gimenez-Bastida A Gonzalez-Sarrıas M Larrosa etal ldquoEllagitannin metabolites urolithin A glucuronide and itsaglycone urolithin A ameliorate TNF-120572 induced inflammationand associated molecular markers in human aortic endothelialcellsrdquo Molecular Nutrition amp Food Research vol 56 no 5 pp784ndash796 2012

[46] S G Kasimsetty D Bialonska M K Reddy C Thornton KL Willett and D Ferreira ldquoEffects of pomegranate chemicalconstituentsintestinal microbial metabolites on CYP1B1 in22Rv1 prostate cancer cellsrdquo Journal of Agricultural and FoodChemistry vol 57 no 22 pp 10636ndash10644 2009

[47] A Gonzalez-Sarrıas J C Espın F A Tomas-Barberan andM T Garcıa-Conesa ldquoGene expression cell cycle arrest andMAPK signalling regulation in Caco-2 cells exposed to ellagicacid and itsmetabolites urolithinsrdquoMolecular NutritionampFoodResearch vol 53 no 6 pp 686ndash698 2009

[48] M Sharma L Li J Celver C Killian A Kovoor and N PSeeram ldquoEffects of fruit ellagitannin extracts ellagic acid andtheir colonicmetabolite urolithin A onWnt signalingrdquo Journalof Agricultural and FoodChemistry vol 58 no 7 pp 3965ndash39692010

[49] S G Kasimsetty D Bialonska M K Reddy G Ma S I Khanand D Ferreira ldquoColon cancer chemopreventive activities ofpomegranate ellagitannins and Urolithinsrdquo Journal of Agricul-tural and Food Chemistry vol 58 no 4 pp 2180ndash2187 2010

[50] M V Selma J C Espın and F A Tomas-Barberan ldquoInteractionbetween phenolics and gut microbiota role in human healthrdquoJournal of Agricultural and Food Chemistry vol 57 no 15 pp6485ndash6501 2009

[51] F A Tomas-Barberan and C Andres-Lacueva ldquoPolyphenolsand health current state and progressrdquo Journal of Agriculturaland Food Chemistry vol 60 no 36 pp 8773ndash8775 2012

[52] H Ishimoto A Tai M Yoshimura et al ldquoAntioxidative prop-erties of functional polyphenols and their metabolites assessedby an ORAC assayrdquo Bioscience Biotechnology and Biochemistryvol 76 no 2 pp 395ndash399 2012

[53] A K Kiss S Granica M Stolarczyk and M F MelzigldquoEpigenetic modulation of mechanisms involved in inflamma-tion influence of selected polyphenolic substances on histoneacetylation staterdquo Food Chemistry vol 131 no 3 pp 1015ndash10202012

[54] G Cozza A Gianoncelli P Bonvini et al ldquoUrolithin as a con-verging scaffold linking ellagic acid and coumarin analoguesdesign of potent protein kinase CK2 inhibitorsrdquo ChemMed-Chem vol 6 no 12 pp 2273ndash2286 2011


[55] V Furlanetto G Zagotto R Pasquale S Moro and B GattoldquoEllagic acid and polyhydroxylated urolithins are potent cat-alytic inhibitors of human topoisomerase II an in vitro studyrdquoJournal of Agricultural and Food Chemistry vol 60 no 36 pp9162ndash9170 2012

[56] J A Gimenez-Bastida P Truchado M Larrosa et alldquoUrolithins ellagitannin metabolites produced by colon micro-biota inhibit quorum sensing in Yersinia enterocolitica phe-notypic response and associated molecular changesrdquo FoodChemistry vol 132 no 36 pp 1465ndash1474 2012

[57] H Ishimoto M Shibata Y Myojin et al ldquoIn vivo anti-inflammatory and antioxidant properties of ellagitanninmetabolite urolithin Ardquo Bioorganic amp Medicinal ChemistryLetters vol 21 no 19 pp 5901ndash5904 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


HO

HO

OO

OO

OO

OH

OH

Urolithin A

Coumarin Isocoumarin

Figure 1 Chemical structure of urolithin A a benzocoumarin

present in ellagic acid (lactonasedecarboxylase activity) andby successive removals of hydroxyls (dehydroxylase activi-ties) [12] (Figure 2)

Urolithins are not commonmolecules in nature but theyhave been reported in plants rich in ellagitannins as it is thecase of Tamarix nilotica flowers [13] and Punica granatumleaves that contain urolithin M-5 (348910-pentahydroxy-dibenzo[bd]pyran-6-one) [14] They have also been foundin some herbivore-derived products such as the castoreumproduced by beavers [15] a product of interest for theperfume industry and thePteropi faeces (faeces of the squirrelTrogopterus xanthipes) that are used in traditional Chinesemedicine [16] Urolithins are also relevant constituents ofshilajit a soil-derived medicinal product used in Ayurvedicmedicine [17]

The name urolithin was first given to two metabolitesisolated from the renal calculus of sheep (Trifolium sub-terraneum has been reported as the cause of clover stoneand might be a relevant source of ellagitannins) that werenamed urolithin A and urolithin B [18] These two moleculescoincided with pigments I and II previously described fromcastoreum and beaver glands [15]

The intake of large amounts of ellagitannins and particu-larly of punicalagin from Terminalia oblongata by cattle wasassociated with hepatotoxicity and nephrotoxicity [19] How-ever the subchronic oral administration of large amountsof pomegranate ellagitannins was not toxic to rats wherethese compounds were found to be extensively metabolizedto urolithins [20]

Since urolithins are ellagitannin-derived catabolites thatcan be absorbed and reach different tissues in the body theyhave been suggested as the molecules potentially respon-sible for the biological effects observed as a consequenceof the consumption of pomegranate or other ellagitannin-containing foods [3 7 9 11] Here we review the state of theart in urolithin metabolites production by gut microbiotatheir absorption tissue distribution and pharmacokineticsthe cell andmolecular mechanisms for their biological effectsreported so far using different in vitromodels and the in vivoevidence in animals and humans

2 Urolithins Chemical NatureDetection and Identification

Urolithins constitute a whole metabolic family producedby the opening and decarboxylation of one of the lactonerings of ellagic acid and the sequential removal of hydroxylsfrom different positions (Figure 2) After decarboxylation thefirst metabolite is urolithin M-5 (pentahydroxy-urolithin)and from this several tetrahydroxy-urolithin isomers areproduced by removal of one hydroxyl group from differentpositions (urolithin D urolithinM-6) Trihydroxy-urolithins(urolithin C urolithin M-7) were then produced after theremoval of a second hydroxyl and dihydrox- urolithins(urolithin A and isourolithin A) after the removal of athird one Monohydroxy-urolithin (urolithin B) was alsodetected particularly in those cases in which isourolithin Awas produced (Figure 2) Further degradation of urolithinsto remove the second lactone ring has not been reportedso far although it should not be discarded Urolithins showcharacteristic UV spectra that can be used for the identi-fication of different hydroxyl-substitution patterns on theurolithin nucleus as well as the further conjugation withmethyl glucuronic acid or sulfate [21] The UV spectra ofurolithins in methanol exhibit two major absorption bandsin the region of 240ndash400 nm (Figure 4) These are referred toas band I (300ndash380 nm) and band II (240ndash280 nm) In manycases an additional band III (between 280 and 300 nm) is alsoobserved Unfortunately bands I and II cannot be associatedwith a specific ring of the urolithin nucleus [21] Two groupsof urolithinUV spectra were distinguished after a study of theplots of the different spectra urolithins with a hydroxyl in the9-position and those without hydroxylation in the 9-position(Figure 4) The occurrence of the hydroxyl at position 9 as isthe case of isourolithin A produces a hypsochromic shift inband I and an increase in the absorption of band II displayinga maximum around 256 nm which is the main absorptionband of the spectrum of urolithins with a hydroxyl in the 9-position Conjugation with glucuronic acid or sulfate also hasmeasurable effects on the UV spectrum and could be used asa diagnostic method for urolithin metabolites identification

3 Bioavailability Metabolism and TissueDistribution of Pomegranate Ellagitanninsand Their Metabolites

Due to the potential health effects and the expectationraised by the high antioxidant activity of pomegranate juicesthat was thought to be due to punicalagins punicalins andother ellagitannins [1 22] a sensible step in the researchwas the determination of the absorption and metabolism ofthese compounds in animals and humans The first studytesting the bioavailability of pomegranate ellagitannins wascarried out on rats and this showed that after the intake oflarge amounts of pomegranate husk ellagitannins the mainmetabolites detected in plasma and urine were urolithinsA B and C (Figure 2) and smaller amounts of ellagicacid-dimethyl ether glucuronide [6] Interestingly a smallamount of punicalagin was also detected in plasma and urine


O

OO

O

OO

COOH

OO

OO

OO

OO O

O

OO

OO

OOO

O

Ellagitannins

Ellagic acid

Urolithin M-5

Urolithin D


Urolithin A

Urolithin B

Intermediate

Urolithin M-6

Isourolithin A

Isourolithin B

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HOHO

HO

HO

HO

HO

HO

OH

OHOH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OHOH

OH







OO

Urolithin A(URO-A)

OO

Ellagitannins

Ellagic acid

O

OO

O

OO

Urolithin B(URO-B)

OO

Esophagus



HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

OH

OH

Bact

eria

l met

abol

ism







OO

246

200 250 300 350 400

mAU

0

100

200

300 280

305

356


120582 (nm)

Urolithin A

HO OH

(a)

OO

256

200 250 300 350 400

mAU

0

500

1000

1500

2000

2500

302329



HO

OH

(b)



















[7 9 2532]




















[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















O

OO

O

OO

COOH

OO

OO

OO

OO O

O

OO

OO

OOO

O

Ellagitannins

Ellagic acid

Urolithin M-5

Urolithin D


Urolithin A

Urolithin B

Intermediate

Urolithin M-6

Isourolithin A

Isourolithin B

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HO

HOHO

HO

HO

HO

HO

HO

OH

OHOH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OH

OHOH

OH







OO

Urolithin A(URO-A)

OO

Ellagitannins

Ellagic acid

O

OO

O

OO

Urolithin B(URO-B)

OO

Esophagus



HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

OH

OH

Bact

eria

l met

abol

ism







OO

246

200 250 300 350 400

mAU

0

100

200

300 280

305

356


120582 (nm)

Urolithin A

HO OH

(a)

OO

256

200 250 300 350 400

mAU

0

500

1000

1500

2000

2500

302329



HO

OH

(b)



















[7 9 2532]




















[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















OO

Urolithin A(URO-A)

OO

Ellagitannins

Ellagic acid

O

OO

O

OO

Urolithin B(URO-B)

OO

Esophagus



HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

OH

OH

Bact

eria

l met

abol

ism







OO

246

200 250 300 350 400

mAU

0

100

200

300 280

305

356


120582 (nm)

Urolithin A

HO OH

(a)

OO

256

200 250 300 350 400

mAU

0

500

1000

1500

2000

2500

302329



HO

OH

(b)



















[7 9 2532]




















[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















OO

246

200 250 300 350 400

mAU

0

100

200

300 280

305

356


120582 (nm)

Urolithin A

HO OH

(a)

OO

256

200 250 300 350 400

mAU

0

500

1000

1500

2000

2500

302329



HO

OH

(b)



















[7 9 2532]




















[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


























[7 9 2532]




















[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
























[41]


of MMP-9 [42]



[43]




















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















50=





















acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





























acid showed an IC50












0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

























0 5 10 15 20 25 30 35






Cell cycle

Threshold









Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Plasma membrane

PI3KAKTSignaling

JNK

Ras

Ras

Ras

RasGEF

Ras

DAG

IP3

PIP2

PIP2PIP3

Ca2+

PKC

Ras

CAMKII

GRP

GRFRas

GAPlowast SynGAP

MP1

MP1

ERK

ERK12

MEK12

MEK12

c-Raf

AKT

PI3K

PKA

G120572

PLC AC C3G

ATP

EPAC1

cAMP Rap

Rho

B-Raf

CRK

RTKGPCR

Hormones

Src

SOS

SHCGRB2

SOS

SHCGRB2 SOS

SHC

SHP2GRB2Cdc42

FAK

JAK

PI3K PI3K

Integrin

ECMprotein

Cytokines

Cytokinereceptor

120573

RaIGEFRaf

Fyn

AB

RalGAP

RAC

PAK

c-Raf

GSK3 I120581B

I120581B

NF-120581B MDM2

NF1lowast

AKT

PI3K

GAB12 CbI IRS1

RTK

Growthfactors

Growthfactors

Cyclin

NucleusDNA damage


DNA repair

regulation


cell cyclearrest

Cellsurvivalc-Jun

c-Fos

c-Jun

c-FosBIM

FoxO1

Noxa

BAD

tBID

BclXL

PUMAMMP

Bcl2

Elk-1

JNK

JNK

12

ERK12

ERK12

P53lowast

P53lowast


NF-120581BFANCD2

NBS1

ATRATMDNA

PK

Chk1Chk2

CyclinD

AuroraA

HIPK2




Cyclin

Cyclin

regulation

Cell cycle arrest


oncogenesis

Cell survival

NICD

Gli

MMP mitochondrial

CyclinsCDKsCKIs

c-Jun

c-Fos

c-Fos

CytoC

BAX

BAK


Apoptosis

SMAC

JNK

p38MAPK

MKK36

CIAP

BRCA1

p16INK4AP

Rb

Rb

E2FE2F

DC25

CDK46

D

CyclinE

p19

p27

p21

p21

p14ARF

p18

MAX

Myc

MIZ1

INK4DINK4C p15

INK4B

CDK2+

120573

HIF1120572

120572

NICD

RBPJK HAT1

NICD P300


SMAD4

SMAD158

SMAD23E2F

RBL1

KIP1

LEF1

Caspase

Caspase

9

MDM2

CIP1

CIP1

CytoC

APAF1

NICD

Gli

SuFuFu


Dsh

BID tBID

FLIP

Fas

FasL5A

WNT

WNT

PTCH Smo

1

JNK

TAK1

TAB2 TAB1

ERK12protein

Hedgehog

G

E

120573

120573

CTNN

120573CTNN

CTNN CTNN

120573CTNN SMAD4

SMAD4

SMAD6

SMAD7

SMAD7

SMAD4SMAD

SMAD

158

SMAD158

158

23

SMAD23

APC


TAK1

PSEN12

NCSTN

Notch1 Fzd LRP CDH

Src120575

BMPR1 BMPR2

BMP


TGF-120573

FADDDaxx

ASK1

Caspase810

367

Cd2+

NF-120581B



Mitochondria

APH1PEN2








Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Acknowledgments


References


































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















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